针刀松解法对膝骨关节炎兔软骨基质Col-Ⅱ、Aggrecan基因及蛋白表达的影响
摘要
骨关节炎是一种退行性骨关节病,是中老年常见病、多发病,在最常见的三大老年病(心脑血管疾病、骨质疏松症、骨性关节炎)中,骨性关节炎发病率在世界范围内居于首位。骨关节炎好发于负重较大的脊柱,膝、髋关节等部位,主要病变是关节软骨的退行性变和继发性骨质增生,临床表现为缓慢进行性发展的关节疼痛、肿胀、僵硬、伴活动受限。严重的影响了患者日常生活和工作,甚至可造成残疾。随着人口老龄化进程加速,这一问题日益突出。因此骨关节炎的防治工作已受到社会广泛关注,对骨关节炎进行研究有着重要的现实意义。目前该病的病因及发病机制还不太清楚。
由于膝关节负重量与活动量较大,因此膝骨关节炎(KOA)也是骨关节炎最常见的一种,一般保守治疗较难且易复发。30多年来,针刀作为一种中医新的微创技术,在KOA的治疗方面取得了较好的疗效,但基础研究尚待深入。本研究通过实验来探讨针刀治疗膝骨关节炎的可能机制,为以后研究奠定基础,也为临床治疗提供理论依据。
研究目的
本实验采用针刀干预并与电针组对照,观察针刀干预对骨关节炎动物模型的关节软骨组织中II型胶原(Col-Ⅱ)、聚集蛋白聚糖(Aggrecan)、基质金属蛋白酶-3(MMP-3)基因、蛋白表达的影响,从分子生物学角度探讨针刀干预膝骨关节炎的可能机制,为以后研究奠定基础,也为临床治疗膝骨关节炎提供理论依据。
实验方法
将6月龄健康新西兰兔(雌雄各半)随机分为空白组、模型组、针刀组和电针组,采用左后肢伸直位固定制动法(Vidman法)制备膝骨关节炎动物模型。针刀组及电针组于造模结束1周后开始进行干预,针刀组每周干预1次,共干预三周,电针组隔天治疗1次,1周治疗3次,共治疗3周。治疗前后分别观察兔膝关节的行为学指标；大体观察各组关节软骨和滑膜的病理表现；光镜观察各组兔软骨切片的病理表现；Real-timePCR法检测Col-ⅡmRNA、 AggrecanmRNA、 MMP-3mRNA的表达水平,Western blot法检测Col-Ⅱ、Aggrecan、 MMP-3蛋白的表达水平。
结果
1大体观察：空白组兔膝关节无肿胀、充血,关节液透明,关节软骨呈淡蓝色、半透明、光滑,滑膜无水肿、肥厚；模型组兔膝关节明显肿胀、充血,关节液呈浅黄色稍混浊,关节软骨呈灰白色、无光泽,不光滑、不透明,有缺损,可见明显溃疡面及裂隙,有的可见软骨下骨裸露,胫骨内侧髁较明显,滑膜明显充血,水肿、肥厚；针刀组兔左膝关节轻度肿胀无明显充血,软骨颜色淡白,色泽较亮,有轻度缺损,未见到明显溃疡面,关节液透明稍浅黄,与模型组相比水肿与充血程度明显减轻；电针组关节软骨透明度减低欠光泽,有的可见裂隙,滑膜增生减轻,未见明显充血。说明针刀、电针治疗具有减轻软骨损伤、关节水肿的作用。
2光镜观察：空白组兔膝关节软骨结构完整,关节软骨表面光滑,厚度正常,软骨细胞排列整齐,分层清楚,排列有序,软骨基质均匀,潮线较清晰完整,无血管翳生成。模型组软骨表面不光滑,表层软骨细胞稀少,细胞排列明显不规则,层次紊乱,有时可见剥脱缺损深达潮线,软骨细胞巢居,潮线模糊或见双重潮线。针刀组与电针组软骨细胞排列整齐,少见软骨裂隙,但细胞排列仍见局部紊乱,偶见重复潮线,软骨表层比较光滑,未见血管翳形成。对切片按照Mankin法进行评分,四组评分总体差别极显著(P<0.01)。针刀组与模型组相比,具有极显著差异(P<0.01),与空白组比较差异显著(P<0.05),与电针组相比无显著差异(P>0.05)。电针组与空白组相比有显著差异(P<0.01),与模型组相比差异不显著(P>0.05)。针刀松解法有保护软骨的作用。
3行为学改变：模型组、针刀组和电针组Lequesne MG评分分值治疗前统计无显著性差异(P>0.05),治疗后和治疗前后计分差值有显著性差异(P<0.05)。针刀松解法有效地改善了兔KOA关节疼痛、活动度、肿胀和步态。
4Real-time PCR实验结果显示各组Col-II mRNA的含量表达针刀组较模型组明显升高,二者有显著差异(P<0.05)；电针组较模型组明显升高,二者有显著差异(P<0.05)；电针组与空白组有明显差异(P<0.05)；各组软骨AggrecanmRNA含量表达模型组较空白组明显降低,二者相比有极显著差异(P<0.01)；MMP-3mRNA表达模型组较空白组明显升高,二者相比有显著差异(P<0.05)；针刀组较模型组降低,但与空白组相比无明显差异(P>0.05),针刀组较模型组降低,但二者无显著差异(P>0.05)。说明针刀松解能有效上调Col-ⅡmRNA和AggrecanmRNA表达,下调MMP-3mRNA表达,针刀组效果优于电针组。
5Western blot实验结果显示各组软骨Co1-Ⅱ蛋白含量表达模型组较空白组明显降低,二者相比有极显著差异(P<0.01),针刀组较模型组明显升高,二者有有极显著差异(P<0.01),针刀组与电针组比较有显著差异(P<0.05)；Aggrecan蛋白含量表达针刀组较模型组明显升高,二者有显著差异(P<0.05),电针组与模型组无明显差异(P>0.05)。MMP-3蛋白表达模型组较空白组明显升高,二者相比有极显著差异(P<0.01),针刀组较模型组明显降低,二者有有极显著差异P<0.01),针刀组与电针组比较有显著差异(P<0.05)。说明针刀松解能有效上调Co1-Ⅱ蛋白和Aggrecan蛋白表达,下调MMP-3蛋白表达,针刀组效果优于电针组。
结论
1模型组兔膝关节软骨MMP-3的基因及蛋白含量表达明显升高,而软骨基质Co1-Ⅱ和Aggrecan的基因及蛋白含量表达明显降低,说明基质金属蛋白酶能很好反映膝关节软骨代谢变化,它对关节软骨基质具有降解作用,能加速软骨退变。
2针刀松解法能使关节软骨中MMP-3的基因及蛋白含量表达降低,基质中的Col-Ⅱ和Aggrecan的基因及蛋白含量表达升高,说明针刀松解法有一定的抑制细胞外基质胶原降解,保护促进和刺激软骨修复的作用,为针刀松解法治疗膝骨性关节炎提供了实验研究的依据,揭示了针刀松解法治疗膝关节骨性关节炎的科学内涵。
Osteoarthritis is a common degenerative joint disease, more common and frequently in the elderly, in three of the most common diseases by elderly patients (the most common cardiovascular disease, osteoporosis, osteoarthritis), the incidence rate of osteoarthritis in the first place in the world.Osteoarthritis occurs in the weight-bearing larger spine, knee, hip and other parts, and the main lesion is degeneration of articular cartilage and secondary bone hyperplasia, clinical manifestations of slowly progressive development of joint pain, swelling, stiffness, with limited mobility.Severely affected patients in daily life and work, and even can cause disability. With the population aging process accelerated, this problem becomes increasingly prominent. Therefore, the prevention and treatment of osteoarthritis has received widespread attention in the community, and the osteoarthritis research has important practical significance.Currently the etiology and pathogenesis of this disease is not clear.
Due to weight bearing of knee joint and activity of it is larger, so the knee osteoarthritis (KOA) is one of the most common bone arthritis, generally conservative treatment is more difficult and easy recurrence.For more than30years, acupotomy as a new, minimally invasive technique of traditional Chinese medicine,had better curative effect in the treatment of KOA, but basic research remains to be in-depth.This study through experiments to explore possible mechanisms of acupotomy for treating knee osteoarthritis, lay a foundation for further research, and provide theoretical basis for clinical treatment.
Objective
The experiment used acupotomy intervention and compared with electric acupuncture group, to observe acupotomy intervention on animal models of osteoarthritis articular cartilage tissue type Ⅱcollagen (Col-Ⅱ), aggrecan (Aggrecan), matrix metalloproteinases-3(MMP-3) gene and protein expression, to explore the possible mechanism of knee osteoarthritis acupotomy intervention from the perspective of molecular biology, to lay the foundation for the later research, and also provide a theoretical basis for clinical treatment of knee osteoarthritis.
Methods
Six-month-old healthy adult New Zealand rabbits(equal between genders) were divided into four groups randomly:normal control group, model group, acupotomy group,electroacupunctur(EA) group. The left hind limb immobilization in extension position braking method (Vidman method) for preparing animal models of knee osteoarthritis. Acupotomy group and electro-acupuncture group began to intervene after one week at the end of molding, acupotomy group intervention for one time per week, a total of three weeks of intervention, electric acupuncture group was treated one time on alternate days, treatment three times in one weeks, a total of three weeks of treatment. Before and after treatment knee joint behavior index were observed in rabbit; gross observed pathological manifestations of the articular cartilage and synovial membrane; pathology of rabbit cartilage slices were observed under optical microscope; Real-time PCR method to detect Col-ⅡmRNA, AggrecanmRNA, MMP-3mRNA expression level, and Western blot method to detect Col-Ⅱ,Aggrecan, MMP-3protein expression level.
Results
1General observation:Normal control group rabbits knee joints were not swelling, congestion, joint fluids transparent, articular cartilages pale blue, translucent, smooth. Synovial membranes was without edema, hypertrophy.Model group rabbits knee joints were significantly swelling or congestion. Joint fluid was pale yellow and slightly turbid.Articular cartilages were gray, dull, not smooth, and not opaque.There were defect.Ulcers and fissures could be seen clearly. Some subchondral bone was exposed, medial tibial condyle more obvious, synovial hyperemia, edema, hypertrophy.Acupotomy group rabbits the left knee joints mild swelling and had no significant congestion, cartilage pale color, bright color, mild defect, and the obvious ulcers was not seen.Joint fluid was transparent and slightly yellow.Compared with the model group,edema and hyperemia was significantly ease. EA group articular cartilages transparency reduced and were less shin. Some visible cracks could be seen.Synovial hyperplasia reduced,and there was not significant congestion.Indicating acupotomy, electro-acupuncture treatment can relieve cartilage damage,and joint edema.
2Optical microscope observation:Knee joint articular cartilages of normal control group were structural integrity.Articular cartilage surface was smooth.Thickness was normal. Cartilage cells arranged in neat rows, and layered clear, orderly. Cartilage matrix was homogencously,and tide line was clear and complete, no pannus formation. In model group, the cartilage surface was not smooth.The surface of cartilage cells were scarce.Cells arranged irregularly. Levels were disturbance.Sometimes visible peeling defect was as deep as the tide line. Chondrocytes nested.Tide line blurred or double tide line was seen, part of the cartilage pannus formation and vascular. Acupotomy group and electro-acupuncture group cartilage cells arranged in neat rows. Cartilage fracture was rare.But local cells arranging in disorder still can be seen. Repetitive tide line was occasionally seen.Cartilage surface was smooth,without pannus formation. The sections were scored according to Mankin method.Four groups overall score were significantly different (P<0.01). Acupotomy group comparing with model group had significant difference (P<0.01); and compareing with the control group had significant difference (P<0.05).There was not significant difference as compared with electroacupuncture group (P>0.05). EA group comparing with the control group had significant difference (P<0.01), and no significant difference as compared with the model group (P>0.05). Acupotomy method had cartilage protective effect.
3Behavior changes:the model group and acupotomy group, acupuncture group Lequesne MG score before treatment had no significant statistical difference (P>0.05).The score difference before and after treatment, and score after treatment had significant difference (P<0.05).Acupotomy therapy is effective in improving the rabbit KOA joint pain, swelling and gait, activity.
4Real-time PCR experimental results showed Col-ⅡmRNA content expression of acupotomy group increased more significantly than that of model group, and there were significant differences between the two (P<0.05). Compared with model group, Col-ⅡmRNA content expression of electro-acupuncture group significantly increased, and there were significant differences between them(P<0.05). Electroacupuncture group had significant difference with normal control group (P<0.05). Cartilage AggrecanmRNA content expression of model group was significantly decreased,compared with control group,and there were significant differences between the two compared (P<0.01).The expression of MMP-3mRNA in model group was significantly increased compared with normal control group, and there were significant differences between the two (P<0.05).Each group decreased as compared with the model group, but no significant difference compared with the normal control group (P>0.05).Acupotomy group decreased as compared with the model group, but no significant difference between the two (P>0.05). The results of Real-time PCR tests showed that acupotomy could effective up-regulated expression of Col-ⅡmRNA and AggrecanmRNA, down-regulated MMP-3mRNA expression, and the effect of acupotomy group was better than electroacupuncture group.
5Western blot experimental results showed Coll-Ⅱprotein expression in cartilage of model group significantly decreased than that of normal control group group, and there were significant differences between the two (P<0.01). Acupotomy group increased significantly compared with model group, the two had very significant difference (P<0.01). There was significant difference between acupotomy group and electroacupuncture group (P<0.05).The expression of aggrecan protein in acupotomy group was higher than that of model group, and there was significant differences between the two (P<0.05. Electroacupuncture group and model group had no significant difference (P>0.05). The expression of MMP-3protein in model group was significantly increased compared with normal control group, and there was significant differences between the two(P<0.01), Acupotomy group was significantly lower than the model group, and the two had significant differences(P<0.01).There was significant difference between acupotomy group and the electroacupuncture group (P<0.05). The results of Real-time PCR tests showed that acupotomy could effectively increase expression of Col-Ⅱprotein and Aggrecan protein,reduced expression of MMP-3protein, and the effect of acupotomy group was better than control group.
Conclusions
1Gene and protein expression of MMP-3in articular cartilage model group rabbits significantly increased, while the gene and protein content expression in the cartilage matrix of Coll-Ⅱand Aggrecan decreased obviously, sand howed that matrix metalloproteinases could well reflect the changes of knee cartilage metabolism, It had a role in the degradation of the extracellular matrix of articular cartilage, and could accelerate the degeneration of cartilage.
2Acupotomy can make the gene and protein content expression of MMP-3in articular cartilage decreased, and gene and protein content expression in the matrix of Col-Ⅱand Aggrecan elevated, showing that acupotomy was inhibition of extracellular matrix collagen degradation, protection to promote and stimulate cartilage repair function.Provided the basis for the experimental study of acupotomy treatment of osteoarthritis of the knee, and reveaed the scientific connotation of acupotomy in treating knee joint osteoarthritis.
由于膝关节负重量与活动量较大,因此膝骨关节炎(KOA)也是骨关节炎最常见的一种,一般保守治疗较难且易复发。30多年来,针刀作为一种中医新的微创技术,在KOA的治疗方面取得了较好的疗效,但基础研究尚待深入。本研究通过实验来探讨针刀治疗膝骨关节炎的可能机制,为以后研究奠定基础,也为临床治疗提供理论依据。
研究目的
本实验采用针刀干预并与电针组对照,观察针刀干预对骨关节炎动物模型的关节软骨组织中II型胶原(Col-Ⅱ)、聚集蛋白聚糖(Aggrecan)、基质金属蛋白酶-3(MMP-3)基因、蛋白表达的影响,从分子生物学角度探讨针刀干预膝骨关节炎的可能机制,为以后研究奠定基础,也为临床治疗膝骨关节炎提供理论依据。
实验方法
将6月龄健康新西兰兔(雌雄各半)随机分为空白组、模型组、针刀组和电针组,采用左后肢伸直位固定制动法(Vidman法)制备膝骨关节炎动物模型。针刀组及电针组于造模结束1周后开始进行干预,针刀组每周干预1次,共干预三周,电针组隔天治疗1次,1周治疗3次,共治疗3周。治疗前后分别观察兔膝关节的行为学指标；大体观察各组关节软骨和滑膜的病理表现；光镜观察各组兔软骨切片的病理表现；Real-timePCR法检测Col-ⅡmRNA、 AggrecanmRNA、 MMP-3mRNA的表达水平,Western blot法检测Col-Ⅱ、Aggrecan、 MMP-3蛋白的表达水平。
结果
1大体观察：空白组兔膝关节无肿胀、充血,关节液透明,关节软骨呈淡蓝色、半透明、光滑,滑膜无水肿、肥厚；模型组兔膝关节明显肿胀、充血,关节液呈浅黄色稍混浊,关节软骨呈灰白色、无光泽,不光滑、不透明,有缺损,可见明显溃疡面及裂隙,有的可见软骨下骨裸露,胫骨内侧髁较明显,滑膜明显充血,水肿、肥厚；针刀组兔左膝关节轻度肿胀无明显充血,软骨颜色淡白,色泽较亮,有轻度缺损,未见到明显溃疡面,关节液透明稍浅黄,与模型组相比水肿与充血程度明显减轻；电针组关节软骨透明度减低欠光泽,有的可见裂隙,滑膜增生减轻,未见明显充血。说明针刀、电针治疗具有减轻软骨损伤、关节水肿的作用。
2光镜观察：空白组兔膝关节软骨结构完整,关节软骨表面光滑,厚度正常,软骨细胞排列整齐,分层清楚,排列有序,软骨基质均匀,潮线较清晰完整,无血管翳生成。模型组软骨表面不光滑,表层软骨细胞稀少,细胞排列明显不规则,层次紊乱,有时可见剥脱缺损深达潮线,软骨细胞巢居,潮线模糊或见双重潮线。针刀组与电针组软骨细胞排列整齐,少见软骨裂隙,但细胞排列仍见局部紊乱,偶见重复潮线,软骨表层比较光滑,未见血管翳形成。对切片按照Mankin法进行评分,四组评分总体差别极显著(P<0.01)。针刀组与模型组相比,具有极显著差异(P<0.01),与空白组比较差异显著(P<0.05),与电针组相比无显著差异(P>0.05)。电针组与空白组相比有显著差异(P<0.01),与模型组相比差异不显著(P>0.05)。针刀松解法有保护软骨的作用。
3行为学改变：模型组、针刀组和电针组Lequesne MG评分分值治疗前统计无显著性差异(P>0.05),治疗后和治疗前后计分差值有显著性差异(P<0.05)。针刀松解法有效地改善了兔KOA关节疼痛、活动度、肿胀和步态。
4Real-time PCR实验结果显示各组Col-II mRNA的含量表达针刀组较模型组明显升高,二者有显著差异(P<0.05)；电针组较模型组明显升高,二者有显著差异(P<0.05)；电针组与空白组有明显差异(P<0.05)；各组软骨AggrecanmRNA含量表达模型组较空白组明显降低,二者相比有极显著差异(P<0.01)；MMP-3mRNA表达模型组较空白组明显升高,二者相比有显著差异(P<0.05)；针刀组较模型组降低,但与空白组相比无明显差异(P>0.05),针刀组较模型组降低,但二者无显著差异(P>0.05)。说明针刀松解能有效上调Col-ⅡmRNA和AggrecanmRNA表达,下调MMP-3mRNA表达,针刀组效果优于电针组。
5Western blot实验结果显示各组软骨Co1-Ⅱ蛋白含量表达模型组较空白组明显降低,二者相比有极显著差异(P<0.01),针刀组较模型组明显升高,二者有有极显著差异(P<0.01),针刀组与电针组比较有显著差异(P<0.05)；Aggrecan蛋白含量表达针刀组较模型组明显升高,二者有显著差异(P<0.05),电针组与模型组无明显差异(P>0.05)。MMP-3蛋白表达模型组较空白组明显升高,二者相比有极显著差异(P<0.01),针刀组较模型组明显降低,二者有有极显著差异P<0.01),针刀组与电针组比较有显著差异(P<0.05)。说明针刀松解能有效上调Co1-Ⅱ蛋白和Aggrecan蛋白表达,下调MMP-3蛋白表达,针刀组效果优于电针组。
结论
1模型组兔膝关节软骨MMP-3的基因及蛋白含量表达明显升高,而软骨基质Co1-Ⅱ和Aggrecan的基因及蛋白含量表达明显降低,说明基质金属蛋白酶能很好反映膝关节软骨代谢变化,它对关节软骨基质具有降解作用,能加速软骨退变。
2针刀松解法能使关节软骨中MMP-3的基因及蛋白含量表达降低,基质中的Col-Ⅱ和Aggrecan的基因及蛋白含量表达升高,说明针刀松解法有一定的抑制细胞外基质胶原降解,保护促进和刺激软骨修复的作用,为针刀松解法治疗膝骨性关节炎提供了实验研究的依据,揭示了针刀松解法治疗膝关节骨性关节炎的科学内涵。
Osteoarthritis is a common degenerative joint disease, more common and frequently in the elderly, in three of the most common diseases by elderly patients (the most common cardiovascular disease, osteoporosis, osteoarthritis), the incidence rate of osteoarthritis in the first place in the world.Osteoarthritis occurs in the weight-bearing larger spine, knee, hip and other parts, and the main lesion is degeneration of articular cartilage and secondary bone hyperplasia, clinical manifestations of slowly progressive development of joint pain, swelling, stiffness, with limited mobility.Severely affected patients in daily life and work, and even can cause disability. With the population aging process accelerated, this problem becomes increasingly prominent. Therefore, the prevention and treatment of osteoarthritis has received widespread attention in the community, and the osteoarthritis research has important practical significance.Currently the etiology and pathogenesis of this disease is not clear.
Due to weight bearing of knee joint and activity of it is larger, so the knee osteoarthritis (KOA) is one of the most common bone arthritis, generally conservative treatment is more difficult and easy recurrence.For more than30years, acupotomy as a new, minimally invasive technique of traditional Chinese medicine,had better curative effect in the treatment of KOA, but basic research remains to be in-depth.This study through experiments to explore possible mechanisms of acupotomy for treating knee osteoarthritis, lay a foundation for further research, and provide theoretical basis for clinical treatment.
Objective
The experiment used acupotomy intervention and compared with electric acupuncture group, to observe acupotomy intervention on animal models of osteoarthritis articular cartilage tissue type Ⅱcollagen (Col-Ⅱ), aggrecan (Aggrecan), matrix metalloproteinases-3(MMP-3) gene and protein expression, to explore the possible mechanism of knee osteoarthritis acupotomy intervention from the perspective of molecular biology, to lay the foundation for the later research, and also provide a theoretical basis for clinical treatment of knee osteoarthritis.
Methods
Six-month-old healthy adult New Zealand rabbits(equal between genders) were divided into four groups randomly:normal control group, model group, acupotomy group,electroacupunctur(EA) group. The left hind limb immobilization in extension position braking method (Vidman method) for preparing animal models of knee osteoarthritis. Acupotomy group and electro-acupuncture group began to intervene after one week at the end of molding, acupotomy group intervention for one time per week, a total of three weeks of intervention, electric acupuncture group was treated one time on alternate days, treatment three times in one weeks, a total of three weeks of treatment. Before and after treatment knee joint behavior index were observed in rabbit; gross observed pathological manifestations of the articular cartilage and synovial membrane; pathology of rabbit cartilage slices were observed under optical microscope; Real-time PCR method to detect Col-ⅡmRNA, AggrecanmRNA, MMP-3mRNA expression level, and Western blot method to detect Col-Ⅱ,Aggrecan, MMP-3protein expression level.
Results
1General observation:Normal control group rabbits knee joints were not swelling, congestion, joint fluids transparent, articular cartilages pale blue, translucent, smooth. Synovial membranes was without edema, hypertrophy.Model group rabbits knee joints were significantly swelling or congestion. Joint fluid was pale yellow and slightly turbid.Articular cartilages were gray, dull, not smooth, and not opaque.There were defect.Ulcers and fissures could be seen clearly. Some subchondral bone was exposed, medial tibial condyle more obvious, synovial hyperemia, edema, hypertrophy.Acupotomy group rabbits the left knee joints mild swelling and had no significant congestion, cartilage pale color, bright color, mild defect, and the obvious ulcers was not seen.Joint fluid was transparent and slightly yellow.Compared with the model group,edema and hyperemia was significantly ease. EA group articular cartilages transparency reduced and were less shin. Some visible cracks could be seen.Synovial hyperplasia reduced,and there was not significant congestion.Indicating acupotomy, electro-acupuncture treatment can relieve cartilage damage,and joint edema.
2Optical microscope observation:Knee joint articular cartilages of normal control group were structural integrity.Articular cartilage surface was smooth.Thickness was normal. Cartilage cells arranged in neat rows, and layered clear, orderly. Cartilage matrix was homogencously,and tide line was clear and complete, no pannus formation. In model group, the cartilage surface was not smooth.The surface of cartilage cells were scarce.Cells arranged irregularly. Levels were disturbance.Sometimes visible peeling defect was as deep as the tide line. Chondrocytes nested.Tide line blurred or double tide line was seen, part of the cartilage pannus formation and vascular. Acupotomy group and electro-acupuncture group cartilage cells arranged in neat rows. Cartilage fracture was rare.But local cells arranging in disorder still can be seen. Repetitive tide line was occasionally seen.Cartilage surface was smooth,without pannus formation. The sections were scored according to Mankin method.Four groups overall score were significantly different (P<0.01). Acupotomy group comparing with model group had significant difference (P<0.01); and compareing with the control group had significant difference (P<0.05).There was not significant difference as compared with electroacupuncture group (P>0.05). EA group comparing with the control group had significant difference (P<0.01), and no significant difference as compared with the model group (P>0.05). Acupotomy method had cartilage protective effect.
3Behavior changes:the model group and acupotomy group, acupuncture group Lequesne MG score before treatment had no significant statistical difference (P>0.05).The score difference before and after treatment, and score after treatment had significant difference (P<0.05).Acupotomy therapy is effective in improving the rabbit KOA joint pain, swelling and gait, activity.
4Real-time PCR experimental results showed Col-ⅡmRNA content expression of acupotomy group increased more significantly than that of model group, and there were significant differences between the two (P<0.05). Compared with model group, Col-ⅡmRNA content expression of electro-acupuncture group significantly increased, and there were significant differences between them(P<0.05). Electroacupuncture group had significant difference with normal control group (P<0.05). Cartilage AggrecanmRNA content expression of model group was significantly decreased,compared with control group,and there were significant differences between the two compared (P<0.01).The expression of MMP-3mRNA in model group was significantly increased compared with normal control group, and there were significant differences between the two (P<0.05).Each group decreased as compared with the model group, but no significant difference compared with the normal control group (P>0.05).Acupotomy group decreased as compared with the model group, but no significant difference between the two (P>0.05). The results of Real-time PCR tests showed that acupotomy could effective up-regulated expression of Col-ⅡmRNA and AggrecanmRNA, down-regulated MMP-3mRNA expression, and the effect of acupotomy group was better than electroacupuncture group.
5Western blot experimental results showed Coll-Ⅱprotein expression in cartilage of model group significantly decreased than that of normal control group group, and there were significant differences between the two (P<0.01). Acupotomy group increased significantly compared with model group, the two had very significant difference (P<0.01). There was significant difference between acupotomy group and electroacupuncture group (P<0.05).The expression of aggrecan protein in acupotomy group was higher than that of model group, and there was significant differences between the two (P<0.05. Electroacupuncture group and model group had no significant difference (P>0.05). The expression of MMP-3protein in model group was significantly increased compared with normal control group, and there was significant differences between the two(P<0.01), Acupotomy group was significantly lower than the model group, and the two had significant differences(P<0.01).There was significant difference between acupotomy group and the electroacupuncture group (P<0.05). The results of Real-time PCR tests showed that acupotomy could effectively increase expression of Col-Ⅱprotein and Aggrecan protein,reduced expression of MMP-3protein, and the effect of acupotomy group was better than control group.
Conclusions
1Gene and protein expression of MMP-3in articular cartilage model group rabbits significantly increased, while the gene and protein content expression in the cartilage matrix of Coll-Ⅱand Aggrecan decreased obviously, sand howed that matrix metalloproteinases could well reflect the changes of knee cartilage metabolism, It had a role in the degradation of the extracellular matrix of articular cartilage, and could accelerate the degeneration of cartilage.
2Acupotomy can make the gene and protein content expression of MMP-3in articular cartilage decreased, and gene and protein content expression in the matrix of Col-Ⅱand Aggrecan elevated, showing that acupotomy was inhibition of extracellular matrix collagen degradation, protection to promote and stimulate cartilage repair function.Provided the basis for the experimental study of acupotomy treatment of osteoarthritis of the knee, and reveaed the scientific connotation of acupotomy in treating knee joint osteoarthritis.
引文
[1]李宁华.中老年人群骨关节炎的流行病学特征[J].中国临床康复,2005,9(38)：133-135.
[2]王章.中药治疗老年退行性膝关节骨性关节炎33例临床观察[J].江苏中药,2005,5(26)：26.
[3]杜以宽,马民,唐勇.中医对骨性关节炎病因病机的认识[J].浙江中医药大学学报,2009,33(2)：155.
[4]潘裕辉,周学平.浅谈中医对类风湿关节炎病因的认识[J].甘肃中医,2007,20(5).
[5]肖林榕.秦汉隋时期辨治骨痹(骨关节炎)理论的确立[J].福建中医学院学报,2004,14(2).
[6]黄俊卿.论《诸病源候论》的痹病理论体系[J].河南中医药学刊,1997,(6)：2.
[7]张玉莲,桑希生.浅析历节病与骨痹[J].中医药学报,2011,39(1)：5-7.
[8]王兴华整理,张氏医通[M].北京：人民卫生出版,2006：341(1)：51-52.
[9]黄斌,先静.《外台秘要》外来药物的考察[J].中医药学报,1990,1(1)：51-52.
[10]王亚芬.元《御药院方》中有关香药的临床应用[J].中国中药杂志,1995,20(3)：187-189.
[11]王衮.博济方[M].北京：商务印书馆,1959：2.
[12]王亚芬.《杨氏家藏方》几种病证中虫类药的应用[J].中医杂志,1997,38(4)：243-244.
[13]章健,李洪涛.《太平惠民和剂局方》方剂特点分析[J].中国医药学报,2002,17(4)：212-215.
[14]曲艺,肖林榕.唐宋元医家治疗骨痹的用药特点[J].福建中医学院学报,2008,18(2)：51-52.
[15]张燕霞,张成博.骨痹古代中医文献研究[J].山东中医药大学学报,2012,36(3).
[16]岑祖怡,周剑鹏.独活寄生汤治疗膝关节骨性关节炎50例疗效观察[J].新中医,2010,42(8)：50-51.
[17]孙书龙,汤晓晨,姜宏.“芪藤汤”治疗膝骨关节炎例临床观察[J].江苏中医药,2012,44(11)：45.
[18]王正强,周驱,宁安等.除痹汤治疗膝骨关节炎临床报告[J].中国中医骨伤科杂志,2012,20(1)：47.
[19]乌日图,付准,郑玲玲.刺老苞胶囊治疗膝关节骨性关节炎疗效观察[J].中药药理与临床,2012,28(5)：224-225.
[20]钱海平,罗旭耀,施水潮.健步关节胶囊治疗膝关节骨性关节炎疗效观察[J].海军医学杂志,2012,33(6)：398-399.
[21]仲跻申,张栋梁.消瘀通膝汤熏蒸治疗膝骨性关节炎血瘀证疗效观察[J].吉林中医药,2012,32(11)：1124.
[22]王超英,李迎新.白金散穴位敷贴结合TDP神灯治疗膝骨关节炎疗效观察[J].广州医药,2011,42(1)：46-47.
[23]包力,徐新玉,卓鹰.温痹散场效应热敷疗法治疗膝骨关节炎47例[J].中国中医骨伤科杂志,2012,20(1)：49.
[24]童国伟.海桐皮汤熏洗治疗重度膝骨关节炎疗效观察[J].上海中医药杂志,2012,46(6)：60.
[25]刘鑫,卢敏,谢心军.独活寄生汤合活血安痛酒离子导入治疗膝骨性关节炎30例[J].湖南中医杂志,2012,28(3)：71.
[26]林潮炫.灯盏花素对促进骨性关节炎治愈的临床疗效观察[J].吉林医学,2013,34(6)：1073.
[27]付宏伟,尚春香,陈晓华等.丹红注射液用于治疗膝关节骨性关节炎的疗效观察[J].中国医药指南,2012,10(21)：259.
[28]宋锦华.中药内外结合治疗膝关节骨性关节炎伴关节积液83例[J].中国医药指南,2011,9(2)：122.
[29]杨延国.中药内服外敷与塞来昔布治疗膝骨关节炎临床疗效对比观察[J].时珍国医国药,2012,23(10)：2640.
[30]殷光涛,胡英,王锦.针灸疗法治疗膝关节骨性关节炎100例[J].中国实用医学杂志,2010,20(10)：26-27.
[31]赵维杰.火针配合中药治疗膝骨关节炎[J].中国民族民间医药：167.
[32]顾力军,裘兴栋,赵勇等.铍针与扶他林乳胶剂治疗膝骨关节炎疼痛的病例对照研究[J].中国骨伤,2012,25(12)：1036.
[33]吴明清,罗启年.刃针治疗膝骨性关节炎的疗效及对X片生物力学影响的临床研究[J].实用疼痛学杂志,2011,7(4)：261-263.
[34]田有粮,李月,李茜等.长圆针治疗对膝关节骨性关节炎的作用[J].中国康复,2010,25(6)：444-445.
[35]马楚南,冯小敏,冯爱竹.小针刀治疗膝关节骨性关节炎的疗效分析[J].青海医药杂志,2008,38(7)：14.
[36]赵长伟,梁运海,闻辉等.手法治疗膝关节骨性关节炎疗效观察[J].长春中医药大学学报,2012,28(6)：1086.
[37]亓夫国,卞雪霞察.抗骨质增生膏贴敷配合推拿治膝关节骨性关节炎60例[J].中国民间疗法,2013,21(1)：28.
[38]何铭涛,李颖,邵敏.内服中药配合推拿治疗膝骨性关节炎的观察[J].中国中医骨伤科杂志,2007,15(8)：25.
[39]杨庆宇.独活寄生汤配合电针TDP治疗膝关节骨性关节炎180例[J].中国社区医师,2013,15(4)：224.
[40]胡德,吴健武,黄光隆.膝痹通联合关节内注射玻璃酸钠治疗膝关节骨性关节炎[J].按摩与导引,2006,22(3)：18.
[41]王远军,罗吉祥.小针刀联合玻璃酸钠曲安奈德注射治疗膝骨性关节炎885例[J].贵州医药,2012,36(10)：945-946.
[42]多果.藏医用药酒治疗关节炎的体会[J].中国民族民间医药杂志,1999：24-25.
[1]游苏宁.膝骨关节炎的流行病学调查[J].中华内科杂志,1995,34(2).
[2]曾庆徐,黄少弼,肖征宇.症状性骨关节炎的临床和流行病学探讨[J].中华内科杂志,1995,34(2)：88-90.
[3]王伟,王坤正,党小谦等.中老年人人群骨关节炎的流行病学研究[J].中国老年学杂志,2007,27：566.
[4]向珍蛹,茅建春,徐先国等.膝骨关节炎中医证型分布的流行病学研究[J].上海中医药杂志,2012,46(12)：5.
[5]林华,陈新,张咏梅等.绝经后妇女骨质疏松与骨关节炎的相关性研究[J].中国骨质疏松杂志,2006,12(5)：488-489.
[6]Rego-Perez I, Fernandez-Moreno M, Fernandez-Lopez C, et al. Mitochondrial DNA haplo groups:role in the prevalence and severity of knee osteoarthritis[J]. Arthritis Rheum,2008,58(8):2387-2396.
[7]Page G P, George V, Go R C, et al."Are we there yet?":Deciding when one has demonstrated specific genetic causation in complex diseases and quantitative traits[J].Am J Hum Genet,2003,73(4):711-719.
[8]Valdes A M, Loughlin J, Chapman K, et al. Sex and ethnic differences in the associationofASPN,CALM1,COL2A1,COMP,and FRZB with genetic susceptibility to osteoarthritis of the knee[J]. Arthritis Rheum,2007,56(1):137-146.
[9]李光第,白松,宋恩.膝关节骨性关节炎病因及诊治的[J]哲学思维医学与哲学2012,33：54.
[10]刘献祥,林燕萍.中西医结合治疗骨性关节炎.第一版[M].北京：人民卫生出版社,2009：38-39.
[11]Mcalindon TE,Jacques P,Zhang-Y,et al.Do antioxidant micronutrients protect against the development and progression of knee osteoarthritis[J].Arthritis Rheum,1996,39:648-656.
[12]伦学刚,谈志龙,白人骁.骨质疏松与骨性关节炎关系的研究进展[J].中国骨伤,2007,20(12)：876-877.
[13]Bergink AP,Uitterlinden AQVan Leeuwen JP, et al. Bonemineral density and vertebral fracture history are associated with incident and progressive radiographic knee osteoarthritis in elderly men and women:the rotter dam study. Bone,2005,37:446-456.
[14]ElMiedany YM,Mehanna AN, El Baddini MA. Altered bone mineral metabolism in patients with osteoarthritis. Joint Bone Spine,2000,67.
[15]黄火高,郑毅,尹义存.骨性关节炎的定量超声骨密度研究[J].天津医药,2004,32 (12)：737-739.
[16]Castaneda S,Largo R, Calvo E,et al.Bone mineral measurement of subchondral and trabecular bone in healthy and osteoporotic rabbits. Skeletal Radiol,2006,35 (1) 34-41.
[17]Apparailly F, Verwaerde C, Jacquet C, et al. Adenovirus-mediated transfer of viral IL-10 gene inhibits murine collagen-induced arthritis[J].Immuneol,1998,160(11):5213-5220.
[18]Haynes MK,Hume EL,Smith JB.Phenotypic characterization of inflammatory cells from ostoarthritic synovium and syovial fluids[J].Clin Immunol,2002,105(3): 315-325.
[19]李宁华.中老年人群骨关节炎的流行病学特征[J].中国临床康复,2005,38(10)：133-135.
[20]Waine H Nevinny D Rosenthal J Joffe IB:Association of osteoarthritis and diabetes mellitus.Tufts Folia Med 1961,7:13.
[21]韩凯,熊昌源.“三联疗法”对兔膝关节骨关节炎氧自由基代谢的影响[J].湖北中医杂志,2006,28(3)：10-11.
[22]赵洪普,郭玉海.骨关节炎与细胞因子的研究进展[J].中华综合医学杂志,2003,5(8)：85.
[23]Kaurg, HalliwellB.Evidence for Nitrie Oxide mediated oxidat-ived maageine hronie inflammation.Nitro tyrosine in serum and-synovia fluid from rheumatoid Patients.FEBSL et,1994,35O:9-12.
[24]M Rossignol, A Leclerc, FAAllaert, et al.Primary osteoarthritis of hip knee, and hand in relation to occupational exposure[J].Occup Environ Med,2005,62:772-777'.
[25]查建林.膝骨关节炎的病因与生物力学的关系[J].医药论坛,2009,(1)：35-36.
[26]Oliveria SA, Caffrey M, Linden SM, et al.The analysis of epidemiology about human arthritis[J].Epidemiology,2004,15:326-331.
[27]王序全,李起鸿,机械应力异常与关节软骨退变[J].国外医学生物医学工程分册,1999,22(2)：100-192.
[28]吕存贤,王维佳.膝骨关节炎病因病理研究及中医治疗概况[J].浙江中医学院学报,2005,29(1)：82.
[29]李忆农.细胞因子与关节炎[J].中华风湿病学杂志,2000,24(1)：56-58.
[30]Neidel J,sihulze M,SoveL,et at.Practical significance of cytokine determi-nation in joint Fluid patients with arthroses or rheu-matoid arthritis[J].Z Orthop Ihre Grenzgeb, 1996,(134):381.
[31]顾迁,戴戎,裘世静等.异常高应力导致关节软骨退变机理的形态学研究[J].中华外科杂志,1995,33(10)：597-600.
[32,33]王序全,李起鸿.机械应力异常与关节软骨退变[J].国外医学生物医学工程分册1999,22(2)：100-192.
[34]高翔.益气化瘀利水方干预兔膝骨关节炎软骨中胰岛素样生长因子1的变化[J].中国临床康复,2006,10(39)：95-97.
[35]Lauder SN,Carty SM,Carpenter CE, et al. Interleukinlbeta induced activation of nuclear factor kappab can be inhibited by no el pharmacological agents in osteoarthritis. Rheumatology(Oxford),2007,46(5):752-758.
[36]Sakimura K, Matsumoto T, Miyamoto C, et al.Effects of insulin like growth fact or I on transforming growth factor betal induced chondrogenesis of synovium derived mesenchymal stem cells cultrued in a polyglycolic acid scaffold. Celles T issues Organs,2006,183 (2):55-61.
[37]Blaney Davidson EN, van der Kraan PM,van den Berg WB. TGF beta and osteoarthritis. Osteoarthritis Cartilage,2007,15(6):597-604.
[38]Blaney Davidson EN, Scharstuhl A, Vitters EL, eal. Reduced transforming growth factor-beta signaling in cartilage of old mice:role in impaired repair capacity. Arthritis Res T her,2005,7 (6):R1338-R1347.
[39]van Beuningen HM, van der Kraan PM, Arntz OJ,et al. Transforming growth factor-beta 1 stimulate articular chondrocyte proteoglycan synthesisand induces o steophyte formation in the murine knee joint Lab Invest,1994,71(2):279-290.
[40]Malemud CJ, Killeen W, H ering T M, et a 1. Enhanced sulfated proteogly can core protein synthesis by incubation of rabbit chondrocytes with recombinant transforming growth factor bet a l. J Cell Physiol,1991,149(1):152-159.
[41]Iwanaga H, M atsumoto T, Enomoto H, et al. Enhanced expression of insulin like growth factor binding proteins in human osteoarthritic cartilage detected by immunohistochemistry and in situ hybridization.Osteoarthritis Cartilage,2005, 13(5):439-448.
[42]Barta E, M aroudas A. A theoretical study of the distribution of insulin like g rowth factor in human articular cartilage. J T heor Biol,2006,241(3):628-638.
[43]张德辉,薛刚,黄昌林.应用玻璃酸钠对关节镜术后膝骨关节炎功能恢复的影响[J].中国临床康复,2002,12(6)：1730-1731.
[44]乔聚义,李秋莉,席彬等.低场强MRI对椎体终板骨软骨炎的诊断价值[J].中华实用诊断与治疗杂志.2010,24(11)：1103-1104.
[45]Jasin H E. Autoantibody specificities of immune complexes sequestered in articular cartilage of patients with rheumatoid arthritis and osteoarthritis.Arthritis Rheum,1985, 18:241.
[46]Herman JH, Appel AM.Immunologic modulation of cartilage metabolism.In:Pelletier JP.Edited Proteases:The or involvement in OA. J Rheumatol,1987,14,(Supp 1)1483.
[47]王新文,张民,卫小春.膝骨关节炎发病的免疫机制研究进展[J].中国现代医生,2010,48(18)：16-18.
[48]Burnett BP, Levy R, Cole BJ. Metabolic mechanisms in the pathogenesis of osteoarthritis.[J].Knee Surg,2006;19(3):191-197.
[49]Janet M, Anderson—MacKenzie, Helen—L et al.Fundamental subchondral bone changes in spontaneous knee osteoarthritis[J].The international Journal of Biochemistry& Cell biology,2004,(10):1-13.
[50]李强,张柳.骨性关节炎中软骨下骨与骨退变的关系研究进展[J].中国修复重建外科杂志,2009,23(2)：245-247.
[1]徐叔云,卞如濂,陈修.药理实验方法学.第3版.[M].北京：人民卫生出版社,2002：926.
[2]郭建刚等.骨性关节炎软骨的生化及代谢变化[J].中华骨伤杂志,1998,11(5)：73-74.
[3]李茂强,史晨辉,王永明.骨关节炎的实验研究现况与进展[J].农垦医学2004,26(1)：29.
[4]Aigner T, Sachse A, Gebhard PM, et al.Osteoarthritis:Patho-biology-targets and ways for therapeutic intervention[J]. Adv Drug Delivery Rev,2006,58 (2):128-149.
[5]Gold ring MB. Update on the biology of the chondrocyte and new approaches to treating cartilage diseases[J].Best Pract Res Clin Rheumatol,2006,20(5):1003-1025.
[6]Mar oudasA. Balance betweens welling pressure and collagen tension in and normal degenerate cartilage [J]. Nature,1976,260(5554):808-809.
[7]Fosang AJ, Rogers on FM, East CJ, et al. ADAMTS2 5:The story so far[J]. Eur Cell Mater,2008,15(1):11-26.
[8]PrattaMA, YaoWQ, Decicco C, et al. Aggrecan protects cartilage collagen from proteolytic cleavage[J]. J Biol Chem,2003,278(46):45539-45545.
[9]Nagase H, Woessner JF Jr. Matrix metallopromteinases [J]. J Boil Chem 1999,274(3):21491-21494.
[10]Yu LP Jr. Smith GN Jr.HastykA.et al.Doxycycllne inhibits type II Collagen olytic activity of extracts from human osteoarthritic Cartilage and of gelatinase.J Rheumatol,1991,18(10):1450-1452..
[11]Murphy G, Knauper V, Atkinson, et al. Matrix metalloproteinases in arthritic disease.Arthriistes,2002,4(Suppl 3):39-49.
[12]Lijnen HR.Elements of the fibrinolytic system[J].Ann NY Acad Sci,2001,936:226-36.
[13]徐海涛,陈游,陈立科,不同干预因素对膝骨关节炎患者滑液中MMP-3和TIMP-1的影响[J].中南大学学报(医学版),2008,33(1)47.
[14]Taylor DJ, Cheung NT, Dawes PT. Increased serum pro-MMP-3 in inflammatory arthritides:a potential indicator of synovial inflammatory monokie activity[J].Annn Rheum Dis,1994,53(11):768-772.
[15]Ichikawa Y, Yamada C,Hori ki T,et al.Serum matrix metalloproteinse-3 and fibrin degradation product levels correlate with clinical disease activity in rheumatoid arthritis[J].Clin Exp Rheumatol,1998,16 (5):533-54.
[16]王晓霞,李小峰,王来远等.类风湿关节炎血清基质金酶-3水平及其影响因素的探讨[J].中华风湿病学杂志,(1)：16-19.
[17]魏平,付爽,王俊祥.基质金属蛋白酶3在类风湿关节炎达及其意义[J].临床荟萃,2006,21(19)：1389-1390.
[18]朱宝玉,陈琼,王万春.基质金属蛋白酶-3(MMP-3)在创伤后关节软骨退变中的作用[J].实用骨科杂志.2006,12(1)：35.
[19]陈希,涂长青,刘丽娟.补肾活血方对膝骨关节炎蛋白聚糖和Ⅱ型胶原表达的影响[J].新中医,2008,40(8)：47.
[20]梁祖建,韩清民,张还添.补肾活血方对骨关节炎关节软骨保护的效应与机制[J].江苏中医药,2009,41(3)：34.
[21]江艳,李荣亨.复元胶囊对膝骨关节炎患者血清MMP-3和TIMP-1表达变化的影响[J].西南国防医药,2010,20(9)：934.
[22]李超等.蛋白聚糖酶和骨关节炎.[J].北京大学学报(医学版),2009,41：611-613.
[23]Sh arif M, Kir wan J, Cham iN, et al A 5-yr longitudinal study of type Ⅱ A collagen synthesis and total type Ⅱ collagen degradation in patients with knee osteoarthritis-association with disease progression [J]. Rheumatology,2007,46:938-943.
[24]Sh inj iU, Yos h i h it o N, Yas um asa S, et a.l Fluid Levels of Chon-droit in Sulfate in Patients with Osteoarthritis of the Knee Joint[J].JN ppon Med Sch,2001,68:165-170.
[1]刘向前.中西医结合治疗膝关节骨关节炎(第1版)[M].人民军医出版社,2011：23.
[2-3]刘献祥,林燕萍.中西医结合治疗膝关节骨关节炎(第1版)[M].人民卫生出版社,2009：3.
[4]张蓉,李峰,王常海.经筋理论在膝关节骨性关节炎发病机制及治疗中的作用[J].中国康复医学杂志,2007,22(7)：644.
[5]任平霞.综合治疗早中期膝关节骨性关节炎169例[J].光明中医,2011,26(8)：1631-1632.
[6]Videman T. Experimental osteoarthritis in the rabbit:comparison of different periods of repeated immobilization [J].Acta Orthop Scand,1982,53(3):339-347.
[7]Salo PT, Hogervorst T, Seerattan RA. Selective joint denervation promotes knee osteoarthritis in the aging rat [J].J orthop Res,2002,20(6):1256-1264.
[8]Yamamoto K Yamamoto K, Shishido T, Masaoka T, et al. Morphologicalstudies on the ageing and osteoarthritis of the articular cartilage in C57 black mice [J]. Orthop Surg (kong Kong),2005,13(1):8-18.
[9]刘伯龄,刘献祥.骨性关节炎动物模型的建立概况[J].福建中医药,2003,34(4)：49.
[10]杨峰,史索道.用木瓜蛋白酶建立兔颚须关节骨关节炎模型[J].华西口腔医学杂志,2002,12(5)：330-333.
[11]Kikuchi T, Sakuta T, Yamaguchi T. Intra-articular injection of collagenase induces experimental osteoarthritis in mature rabbits[J].Osteoarthritis Cartilage, 1998,6(3):177-186.
[12]T sai CL,L iu TK. Estradiol-induced knee osteoarthrosis in ovariectomized rabbits[J].Clin Orthop,1993,(291):295-302.
[13]Sakano Y, Terada N,U eda H, et al. Histo logical study of articular cartilage in experimental rat knee arthritis induced by intracapsular injection of cationic poly ethleneimine[J].Med Electron Micro sc,2000,33(4):246-257.
[14]膝关节骨关节炎动物模型的构建[J].中国中医骨伤科杂志,2012,20(12)：71.
[15]M cDevitt CA, Gilbertson EMM. M uir H. An experimental model of osteoarthritis. Early morphological and biochemical changes[J].J bone Joint Surg,1977,59.(B):24-30.
[16]顾廷,戴克戎,裘世静等.应力降低导致关节软骨退变机理形态学研究[J].中华骨科杂志,1995,15(9)：631-633.
[18]M arijnissen AC, Van Roermund PM, Te Koppele JM, et al. The canine'groove'model, compared with the ACL T model of osteoarthritis[J].Osteoarthritis Cartilage,2002,10(2):145-155.
[19]白希壮,任继尧.选择性臀肌切断诱发骨关节炎实验模型[J].中华骨科杂志,1994,14(2)：118-120.
[20]刘蜀彬,孔祥星,史迎昌等.股骨干骺端髓内血运阻断诱发兔膝关节骨性关节炎模型[J].中国矫形外科杂志,2003,11(24)：1706-1707.
[21]胡波,胡志俊,张有为等.骨关节炎动物模型：人工诱导与动物自发[J].中国组织工程研究与临床康复,2010,14(33)：6202-6205.
[22]刘媛,赵驻军,王红芹等.寒冷刺激鸡膝关节软骨显微结构变化观察[J].河北农业大学学报,2008,31(2)：93-95.
[23]陆志华,姚伟武,胡曙光等应用磁共振生理学成像定量测定兔膝关节软骨退变的研究[J].医学影像学杂志,2009,19(6)：739-744.
[24]王学谦,娄思权,侯筱魁等译.骨关节炎诊断与治疗(第四版)[M].天津科技翻译出版公司,2008,3：59-77.
[25]邱宜兴兔关节制动引起关节软骨退变作用的实验研究[J].中华骨科杂志.1987；25(3)：175.
[26]吴海山,周维江,张中南.老年性退变膝[J].中华老年医学杂志,1994,13(6)：330.
[27]吴毅,范振华,屠丹云等.运动促进运动后兔膝关节组织修复的应用[J].中国康复医学杂志,1993,8(1)：17-20.
[28]祖启明,王庆良,郭正成.关节持续被动运动的基础研究和临床应用[J].国外医学物理医学与康复学分册,1995,15(1)：155-158.
[29]邱贵兴,王桂生.兔膝关节制动引起关节软骨退变的实验研究[J].中华外科杂志,1987,25(30)：175-177.
[1]Lequesne MG, Samsons M. Indices of severity in osteoarthritis of weight bearing joints [J].JRheumatol Supp,1991,27:16-18.
[2]Lequesne MG, Mery C, Samson M, et al. Comparison between the WOMAC and the Lequesne indices in patients with knee and hip osteoarthritis [J].Osteoarthritis Cartilage, 1998,6(6):441-442.
[3]Lequesne MG, Maheu E. Methodology of clinical trials in hand osteoarthritis: conventional and proposed tools [J]. Osteoarthritis Cartilage,2000,8(Suppl A):64-69.
[4]Lequesne M, Maheu E, Cadet C, et al. Structural effect of avocado/soybean unsaponifiables on joint space loss in osteoarthritis of the hip [J]. Arthritis Rheum,2002, 47(1):50-58.
[5]Lequesne MG, Maheu E. Clinical and radiological evaluation of hip, knee and hand osteoarthritis [J]. Aging Clin Exp Res,2003,15(5):380-390.
[6]龚权,程传国,程龙.药物电针刀疗法对兔膝骨性关节炎软骨细胞凋亡的影响[J].中国中西医结合杂志基础理论研究特集,2004,24：195-196.
[7]曾贵刚,张秀芬,权伍成。针刀松解法对膝骨性关节炎应力刺激及临床症状的影响[J].中华中医药学刊,2009,27(1)：66-68.
[8]McCarry DJ:Arthritis and Allied Condition1Oth ed, Lea and Febiger, PhiladelPhia,1985.
[9]Doyle DV:J pathol,1982,136(3):199.
[10]BurgerC:Acta Orthop Scand,1983,54(1):132.
[11]Egan MC:Arhritis Rheum,1982,25(2):204.
[12]InoueH:Int Orthop,1981,5(1):47.
[13]Finsterbush A:Clin Orthop,1977,111:290.
[14]Palmoski M:Arthritis Rheum.1981,24(10):1329.
[15]Fabry G:Clin Orthop,1980,153:253.
[16]Radin EL:J Biomech,1973,6(1):51.
[17]Sapolsky AI:Arthritis Rheum.1982,25(8):981.
[18]Chadilly FN:Fine Structure of Synovial Joints Butterworth,London,1983.
[19]朱汉章主编.针刀医学.第1版.[M].北京：中国中医药出版社,2004：128.
[1]徐叔云,卞如濂,陈修.药理实验方法学[M].3版.北京：人民卫生出版社,2002：926.
[2]Aigner T, Sachse A, Gebhard PM, et al. Osteoarthritis:Patho-biology-targets and ways for therapeutic intervention[J]. Adv Drug Delivery Rev,2006,58 (2):128-149.
[3]Gold ringMB. Update on the biology of the chondrocyte and new approaches to treating cartilage diseases[J]. Best Pract Res Clin Rheumatol,2006,20 (5):1003-1025.
[4]Mar oudasA. Balance bet ween s welling p ressure and collagen tension in normal and degenerate cartilage [J]. Nature,1976,260(5554):808-809.
[5]Fosang AJ, Rogers on FM, East CJ, et al. ADAMTS2 5:The story so far[J]. Eur Cell Mater,2008,15(1):11-26.
[6]PrattaMA, YaoWQ, Decicco C, et al. Aggrecan p rotects cartilagecollagen from pr oteolytic cleavage [J]. J Biol Chem,2003,278(46):45539-45545.
[7]Nagase H, Woessner JF Jr. Matrix metallopromteinases [J]. J Boil Chem,1999,274(3): 21491-21494.
[8]Yu LP Jr. SmIth GN Jr.HastykA.et al.Doxycycllne inhibits type Ⅱ Collagenolytic activity of extracts fromhuman osteoarthritic Cartilage and of gelatinase.J Rheumatol,1991, 18(10):1450-1452.。
[9]Murphy G, Knauper V, Atkinson, et al. Matrix metalloproteinases in arthritic disease. Arthriistes,2002,4(Su ppl 3):39-49.
[10]LijnenHR.Elements of the fibrinolytic system[J].Ann NY Acad Sci,2001,936:226-36.
[11]徐海涛,陈游,陈立科.不同干预因素对膝骨关节炎患者滑液中MMP-3和TIMP-1的影响[J].中南大学学报(医学版),2008,33(1)：47.
[12]庞坚,曹月龙,詹红生.骨关节炎的“筋”“骨”之辨[J].上海中医药大学学报,2012,26(1).
[2]王章.中药治疗老年退行性膝关节骨性关节炎33例临床观察[J].江苏中药,2005,5(26)：26.
[3]杜以宽,马民,唐勇.中医对骨性关节炎病因病机的认识[J].浙江中医药大学学报,2009,33(2)：155.
[4]潘裕辉,周学平.浅谈中医对类风湿关节炎病因的认识[J].甘肃中医,2007,20(5).
[5]肖林榕.秦汉隋时期辨治骨痹(骨关节炎)理论的确立[J].福建中医学院学报,2004,14(2).
[6]黄俊卿.论《诸病源候论》的痹病理论体系[J].河南中医药学刊,1997,(6)：2.
[7]张玉莲,桑希生.浅析历节病与骨痹[J].中医药学报,2011,39(1)：5-7.
[8]王兴华整理,张氏医通[M].北京：人民卫生出版,2006：341(1)：51-52.
[9]黄斌,先静.《外台秘要》外来药物的考察[J].中医药学报,1990,1(1)：51-52.
[10]王亚芬.元《御药院方》中有关香药的临床应用[J].中国中药杂志,1995,20(3)：187-189.
[11]王衮.博济方[M].北京：商务印书馆,1959：2.
[12]王亚芬.《杨氏家藏方》几种病证中虫类药的应用[J].中医杂志,1997,38(4)：243-244.
[13]章健,李洪涛.《太平惠民和剂局方》方剂特点分析[J].中国医药学报,2002,17(4)：212-215.
[14]曲艺,肖林榕.唐宋元医家治疗骨痹的用药特点[J].福建中医学院学报,2008,18(2)：51-52.
[15]张燕霞,张成博.骨痹古代中医文献研究[J].山东中医药大学学报,2012,36(3).
[16]岑祖怡,周剑鹏.独活寄生汤治疗膝关节骨性关节炎50例疗效观察[J].新中医,2010,42(8)：50-51.
[17]孙书龙,汤晓晨,姜宏.“芪藤汤”治疗膝骨关节炎例临床观察[J].江苏中医药,2012,44(11)：45.
[18]王正强,周驱,宁安等.除痹汤治疗膝骨关节炎临床报告[J].中国中医骨伤科杂志,2012,20(1)：47.
[19]乌日图,付准,郑玲玲.刺老苞胶囊治疗膝关节骨性关节炎疗效观察[J].中药药理与临床,2012,28(5)：224-225.
[20]钱海平,罗旭耀,施水潮.健步关节胶囊治疗膝关节骨性关节炎疗效观察[J].海军医学杂志,2012,33(6)：398-399.
[21]仲跻申,张栋梁.消瘀通膝汤熏蒸治疗膝骨性关节炎血瘀证疗效观察[J].吉林中医药,2012,32(11)：1124.
[22]王超英,李迎新.白金散穴位敷贴结合TDP神灯治疗膝骨关节炎疗效观察[J].广州医药,2011,42(1)：46-47.
[23]包力,徐新玉,卓鹰.温痹散场效应热敷疗法治疗膝骨关节炎47例[J].中国中医骨伤科杂志,2012,20(1)：49.
[24]童国伟.海桐皮汤熏洗治疗重度膝骨关节炎疗效观察[J].上海中医药杂志,2012,46(6)：60.
[25]刘鑫,卢敏,谢心军.独活寄生汤合活血安痛酒离子导入治疗膝骨性关节炎30例[J].湖南中医杂志,2012,28(3)：71.
[26]林潮炫.灯盏花素对促进骨性关节炎治愈的临床疗效观察[J].吉林医学,2013,34(6)：1073.
[27]付宏伟,尚春香,陈晓华等.丹红注射液用于治疗膝关节骨性关节炎的疗效观察[J].中国医药指南,2012,10(21)：259.
[28]宋锦华.中药内外结合治疗膝关节骨性关节炎伴关节积液83例[J].中国医药指南,2011,9(2)：122.
[29]杨延国.中药内服外敷与塞来昔布治疗膝骨关节炎临床疗效对比观察[J].时珍国医国药,2012,23(10)：2640.
[30]殷光涛,胡英,王锦.针灸疗法治疗膝关节骨性关节炎100例[J].中国实用医学杂志,2010,20(10)：26-27.
[31]赵维杰.火针配合中药治疗膝骨关节炎[J].中国民族民间医药：167.
[32]顾力军,裘兴栋,赵勇等.铍针与扶他林乳胶剂治疗膝骨关节炎疼痛的病例对照研究[J].中国骨伤,2012,25(12)：1036.
[33]吴明清,罗启年.刃针治疗膝骨性关节炎的疗效及对X片生物力学影响的临床研究[J].实用疼痛学杂志,2011,7(4)：261-263.
[34]田有粮,李月,李茜等.长圆针治疗对膝关节骨性关节炎的作用[J].中国康复,2010,25(6)：444-445.
[35]马楚南,冯小敏,冯爱竹.小针刀治疗膝关节骨性关节炎的疗效分析[J].青海医药杂志,2008,38(7)：14.
[36]赵长伟,梁运海,闻辉等.手法治疗膝关节骨性关节炎疗效观察[J].长春中医药大学学报,2012,28(6)：1086.
[37]亓夫国,卞雪霞察.抗骨质增生膏贴敷配合推拿治膝关节骨性关节炎60例[J].中国民间疗法,2013,21(1)：28.
[38]何铭涛,李颖,邵敏.内服中药配合推拿治疗膝骨性关节炎的观察[J].中国中医骨伤科杂志,2007,15(8)：25.
[39]杨庆宇.独活寄生汤配合电针TDP治疗膝关节骨性关节炎180例[J].中国社区医师,2013,15(4)：224.
[40]胡德,吴健武,黄光隆.膝痹通联合关节内注射玻璃酸钠治疗膝关节骨性关节炎[J].按摩与导引,2006,22(3)：18.
[41]王远军,罗吉祥.小针刀联合玻璃酸钠曲安奈德注射治疗膝骨性关节炎885例[J].贵州医药,2012,36(10)：945-946.
[42]多果.藏医用药酒治疗关节炎的体会[J].中国民族民间医药杂志,1999：24-25.
[1]游苏宁.膝骨关节炎的流行病学调查[J].中华内科杂志,1995,34(2).
[2]曾庆徐,黄少弼,肖征宇.症状性骨关节炎的临床和流行病学探讨[J].中华内科杂志,1995,34(2)：88-90.
[3]王伟,王坤正,党小谦等.中老年人人群骨关节炎的流行病学研究[J].中国老年学杂志,2007,27：566.
[4]向珍蛹,茅建春,徐先国等.膝骨关节炎中医证型分布的流行病学研究[J].上海中医药杂志,2012,46(12)：5.
[5]林华,陈新,张咏梅等.绝经后妇女骨质疏松与骨关节炎的相关性研究[J].中国骨质疏松杂志,2006,12(5)：488-489.
[6]Rego-Perez I, Fernandez-Moreno M, Fernandez-Lopez C, et al. Mitochondrial DNA haplo groups:role in the prevalence and severity of knee osteoarthritis[J]. Arthritis Rheum,2008,58(8):2387-2396.
[7]Page G P, George V, Go R C, et al."Are we there yet?":Deciding when one has demonstrated specific genetic causation in complex diseases and quantitative traits[J].Am J Hum Genet,2003,73(4):711-719.
[8]Valdes A M, Loughlin J, Chapman K, et al. Sex and ethnic differences in the associationofASPN,CALM1,COL2A1,COMP,and FRZB with genetic susceptibility to osteoarthritis of the knee[J]. Arthritis Rheum,2007,56(1):137-146.
[9]李光第,白松,宋恩.膝关节骨性关节炎病因及诊治的[J]哲学思维医学与哲学2012,33：54.
[10]刘献祥,林燕萍.中西医结合治疗骨性关节炎.第一版[M].北京：人民卫生出版社,2009：38-39.
[11]Mcalindon TE,Jacques P,Zhang-Y,et al.Do antioxidant micronutrients protect against the development and progression of knee osteoarthritis[J].Arthritis Rheum,1996,39:648-656.
[12]伦学刚,谈志龙,白人骁.骨质疏松与骨性关节炎关系的研究进展[J].中国骨伤,2007,20(12)：876-877.
[13]Bergink AP,Uitterlinden AQVan Leeuwen JP, et al. Bonemineral density and vertebral fracture history are associated with incident and progressive radiographic knee osteoarthritis in elderly men and women:the rotter dam study. Bone,2005,37:446-456.
[14]ElMiedany YM,Mehanna AN, El Baddini MA. Altered bone mineral metabolism in patients with osteoarthritis. Joint Bone Spine,2000,67.
[15]黄火高,郑毅,尹义存.骨性关节炎的定量超声骨密度研究[J].天津医药,2004,32 (12)：737-739.
[16]Castaneda S,Largo R, Calvo E,et al.Bone mineral measurement of subchondral and trabecular bone in healthy and osteoporotic rabbits. Skeletal Radiol,2006,35 (1) 34-41.
[17]Apparailly F, Verwaerde C, Jacquet C, et al. Adenovirus-mediated transfer of viral IL-10 gene inhibits murine collagen-induced arthritis[J].Immuneol,1998,160(11):5213-5220.
[18]Haynes MK,Hume EL,Smith JB.Phenotypic characterization of inflammatory cells from ostoarthritic synovium and syovial fluids[J].Clin Immunol,2002,105(3): 315-325.
[19]李宁华.中老年人群骨关节炎的流行病学特征[J].中国临床康复,2005,38(10)：133-135.
[20]Waine H Nevinny D Rosenthal J Joffe IB:Association of osteoarthritis and diabetes mellitus.Tufts Folia Med 1961,7:13.
[21]韩凯,熊昌源.“三联疗法”对兔膝关节骨关节炎氧自由基代谢的影响[J].湖北中医杂志,2006,28(3)：10-11.
[22]赵洪普,郭玉海.骨关节炎与细胞因子的研究进展[J].中华综合医学杂志,2003,5(8)：85.
[23]Kaurg, HalliwellB.Evidence for Nitrie Oxide mediated oxidat-ived maageine hronie inflammation.Nitro tyrosine in serum and-synovia fluid from rheumatoid Patients.FEBSL et,1994,35O:9-12.
[24]M Rossignol, A Leclerc, FAAllaert, et al.Primary osteoarthritis of hip knee, and hand in relation to occupational exposure[J].Occup Environ Med,2005,62:772-777'.
[25]查建林.膝骨关节炎的病因与生物力学的关系[J].医药论坛,2009,(1)：35-36.
[26]Oliveria SA, Caffrey M, Linden SM, et al.The analysis of epidemiology about human arthritis[J].Epidemiology,2004,15:326-331.
[27]王序全,李起鸿,机械应力异常与关节软骨退变[J].国外医学生物医学工程分册,1999,22(2)：100-192.
[28]吕存贤,王维佳.膝骨关节炎病因病理研究及中医治疗概况[J].浙江中医学院学报,2005,29(1)：82.
[29]李忆农.细胞因子与关节炎[J].中华风湿病学杂志,2000,24(1)：56-58.
[30]Neidel J,sihulze M,SoveL,et at.Practical significance of cytokine determi-nation in joint Fluid patients with arthroses or rheu-matoid arthritis[J].Z Orthop Ihre Grenzgeb, 1996,(134):381.
[31]顾迁,戴戎,裘世静等.异常高应力导致关节软骨退变机理的形态学研究[J].中华外科杂志,1995,33(10)：597-600.
[32,33]王序全,李起鸿.机械应力异常与关节软骨退变[J].国外医学生物医学工程分册1999,22(2)：100-192.
[34]高翔.益气化瘀利水方干预兔膝骨关节炎软骨中胰岛素样生长因子1的变化[J].中国临床康复,2006,10(39)：95-97.
[35]Lauder SN,Carty SM,Carpenter CE, et al. Interleukinlbeta induced activation of nuclear factor kappab can be inhibited by no el pharmacological agents in osteoarthritis. Rheumatology(Oxford),2007,46(5):752-758.
[36]Sakimura K, Matsumoto T, Miyamoto C, et al.Effects of insulin like growth fact or I on transforming growth factor betal induced chondrogenesis of synovium derived mesenchymal stem cells cultrued in a polyglycolic acid scaffold. Celles T issues Organs,2006,183 (2):55-61.
[37]Blaney Davidson EN, van der Kraan PM,van den Berg WB. TGF beta and osteoarthritis. Osteoarthritis Cartilage,2007,15(6):597-604.
[38]Blaney Davidson EN, Scharstuhl A, Vitters EL, eal. Reduced transforming growth factor-beta signaling in cartilage of old mice:role in impaired repair capacity. Arthritis Res T her,2005,7 (6):R1338-R1347.
[39]van Beuningen HM, van der Kraan PM, Arntz OJ,et al. Transforming growth factor-beta 1 stimulate articular chondrocyte proteoglycan synthesisand induces o steophyte formation in the murine knee joint Lab Invest,1994,71(2):279-290.
[40]Malemud CJ, Killeen W, H ering T M, et a 1. Enhanced sulfated proteogly can core protein synthesis by incubation of rabbit chondrocytes with recombinant transforming growth factor bet a l. J Cell Physiol,1991,149(1):152-159.
[41]Iwanaga H, M atsumoto T, Enomoto H, et al. Enhanced expression of insulin like growth factor binding proteins in human osteoarthritic cartilage detected by immunohistochemistry and in situ hybridization.Osteoarthritis Cartilage,2005, 13(5):439-448.
[42]Barta E, M aroudas A. A theoretical study of the distribution of insulin like g rowth factor in human articular cartilage. J T heor Biol,2006,241(3):628-638.
[43]张德辉,薛刚,黄昌林.应用玻璃酸钠对关节镜术后膝骨关节炎功能恢复的影响[J].中国临床康复,2002,12(6)：1730-1731.
[44]乔聚义,李秋莉,席彬等.低场强MRI对椎体终板骨软骨炎的诊断价值[J].中华实用诊断与治疗杂志.2010,24(11)：1103-1104.
[45]Jasin H E. Autoantibody specificities of immune complexes sequestered in articular cartilage of patients with rheumatoid arthritis and osteoarthritis.Arthritis Rheum,1985, 18:241.
[46]Herman JH, Appel AM.Immunologic modulation of cartilage metabolism.In:Pelletier JP.Edited Proteases:The or involvement in OA. J Rheumatol,1987,14,(Supp 1)1483.
[47]王新文,张民,卫小春.膝骨关节炎发病的免疫机制研究进展[J].中国现代医生,2010,48(18)：16-18.
[48]Burnett BP, Levy R, Cole BJ. Metabolic mechanisms in the pathogenesis of osteoarthritis.[J].Knee Surg,2006;19(3):191-197.
[49]Janet M, Anderson—MacKenzie, Helen—L et al.Fundamental subchondral bone changes in spontaneous knee osteoarthritis[J].The international Journal of Biochemistry& Cell biology,2004,(10):1-13.
[50]李强,张柳.骨性关节炎中软骨下骨与骨退变的关系研究进展[J].中国修复重建外科杂志,2009,23(2)：245-247.
[1]徐叔云,卞如濂,陈修.药理实验方法学.第3版.[M].北京：人民卫生出版社,2002：926.
[2]郭建刚等.骨性关节炎软骨的生化及代谢变化[J].中华骨伤杂志,1998,11(5)：73-74.
[3]李茂强,史晨辉,王永明.骨关节炎的实验研究现况与进展[J].农垦医学2004,26(1)：29.
[4]Aigner T, Sachse A, Gebhard PM, et al.Osteoarthritis:Patho-biology-targets and ways for therapeutic intervention[J]. Adv Drug Delivery Rev,2006,58 (2):128-149.
[5]Gold ring MB. Update on the biology of the chondrocyte and new approaches to treating cartilage diseases[J].Best Pract Res Clin Rheumatol,2006,20(5):1003-1025.
[6]Mar oudasA. Balance betweens welling pressure and collagen tension in and normal degenerate cartilage [J]. Nature,1976,260(5554):808-809.
[7]Fosang AJ, Rogers on FM, East CJ, et al. ADAMTS2 5:The story so far[J]. Eur Cell Mater,2008,15(1):11-26.
[8]PrattaMA, YaoWQ, Decicco C, et al. Aggrecan protects cartilage collagen from proteolytic cleavage[J]. J Biol Chem,2003,278(46):45539-45545.
[9]Nagase H, Woessner JF Jr. Matrix metallopromteinases [J]. J Boil Chem 1999,274(3):21491-21494.
[10]Yu LP Jr. Smith GN Jr.HastykA.et al.Doxycycllne inhibits type II Collagen olytic activity of extracts from human osteoarthritic Cartilage and of gelatinase.J Rheumatol,1991,18(10):1450-1452..
[11]Murphy G, Knauper V, Atkinson, et al. Matrix metalloproteinases in arthritic disease.Arthriistes,2002,4(Suppl 3):39-49.
[12]Lijnen HR.Elements of the fibrinolytic system[J].Ann NY Acad Sci,2001,936:226-36.
[13]徐海涛,陈游,陈立科,不同干预因素对膝骨关节炎患者滑液中MMP-3和TIMP-1的影响[J].中南大学学报(医学版),2008,33(1)47.
[14]Taylor DJ, Cheung NT, Dawes PT. Increased serum pro-MMP-3 in inflammatory arthritides:a potential indicator of synovial inflammatory monokie activity[J].Annn Rheum Dis,1994,53(11):768-772.
[15]Ichikawa Y, Yamada C,Hori ki T,et al.Serum matrix metalloproteinse-3 and fibrin degradation product levels correlate with clinical disease activity in rheumatoid arthritis[J].Clin Exp Rheumatol,1998,16 (5):533-54.
[16]王晓霞,李小峰,王来远等.类风湿关节炎血清基质金酶-3水平及其影响因素的探讨[J].中华风湿病学杂志,(1)：16-19.
[17]魏平,付爽,王俊祥.基质金属蛋白酶3在类风湿关节炎达及其意义[J].临床荟萃,2006,21(19)：1389-1390.
[18]朱宝玉,陈琼,王万春.基质金属蛋白酶-3(MMP-3)在创伤后关节软骨退变中的作用[J].实用骨科杂志.2006,12(1)：35.
[19]陈希,涂长青,刘丽娟.补肾活血方对膝骨关节炎蛋白聚糖和Ⅱ型胶原表达的影响[J].新中医,2008,40(8)：47.
[20]梁祖建,韩清民,张还添.补肾活血方对骨关节炎关节软骨保护的效应与机制[J].江苏中医药,2009,41(3)：34.
[21]江艳,李荣亨.复元胶囊对膝骨关节炎患者血清MMP-3和TIMP-1表达变化的影响[J].西南国防医药,2010,20(9)：934.
[22]李超等.蛋白聚糖酶和骨关节炎.[J].北京大学学报(医学版),2009,41：611-613.
[23]Sh arif M, Kir wan J, Cham iN, et al A 5-yr longitudinal study of type Ⅱ A collagen synthesis and total type Ⅱ collagen degradation in patients with knee osteoarthritis-association with disease progression [J]. Rheumatology,2007,46:938-943.
[24]Sh inj iU, Yos h i h it o N, Yas um asa S, et a.l Fluid Levels of Chon-droit in Sulfate in Patients with Osteoarthritis of the Knee Joint[J].JN ppon Med Sch,2001,68:165-170.
[1]刘向前.中西医结合治疗膝关节骨关节炎(第1版)[M].人民军医出版社,2011：23.
[2-3]刘献祥,林燕萍.中西医结合治疗膝关节骨关节炎(第1版)[M].人民卫生出版社,2009：3.
[4]张蓉,李峰,王常海.经筋理论在膝关节骨性关节炎发病机制及治疗中的作用[J].中国康复医学杂志,2007,22(7)：644.
[5]任平霞.综合治疗早中期膝关节骨性关节炎169例[J].光明中医,2011,26(8)：1631-1632.
[6]Videman T. Experimental osteoarthritis in the rabbit:comparison of different periods of repeated immobilization [J].Acta Orthop Scand,1982,53(3):339-347.
[7]Salo PT, Hogervorst T, Seerattan RA. Selective joint denervation promotes knee osteoarthritis in the aging rat [J].J orthop Res,2002,20(6):1256-1264.
[8]Yamamoto K Yamamoto K, Shishido T, Masaoka T, et al. Morphologicalstudies on the ageing and osteoarthritis of the articular cartilage in C57 black mice [J]. Orthop Surg (kong Kong),2005,13(1):8-18.
[9]刘伯龄,刘献祥.骨性关节炎动物模型的建立概况[J].福建中医药,2003,34(4)：49.
[10]杨峰,史索道.用木瓜蛋白酶建立兔颚须关节骨关节炎模型[J].华西口腔医学杂志,2002,12(5)：330-333.
[11]Kikuchi T, Sakuta T, Yamaguchi T. Intra-articular injection of collagenase induces experimental osteoarthritis in mature rabbits[J].Osteoarthritis Cartilage, 1998,6(3):177-186.
[12]T sai CL,L iu TK. Estradiol-induced knee osteoarthrosis in ovariectomized rabbits[J].Clin Orthop,1993,(291):295-302.
[13]Sakano Y, Terada N,U eda H, et al. Histo logical study of articular cartilage in experimental rat knee arthritis induced by intracapsular injection of cationic poly ethleneimine[J].Med Electron Micro sc,2000,33(4):246-257.
[14]膝关节骨关节炎动物模型的构建[J].中国中医骨伤科杂志,2012,20(12)：71.
[15]M cDevitt CA, Gilbertson EMM. M uir H. An experimental model of osteoarthritis. Early morphological and biochemical changes[J].J bone Joint Surg,1977,59.(B):24-30.
[16]顾廷,戴克戎,裘世静等.应力降低导致关节软骨退变机理形态学研究[J].中华骨科杂志,1995,15(9)：631-633.
[18]M arijnissen AC, Van Roermund PM, Te Koppele JM, et al. The canine'groove'model, compared with the ACL T model of osteoarthritis[J].Osteoarthritis Cartilage,2002,10(2):145-155.
[19]白希壮,任继尧.选择性臀肌切断诱发骨关节炎实验模型[J].中华骨科杂志,1994,14(2)：118-120.
[20]刘蜀彬,孔祥星,史迎昌等.股骨干骺端髓内血运阻断诱发兔膝关节骨性关节炎模型[J].中国矫形外科杂志,2003,11(24)：1706-1707.
[21]胡波,胡志俊,张有为等.骨关节炎动物模型：人工诱导与动物自发[J].中国组织工程研究与临床康复,2010,14(33)：6202-6205.
[22]刘媛,赵驻军,王红芹等.寒冷刺激鸡膝关节软骨显微结构变化观察[J].河北农业大学学报,2008,31(2)：93-95.
[23]陆志华,姚伟武,胡曙光等应用磁共振生理学成像定量测定兔膝关节软骨退变的研究[J].医学影像学杂志,2009,19(6)：739-744.
[24]王学谦,娄思权,侯筱魁等译.骨关节炎诊断与治疗(第四版)[M].天津科技翻译出版公司,2008,3：59-77.
[25]邱宜兴兔关节制动引起关节软骨退变作用的实验研究[J].中华骨科杂志.1987；25(3)：175.
[26]吴海山,周维江,张中南.老年性退变膝[J].中华老年医学杂志,1994,13(6)：330.
[27]吴毅,范振华,屠丹云等.运动促进运动后兔膝关节组织修复的应用[J].中国康复医学杂志,1993,8(1)：17-20.
[28]祖启明,王庆良,郭正成.关节持续被动运动的基础研究和临床应用[J].国外医学物理医学与康复学分册,1995,15(1)：155-158.
[29]邱贵兴,王桂生.兔膝关节制动引起关节软骨退变的实验研究[J].中华外科杂志,1987,25(30)：175-177.
[1]Lequesne MG, Samsons M. Indices of severity in osteoarthritis of weight bearing joints [J].JRheumatol Supp,1991,27:16-18.
[2]Lequesne MG, Mery C, Samson M, et al. Comparison between the WOMAC and the Lequesne indices in patients with knee and hip osteoarthritis [J].Osteoarthritis Cartilage, 1998,6(6):441-442.
[3]Lequesne MG, Maheu E. Methodology of clinical trials in hand osteoarthritis: conventional and proposed tools [J]. Osteoarthritis Cartilage,2000,8(Suppl A):64-69.
[4]Lequesne M, Maheu E, Cadet C, et al. Structural effect of avocado/soybean unsaponifiables on joint space loss in osteoarthritis of the hip [J]. Arthritis Rheum,2002, 47(1):50-58.
[5]Lequesne MG, Maheu E. Clinical and radiological evaluation of hip, knee and hand osteoarthritis [J]. Aging Clin Exp Res,2003,15(5):380-390.
[6]龚权,程传国,程龙.药物电针刀疗法对兔膝骨性关节炎软骨细胞凋亡的影响[J].中国中西医结合杂志基础理论研究特集,2004,24：195-196.
[7]曾贵刚,张秀芬,权伍成。针刀松解法对膝骨性关节炎应力刺激及临床症状的影响[J].中华中医药学刊,2009,27(1)：66-68.
[8]McCarry DJ:Arthritis and Allied Condition1Oth ed, Lea and Febiger, PhiladelPhia,1985.
[9]Doyle DV:J pathol,1982,136(3):199.
[10]BurgerC:Acta Orthop Scand,1983,54(1):132.
[11]Egan MC:Arhritis Rheum,1982,25(2):204.
[12]InoueH:Int Orthop,1981,5(1):47.
[13]Finsterbush A:Clin Orthop,1977,111:290.
[14]Palmoski M:Arthritis Rheum.1981,24(10):1329.
[15]Fabry G:Clin Orthop,1980,153:253.
[16]Radin EL:J Biomech,1973,6(1):51.
[17]Sapolsky AI:Arthritis Rheum.1982,25(8):981.
[18]Chadilly FN:Fine Structure of Synovial Joints Butterworth,London,1983.
[19]朱汉章主编.针刀医学.第1版.[M].北京：中国中医药出版社,2004：128.
[1]徐叔云,卞如濂,陈修.药理实验方法学[M].3版.北京：人民卫生出版社,2002：926.
[2]Aigner T, Sachse A, Gebhard PM, et al. Osteoarthritis:Patho-biology-targets and ways for therapeutic intervention[J]. Adv Drug Delivery Rev,2006,58 (2):128-149.
[3]Gold ringMB. Update on the biology of the chondrocyte and new approaches to treating cartilage diseases[J]. Best Pract Res Clin Rheumatol,2006,20 (5):1003-1025.
[4]Mar oudasA. Balance bet ween s welling p ressure and collagen tension in normal and degenerate cartilage [J]. Nature,1976,260(5554):808-809.
[5]Fosang AJ, Rogers on FM, East CJ, et al. ADAMTS2 5:The story so far[J]. Eur Cell Mater,2008,15(1):11-26.
[6]PrattaMA, YaoWQ, Decicco C, et al. Aggrecan p rotects cartilagecollagen from pr oteolytic cleavage [J]. J Biol Chem,2003,278(46):45539-45545.
[7]Nagase H, Woessner JF Jr. Matrix metallopromteinases [J]. J Boil Chem,1999,274(3): 21491-21494.
[8]Yu LP Jr. SmIth GN Jr.HastykA.et al.Doxycycllne inhibits type Ⅱ Collagenolytic activity of extracts fromhuman osteoarthritic Cartilage and of gelatinase.J Rheumatol,1991, 18(10):1450-1452.。
[9]Murphy G, Knauper V, Atkinson, et al. Matrix metalloproteinases in arthritic disease. Arthriistes,2002,4(Su ppl 3):39-49.
[10]LijnenHR.Elements of the fibrinolytic system[J].Ann NY Acad Sci,2001,936:226-36.
[11]徐海涛,陈游,陈立科.不同干预因素对膝骨关节炎患者滑液中MMP-3和TIMP-1的影响[J].中南大学学报(医学版),2008,33(1)：47.
[12]庞坚,曹月龙,詹红生.骨关节炎的“筋”“骨”之辨[J].上海中医药大学学报,2012,26(1).
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