左归丸防治骨质疏松症的动物模型和评价方法
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摘要
左归丸是滋阴补肾、填精益髓的经典名方,体现了"阳中求阴"的配伍思路,是防治骨质疏松症(OP)的常用有效方剂之一。OP动物模型模拟了OP在人体的病理状态和发病机制,是OP防治和新药研发的重要手段。该文详细论述了左归丸防治OP的2大类常用动物模型和特点,即原发性OP动物模型,包括去卵巢法动物模型和自发性老年OP动物模型;继发性OP动物模型,包括糖皮质激素诱导性动物模型,环磷酰胺诱导性动物模型以及肾大部切除性动物模型。左归丸防治OP的效用评价,包括酶联免疫吸附测定实验检测骨吸收标记物和骨形成标记物含量,双能X射线检测骨密度,显微CT分析骨小梁数量、骨小梁分离度、骨小梁厚度、骨体积/组织体积比以及骨表面/体积比,以衡量骨微结构水平,苏木精-伊红染色法观察骨病理形态,生物力学实验测试骨最大载荷力等生物力学特性。OP动物模型的成功与否,以及左归丸的药效作用,必须与5种不同的客观评价方法结合起来进行综合衡量,才能为左归丸的基础和临床研究提供科学的参考依据。但OP动物模型有待进一步优化,以凸显左归丸治疗OP的中医病机和证候特征。
Zuoguiwan is a classic prescription for replenishing vital essence,tonifying kidney-Yin and nourishing the bone marrow. Zuoguiwan is one of the effective prescriptions for the prevention and treatment of osteoporosis( OP),which reflects the thought of Reinforcing Yang from Yin. The OP animal model simulates the pathological state and pathogenesis of OP in human,which is an important means to research the pathogenesis of OP and verify the effect of drugs. In this paper,two kinds of animal models and characteristics of Zuoguiwan in the prevention and treatment of osteoporosis were discussed in details. They are the primary osteoporosis animal models,including ovariectomized animal models and spontaneous elderly osteoporosis animal models,and the secondary osteoporosis models,including glucocorticoid-induced animal models,cyclophosphamide-induced animal models and subtotal nephrectomy animal models. The evaluation methods of Zuoguiwan in preventing and treating OP,including bone absorption markers and bone formation markers analyzed by enzyme-linked immunosorbent assay( ELISA),bone mineral density detected with dual-energy X-ray,the number of trabeculae,trabecular segregation,trabecular thickness,bone volume/tissue volume ratio and bone surface/volume ratio analyzed using micro-CT,bone pathological morphology observed by hematoxylin-eosin staining,bone biomechanical properties,such as the maximum load force based on biomechanical test. In order to provide scientific reference for the basic and clinical research of Zuoguiwan,the OP animal models and the pharmacodynamic effect of Zuoguiwan are evaluated comprehensively with five different and objective evaluation methods. However,the animal model of OP needs to be further optimized to highlight the pathogenesis and syndrome characteristics of Zuoguiwan in the treatment of OP.
Zuoguiwan is a classic prescription for replenishing vital essence,tonifying kidney-Yin and nourishing the bone marrow. Zuoguiwan is one of the effective prescriptions for the prevention and treatment of osteoporosis( OP),which reflects the thought of Reinforcing Yang from Yin. The OP animal model simulates the pathological state and pathogenesis of OP in human,which is an important means to research the pathogenesis of OP and verify the effect of drugs. In this paper,two kinds of animal models and characteristics of Zuoguiwan in the prevention and treatment of osteoporosis were discussed in details. They are the primary osteoporosis animal models,including ovariectomized animal models and spontaneous elderly osteoporosis animal models,and the secondary osteoporosis models,including glucocorticoid-induced animal models,cyclophosphamide-induced animal models and subtotal nephrectomy animal models. The evaluation methods of Zuoguiwan in preventing and treating OP,including bone absorption markers and bone formation markers analyzed by enzyme-linked immunosorbent assay( ELISA),bone mineral density detected with dual-energy X-ray,the number of trabeculae,trabecular segregation,trabecular thickness,bone volume/tissue volume ratio and bone surface/volume ratio analyzed using micro-CT,bone pathological morphology observed by hematoxylin-eosin staining,bone biomechanical properties,such as the maximum load force based on biomechanical test. In order to provide scientific reference for the basic and clinical research of Zuoguiwan,the OP animal models and the pharmacodynamic effect of Zuoguiwan are evaluated comprehensively with five different and objective evaluation methods. However,the animal model of OP needs to be further optimized to highlight the pathogenesis and syndrome characteristics of Zuoguiwan in the treatment of OP.
引文
[1] Rossini M,Adami S,Bertoldo F,et al. Guidelines for the diagnosis,prevention and management of osteoporosis[J]. Reumatismo,2016,68(1):1-39.
[2] Muhammad A, Mada S B, Malami I, et al.Postmenopausal osteoporosis and breast cancer:The biochemical links and beneficial effects of functional foods[J]. Biomed Pharmacother,2018,107:571-582.
[3] Ichimaru R, Tominari T, Yoshinouchi S, et al.Raloxifene reduces the risk of local alveolar bone destruction in a mouse model of periodontitis combined with systemic postmenopausal osteoporosis[J]. Arch Oral Biol,2018,85:98-103.
[4] YU F, XIA W. The epidemiology of osteoporosis,associated fragility fractures,and management gap in China[J]. Arch Osteoporos,2019,14(1):32.
[5] PENG C,LI Z,HU Y. Prevalence of osteoporosis in China:a Meta-analysis and systematic review[J]. BMC Public Health,2016,16(1):1039-1050.
[6] CHEN H,WU M,KUBO K Y. Combined treatment with a traditional Chinese medicine, Hachimi-jio-gan(Ba-Wei-Di-Huang-Wan)and alendronate improves bone microstructure in ovariectomized rats[J]. J Ethnopharmacol,2012,142(1):80-85.
[7] Black D M,Greenspan S L,Ensrud K E,et al. The effects of parathyroid hormone and alendronate alone or in combination in postmenopausal osteoporosis[J]. N Engl J Med,2003,349(13):1207-1215.
[8] CHENG M,WANG Q,FAN Y,et al. A traditional Chinese herbal preparation, Er-Zhi-Wan, prevent ovariectomy-induced osteoporosis in rats[J]. J Ethnopharmacol,2011,138(2):279-285.
[9]张景岳.景岳全书[M].李继明,王大淳整理.北京:中国中医药出版社,2011:1283-1284.
[10]谭峰,樊巧玲,卞玉群,等.左、右归丸对去卵巢骨质疏松症大鼠模型骨密度及骨代谢的影响[J].中国实验方剂学杂志,2015,21(9):137-140.
[11]李然,黄瀚,刘立萍,等.左归丸对去卵巢大鼠骨组织降钙素受体蛋白表达的影响[J].中华中医药学刊,2018,36(12):2993-2995.
[12]谭彪,孙瑗,杨世鹏.左归丸加减内服配合玻璃酸钠关节内注射治疗肝肾亏虚型膝骨关节炎近期疗效观察[J].中国社区医师,2018,34(22):81-82.
[13] Minne H W,Pfeifer M,Hinz C. Osteoporosis--mode or disease[J]. Med Klin:Munich, 2004,99(11):663-670.
[14]郭鱼波,马如风,王丽丽,等.骨质疏松动物模型及其评价方法的研究进展[J].中国骨质疏松杂志,2015,21(9):1149-1154.
[15]鞠大宏,张春英,王安民,等.左归丸对去卵巢所致大鼠骨质疏松症的治疗作用[J].中国中医基础医学杂志,2001,7(3):17-20.
[16]黄宏兴,柴生颋,黄红,等.骨质疏松症中医证型的聚类分析[J].广州中医药大学学报,2007,24(3):180-183,187.
[17] Chow S K,Leung K S,QIN J,et al. Mechanical stimulation enhanced estrogen receptor expression and callus formation in diaphyseal long bone fracture healing in ovariectomy-induced osteoporotic rats[J]. Osteoporos Int,2016,27(10):2989-3000.
[18] LIU M J,LI Y,PAN J H,et al. Effects of Zuogui pill(左归丸)on Wnt singal transduction in rats with glucocorticoid-induced osteoporosis[J]. J Tradit Chin Med,2011,31(2):98-102.
[19] LIU F,TAN F,TONG W,et al. Effect of Zuoguiwan on osteoporosis in ovariectomized rats through RANKL/OPG pathway mediated byβ2AR[J]. Biomed Pharmacoth,2018,103:1052-1060.
[20]付小卫.基于破骨细胞和成骨细胞调控通路探讨左、右归丸对不同性别去势大鼠骨质疏松症的作用差异及其机制[D].北京:中国中医科学院,2014.
[21] Jilka R L,Weinstein R S,Takahashi K,et al. Linkage of decreased bone mass with impaired osteoblastogenesis in a murine model of accelerated senescence[J]. J Clin Invest,1996,97(7):1732-1740.
[22] Azuma K,ZHOU Q,Kubo K Y. Morphological and molecular characterization of the senile osteoporosis in senescence-accelerated mouse prone 6(SAMP6)[J].Med Mol Morphol,2018,51(3):139-146.
[23] CHEN H, Shoumura S, Emura S. Ultrastructural changes in bones of the senescence-accelerated mouse(SAMP6):a murine model for senile osteoporosis[J].Histol Histopathol,2004,19:677-685.
[24]李琳.衰老海马Notch、Wnt和MAPK信号通路的关键蛋白表达变化及补肾方作用机理研究[D].郑州:河南中医药大学,2017.
[25]刘梅洁,鞠大宏,赵宏艳,等.“肾主骨”的机理研究—左归丸含药血清对破骨细胞分化调控因子OPG、RANKL蛋白表达的影响[J].中国中医基础医学杂志,2009,15(3):184-187,196.
[26] WANG H,SHAN X B,QIAO Y J. PDK2 promotes chondrogenic differentiation of mesenchymal stem cells by upregulation of Sox6 and activation of JNK/MAPK/ERK pathway[J]. Braz J Med Biol Res,2017,50(2):e5988.
[27] HU Y,CHAN E,WANG S X,et al. Activation of p38mitogen-activated protein kinase is required for osteoblast differentiation[J]. Endocrinology,2003,144(5):2068-2074.
[28]刘立萍,任艳玲,李然,等.左归丸含药血清通过p38MAPK信号通路诱导MC3T3-E1成骨细胞的分化[J].中国老年学杂志,2013,33(7):1579-1581.
[29]刘立萍,任艳玲,李然,等. JNK通路对左归丸含药血清调控MC3T3成骨细胞Runx2 mRNA表达的影响[J].中国实验方剂学杂志,2012,18(14):143-146.
[30] ZHANG Z,REN H,SHEN G,et al. Animal models for glucocorticoid-induced postmenopausal osteoporosis:an updated review[J]. Biomed Pharmacother,2016,84:438-446.
[31] ZHAO J,LI Y,ZHANG H,et al. Preventative effects of metformin on glucocorticoid-induced osteoporosis in rats[J]. J Bone Miner Metab,2019,doi:10. 1007/S00774-019-00989-y.
[32] DING Z,SHI H,YANG W. Osteoprotective effect of cimiracemate in glucocorticoid-induced osteoporosis by osteoprotegerin/receptor activator of nuclear factorκB/receptor activator of nuclear factor kappa-B ligand signaling[J]. Pharmacology, 2019,103(3/4):163-172.
[33] LIU Y,ZUO G,MENG X,et al. Adrenomedullin inhibits osteoclast differentiation through the suppression of receptor activator of nuclear factor-κB ligand-induced nuclear factor-κB activation in glucocorticoid-induced osteoporosis[J]. Exp Ther Med, 2017,14(5):4009-4016.
[34] Wolski H,Drews K,Bogacz A,et al. The RANKL/RANK/OPG signal trail:significance of genetic polymorphisms in the etiology of postmenopausal osteoporosis[J]. Ginekol Pol,2016,87(5):347-352.
[35]刘梅洁,潘静华,李艳,等.左归丸对糖皮质激素所致骨质疏松大鼠血清中BGP,IGF-Ⅰ含量的影响[J].中国实验方剂学杂志,2011,17(16):133-136.
[36]张志达,任辉,沈耿杨,等.左归丸调控DKK1靶点防治糖皮质激素性骨质疏松[J].中国组织工程研究,2018,22(16):2520-2525.
[37] Saarto T,Blomqvist C,Valimaki M,et al. Chemical castration induced by adjuvant cyclophosphamide,methotrexate, and fluorouracil chemotherapy causes rapid bone loss that is reduced by clodronate:a randomized study in premenopausal breast cancer patients[J]. J Clin Oncol,1997,15(4):1341-1347.
[38] Murphy C, Muscat A, Ashley D, et al. Tailored NEOadjuvant epirubicin, cyclophosphamide and nanoparticle albumin-bound paclitaxel for breast cancer:The phaseⅡNEONAB trial-clinical outcomes and molecular determinants of response[J]. PLo S One,2019,14(2):e0210891.
[39] Sugita J,Kagaya Y,Miyamoto T,et al. Myeloablative and reduced-intensity conditioning in HLAhaploidentical peripheral blood stem cell transplantation using post-transplant cyclophosphamide[J]. Bone Marrow Transplant,2019,54(3):432-441.
[40] PEI Y,ZHENG K,SHANG G,et al. Therapeutic effect of strontium ranelate on bone in chemotherapy-induced osteopenic rats via increased bone volume and reduced bone loss[J]. Biol Trace Elem Res,2019,187(2):472-481.
[41] ZHAO D, WANG C, ZHAO Y, et al.Cyclophosphamide causes osteoporosis in C57BL/6 male mice:suppressive effects of cyclophosphamide on osteoblastogenesis and osteoclastogenesis[J].Oncotarget,2017,8(58):98163-98183.
[42]刘政.环磷酰胺致骨质疏松大鼠模型的建立及左归丸的防治作用[D].广州:暨南大学,2015.
[43]彭慧娟,朱玲,许丽绵,等.左归丸对环磷酰胺卵巢早衰大鼠卵巢Bax、Bcl-2及Cyt-c的影响[C]//中国中西医结合学会妇产科专业委员会.第9届中国中西医结合学会妇产科专业委员会第二次学术会议论文集.昆明,2017:8-10.
[44]周家俊,关鑫,赵东.部分肾切除加高磷饮食所致大鼠肾性骨病模型研究[J].中国中西医结合肾病杂志,2002(10):567-569.
[45]袁军,王小琴,马晓红,等.左归丸对5/6肾大部切除模型并肾性骨病大鼠甲状旁腺的影响[J].中医药学报,2013,41(1):76-79.
[46]王小琴,袁军,马晓红,等.左归丸对5/6肾大部切除模型并肾性骨病大鼠骨代谢指标的影响[J].中国中西医结合肾病杂志,2012,13(7):584-586.
[47] LONG F. Building strong bones:molecular regulation of the osteoblast lineage[J]. Nat Rev Mol Cell Biol,2011,13(1):27-38.
[48] Gomes A M,Douglas-de-Oliveira D W,Ferreira S D,et al. Periodontal disease,peri-implant disease and levels of salivary biomarkers IL-1β, IL-10, RANK, OPG,MMP-2,TGF-βand TNF-α:follow-up over 5 years[J].J Appl Oral Sci,2019,27:e20180316.
[49] LI B,Boast S,Kenia de los S,et al. Mice deficient in Abl are osteoporotic and have defects in osteoblast maturation[J]. Nat Genet,2000,24(3):304-308.
[50] Burshell A L,Mricke R,Correa-Rotter R,et al.Correlations between biochemical markers of bone turnover and bone density responses in patients with glucocorticoid-induced osteoporosis treated with teriparatide or alendronate[J]. Bone,2010,46(4):935-939.
[51] Glover S J,Eastell R,McCloskey E V,et al. Rapid and robust response of biochemical markers of bone formation to teriparatide therapy[J]. Bone,2009,45(6):1053-1058.
[52] Akbari Moghaddam Kakhki R,LU Z,Thanabalan A,et al. Eimeria challenge adversely affected long bone attributes linked to increased resorption in 14-day-old broiler chickens[J]. Poult Sci, 2019,98(4):1615-1621.
[53] Kim S K,Lee M H,Rhee M H. Studies on the effects of biomedicinal agents on serum concentration of Ca2+,P and ALP activity in osteoporosis-induced rats[J]. J Vet Sci,2003,4(2):151-154.
[54] YAO H H,TANG S M,WANG Z M,et al. Study of bone mineral density and serum bone turnover markers in newly diagnosed systemic lupus erythematosus patients[J]. J Peking Univ Health Sci,2018,50(6):998-1003.
[55] Hatate K,Kayano M,Kawashima C,et al. The effect of colostrum intake on osteoprotegerin and bone metabolic markers in the blood of newborn calves during the first week of life[J]. J Vet Med Sci,2019,81(1):15-21.
[56] PAN C,LIU X,LI T,et al. Kinetic of bone turnover markers after osteoporotic vertebral compression fractures in postmenopausal female[J]. J Orthop Surg Res,2018,13(1):314.
[57] Rief H,Omlor G,Akbar M,et al. Biochemical markers of bone turnover in patients with spinal metastases after resistance training under radiotherapy-a randomized trial[J]. BMC Cancer,2016,16:231.
[58]叶素敏,谭峰,刘飞祥,等.左归丸对去卵巢大鼠下丘脑Orexin及其受体mRNA表达的影响[J].中国实验方剂学杂志,2019, doi:org/10. 13422/j. cnki.syfjx. 20191021.
[59]童伟伟.基于补肾益癸理论探讨左归丸防治PMOP的疗效机制[D].南京:南京中医药大学,2018.
[60]卢建华,王维佳,储小兵,等.补肾方对去卵巢大鼠Ⅰ型胶原代谢的影响[J].中华中医药杂志,2010,25(7):1109-1111.
[61]孙月娇,任艳玲,曲宁宁,等.左、右归丸及其拆方对去卵巢骨质疏松模型大鼠的比较研究[J].中国骨质疏松杂志,2015,21(8):910-913,921.
[62]李泽钊,邓建强.地仲强骨胶囊治疗肾虚兼气血不足型骨质疏松症的疗效及对骨代谢、骨转化的影响[J].中国实验方剂学杂志,2018,24(8):159-164.
[63] Kanis J A,Cooper C,Rizzoli R,et al. Review of the guideline of the American College of Physicians on the treatment of osteoporosis[J]. Osteoporos Int,2018,29(7):1505-1510.
[64] Hanson N A, Bagi C M. Alternative approach to assessment of bone quality using micro-computed tomography[J]. Bone,2004,35(1):326-333.
[65]刘晓,于文涛,靳贺超,等.补肾通络方对肺纤维化大鼠肺组织TGF-β1/Smads通路的影响[J].中国实验方剂学杂志,2019,25(2):96-102.
[66]卢建华,王维佳,陈明涛,等.补肾方对去卵巢大鼠腰椎骨形态计量学的影响[J].中医正骨,2011,23(5):24-26.
[67]王如然,冯艳华,张玉杰,等.左归丸、右归丸对去卵巢骨质疏松模型大鼠的效用[J].基因组学与应用生物学,2018,37(3):1104-1109.
[68] Massie A M,Kapatkin A S,Garcia T C,et al. Effects of hole diameter on torsional mechanical properties of the rabbit femur[J]. Vet Comp Orthop Traumatol,2019,32(1):51-58.
[69]董苗淼.运动配合左归丸对去卵巢大鼠骨生物力学及相关激素的影响[D].上海:上海体育学院,2010.
[2] Muhammad A, Mada S B, Malami I, et al.Postmenopausal osteoporosis and breast cancer:The biochemical links and beneficial effects of functional foods[J]. Biomed Pharmacother,2018,107:571-582.
[3] Ichimaru R, Tominari T, Yoshinouchi S, et al.Raloxifene reduces the risk of local alveolar bone destruction in a mouse model of periodontitis combined with systemic postmenopausal osteoporosis[J]. Arch Oral Biol,2018,85:98-103.
[4] YU F, XIA W. The epidemiology of osteoporosis,associated fragility fractures,and management gap in China[J]. Arch Osteoporos,2019,14(1):32.
[5] PENG C,LI Z,HU Y. Prevalence of osteoporosis in China:a Meta-analysis and systematic review[J]. BMC Public Health,2016,16(1):1039-1050.
[6] CHEN H,WU M,KUBO K Y. Combined treatment with a traditional Chinese medicine, Hachimi-jio-gan(Ba-Wei-Di-Huang-Wan)and alendronate improves bone microstructure in ovariectomized rats[J]. J Ethnopharmacol,2012,142(1):80-85.
[7] Black D M,Greenspan S L,Ensrud K E,et al. The effects of parathyroid hormone and alendronate alone or in combination in postmenopausal osteoporosis[J]. N Engl J Med,2003,349(13):1207-1215.
[8] CHENG M,WANG Q,FAN Y,et al. A traditional Chinese herbal preparation, Er-Zhi-Wan, prevent ovariectomy-induced osteoporosis in rats[J]. J Ethnopharmacol,2011,138(2):279-285.
[9]张景岳.景岳全书[M].李继明,王大淳整理.北京:中国中医药出版社,2011:1283-1284.
[10]谭峰,樊巧玲,卞玉群,等.左、右归丸对去卵巢骨质疏松症大鼠模型骨密度及骨代谢的影响[J].中国实验方剂学杂志,2015,21(9):137-140.
[11]李然,黄瀚,刘立萍,等.左归丸对去卵巢大鼠骨组织降钙素受体蛋白表达的影响[J].中华中医药学刊,2018,36(12):2993-2995.
[12]谭彪,孙瑗,杨世鹏.左归丸加减内服配合玻璃酸钠关节内注射治疗肝肾亏虚型膝骨关节炎近期疗效观察[J].中国社区医师,2018,34(22):81-82.
[13] Minne H W,Pfeifer M,Hinz C. Osteoporosis--mode or disease[J]. Med Klin:Munich, 2004,99(11):663-670.
[14]郭鱼波,马如风,王丽丽,等.骨质疏松动物模型及其评价方法的研究进展[J].中国骨质疏松杂志,2015,21(9):1149-1154.
[15]鞠大宏,张春英,王安民,等.左归丸对去卵巢所致大鼠骨质疏松症的治疗作用[J].中国中医基础医学杂志,2001,7(3):17-20.
[16]黄宏兴,柴生颋,黄红,等.骨质疏松症中医证型的聚类分析[J].广州中医药大学学报,2007,24(3):180-183,187.
[17] Chow S K,Leung K S,QIN J,et al. Mechanical stimulation enhanced estrogen receptor expression and callus formation in diaphyseal long bone fracture healing in ovariectomy-induced osteoporotic rats[J]. Osteoporos Int,2016,27(10):2989-3000.
[18] LIU M J,LI Y,PAN J H,et al. Effects of Zuogui pill(左归丸)on Wnt singal transduction in rats with glucocorticoid-induced osteoporosis[J]. J Tradit Chin Med,2011,31(2):98-102.
[19] LIU F,TAN F,TONG W,et al. Effect of Zuoguiwan on osteoporosis in ovariectomized rats through RANKL/OPG pathway mediated byβ2AR[J]. Biomed Pharmacoth,2018,103:1052-1060.
[20]付小卫.基于破骨细胞和成骨细胞调控通路探讨左、右归丸对不同性别去势大鼠骨质疏松症的作用差异及其机制[D].北京:中国中医科学院,2014.
[21] Jilka R L,Weinstein R S,Takahashi K,et al. Linkage of decreased bone mass with impaired osteoblastogenesis in a murine model of accelerated senescence[J]. J Clin Invest,1996,97(7):1732-1740.
[22] Azuma K,ZHOU Q,Kubo K Y. Morphological and molecular characterization of the senile osteoporosis in senescence-accelerated mouse prone 6(SAMP6)[J].Med Mol Morphol,2018,51(3):139-146.
[23] CHEN H, Shoumura S, Emura S. Ultrastructural changes in bones of the senescence-accelerated mouse(SAMP6):a murine model for senile osteoporosis[J].Histol Histopathol,2004,19:677-685.
[24]李琳.衰老海马Notch、Wnt和MAPK信号通路的关键蛋白表达变化及补肾方作用机理研究[D].郑州:河南中医药大学,2017.
[25]刘梅洁,鞠大宏,赵宏艳,等.“肾主骨”的机理研究—左归丸含药血清对破骨细胞分化调控因子OPG、RANKL蛋白表达的影响[J].中国中医基础医学杂志,2009,15(3):184-187,196.
[26] WANG H,SHAN X B,QIAO Y J. PDK2 promotes chondrogenic differentiation of mesenchymal stem cells by upregulation of Sox6 and activation of JNK/MAPK/ERK pathway[J]. Braz J Med Biol Res,2017,50(2):e5988.
[27] HU Y,CHAN E,WANG S X,et al. Activation of p38mitogen-activated protein kinase is required for osteoblast differentiation[J]. Endocrinology,2003,144(5):2068-2074.
[28]刘立萍,任艳玲,李然,等.左归丸含药血清通过p38MAPK信号通路诱导MC3T3-E1成骨细胞的分化[J].中国老年学杂志,2013,33(7):1579-1581.
[29]刘立萍,任艳玲,李然,等. JNK通路对左归丸含药血清调控MC3T3成骨细胞Runx2 mRNA表达的影响[J].中国实验方剂学杂志,2012,18(14):143-146.
[30] ZHANG Z,REN H,SHEN G,et al. Animal models for glucocorticoid-induced postmenopausal osteoporosis:an updated review[J]. Biomed Pharmacother,2016,84:438-446.
[31] ZHAO J,LI Y,ZHANG H,et al. Preventative effects of metformin on glucocorticoid-induced osteoporosis in rats[J]. J Bone Miner Metab,2019,doi:10. 1007/S00774-019-00989-y.
[32] DING Z,SHI H,YANG W. Osteoprotective effect of cimiracemate in glucocorticoid-induced osteoporosis by osteoprotegerin/receptor activator of nuclear factorκB/receptor activator of nuclear factor kappa-B ligand signaling[J]. Pharmacology, 2019,103(3/4):163-172.
[33] LIU Y,ZUO G,MENG X,et al. Adrenomedullin inhibits osteoclast differentiation through the suppression of receptor activator of nuclear factor-κB ligand-induced nuclear factor-κB activation in glucocorticoid-induced osteoporosis[J]. Exp Ther Med, 2017,14(5):4009-4016.
[34] Wolski H,Drews K,Bogacz A,et al. The RANKL/RANK/OPG signal trail:significance of genetic polymorphisms in the etiology of postmenopausal osteoporosis[J]. Ginekol Pol,2016,87(5):347-352.
[35]刘梅洁,潘静华,李艳,等.左归丸对糖皮质激素所致骨质疏松大鼠血清中BGP,IGF-Ⅰ含量的影响[J].中国实验方剂学杂志,2011,17(16):133-136.
[36]张志达,任辉,沈耿杨,等.左归丸调控DKK1靶点防治糖皮质激素性骨质疏松[J].中国组织工程研究,2018,22(16):2520-2525.
[37] Saarto T,Blomqvist C,Valimaki M,et al. Chemical castration induced by adjuvant cyclophosphamide,methotrexate, and fluorouracil chemotherapy causes rapid bone loss that is reduced by clodronate:a randomized study in premenopausal breast cancer patients[J]. J Clin Oncol,1997,15(4):1341-1347.
[38] Murphy C, Muscat A, Ashley D, et al. Tailored NEOadjuvant epirubicin, cyclophosphamide and nanoparticle albumin-bound paclitaxel for breast cancer:The phaseⅡNEONAB trial-clinical outcomes and molecular determinants of response[J]. PLo S One,2019,14(2):e0210891.
[39] Sugita J,Kagaya Y,Miyamoto T,et al. Myeloablative and reduced-intensity conditioning in HLAhaploidentical peripheral blood stem cell transplantation using post-transplant cyclophosphamide[J]. Bone Marrow Transplant,2019,54(3):432-441.
[40] PEI Y,ZHENG K,SHANG G,et al. Therapeutic effect of strontium ranelate on bone in chemotherapy-induced osteopenic rats via increased bone volume and reduced bone loss[J]. Biol Trace Elem Res,2019,187(2):472-481.
[41] ZHAO D, WANG C, ZHAO Y, et al.Cyclophosphamide causes osteoporosis in C57BL/6 male mice:suppressive effects of cyclophosphamide on osteoblastogenesis and osteoclastogenesis[J].Oncotarget,2017,8(58):98163-98183.
[42]刘政.环磷酰胺致骨质疏松大鼠模型的建立及左归丸的防治作用[D].广州:暨南大学,2015.
[43]彭慧娟,朱玲,许丽绵,等.左归丸对环磷酰胺卵巢早衰大鼠卵巢Bax、Bcl-2及Cyt-c的影响[C]//中国中西医结合学会妇产科专业委员会.第9届中国中西医结合学会妇产科专业委员会第二次学术会议论文集.昆明,2017:8-10.
[44]周家俊,关鑫,赵东.部分肾切除加高磷饮食所致大鼠肾性骨病模型研究[J].中国中西医结合肾病杂志,2002(10):567-569.
[45]袁军,王小琴,马晓红,等.左归丸对5/6肾大部切除模型并肾性骨病大鼠甲状旁腺的影响[J].中医药学报,2013,41(1):76-79.
[46]王小琴,袁军,马晓红,等.左归丸对5/6肾大部切除模型并肾性骨病大鼠骨代谢指标的影响[J].中国中西医结合肾病杂志,2012,13(7):584-586.
[47] LONG F. Building strong bones:molecular regulation of the osteoblast lineage[J]. Nat Rev Mol Cell Biol,2011,13(1):27-38.
[48] Gomes A M,Douglas-de-Oliveira D W,Ferreira S D,et al. Periodontal disease,peri-implant disease and levels of salivary biomarkers IL-1β, IL-10, RANK, OPG,MMP-2,TGF-βand TNF-α:follow-up over 5 years[J].J Appl Oral Sci,2019,27:e20180316.
[49] LI B,Boast S,Kenia de los S,et al. Mice deficient in Abl are osteoporotic and have defects in osteoblast maturation[J]. Nat Genet,2000,24(3):304-308.
[50] Burshell A L,Mricke R,Correa-Rotter R,et al.Correlations between biochemical markers of bone turnover and bone density responses in patients with glucocorticoid-induced osteoporosis treated with teriparatide or alendronate[J]. Bone,2010,46(4):935-939.
[51] Glover S J,Eastell R,McCloskey E V,et al. Rapid and robust response of biochemical markers of bone formation to teriparatide therapy[J]. Bone,2009,45(6):1053-1058.
[52] Akbari Moghaddam Kakhki R,LU Z,Thanabalan A,et al. Eimeria challenge adversely affected long bone attributes linked to increased resorption in 14-day-old broiler chickens[J]. Poult Sci, 2019,98(4):1615-1621.
[53] Kim S K,Lee M H,Rhee M H. Studies on the effects of biomedicinal agents on serum concentration of Ca2+,P and ALP activity in osteoporosis-induced rats[J]. J Vet Sci,2003,4(2):151-154.
[54] YAO H H,TANG S M,WANG Z M,et al. Study of bone mineral density and serum bone turnover markers in newly diagnosed systemic lupus erythematosus patients[J]. J Peking Univ Health Sci,2018,50(6):998-1003.
[55] Hatate K,Kayano M,Kawashima C,et al. The effect of colostrum intake on osteoprotegerin and bone metabolic markers in the blood of newborn calves during the first week of life[J]. J Vet Med Sci,2019,81(1):15-21.
[56] PAN C,LIU X,LI T,et al. Kinetic of bone turnover markers after osteoporotic vertebral compression fractures in postmenopausal female[J]. J Orthop Surg Res,2018,13(1):314.
[57] Rief H,Omlor G,Akbar M,et al. Biochemical markers of bone turnover in patients with spinal metastases after resistance training under radiotherapy-a randomized trial[J]. BMC Cancer,2016,16:231.
[58]叶素敏,谭峰,刘飞祥,等.左归丸对去卵巢大鼠下丘脑Orexin及其受体mRNA表达的影响[J].中国实验方剂学杂志,2019, doi:org/10. 13422/j. cnki.syfjx. 20191021.
[59]童伟伟.基于补肾益癸理论探讨左归丸防治PMOP的疗效机制[D].南京:南京中医药大学,2018.
[60]卢建华,王维佳,储小兵,等.补肾方对去卵巢大鼠Ⅰ型胶原代谢的影响[J].中华中医药杂志,2010,25(7):1109-1111.
[61]孙月娇,任艳玲,曲宁宁,等.左、右归丸及其拆方对去卵巢骨质疏松模型大鼠的比较研究[J].中国骨质疏松杂志,2015,21(8):910-913,921.
[62]李泽钊,邓建强.地仲强骨胶囊治疗肾虚兼气血不足型骨质疏松症的疗效及对骨代谢、骨转化的影响[J].中国实验方剂学杂志,2018,24(8):159-164.
[63] Kanis J A,Cooper C,Rizzoli R,et al. Review of the guideline of the American College of Physicians on the treatment of osteoporosis[J]. Osteoporos Int,2018,29(7):1505-1510.
[64] Hanson N A, Bagi C M. Alternative approach to assessment of bone quality using micro-computed tomography[J]. Bone,2004,35(1):326-333.
[65]刘晓,于文涛,靳贺超,等.补肾通络方对肺纤维化大鼠肺组织TGF-β1/Smads通路的影响[J].中国实验方剂学杂志,2019,25(2):96-102.
[66]卢建华,王维佳,陈明涛,等.补肾方对去卵巢大鼠腰椎骨形态计量学的影响[J].中医正骨,2011,23(5):24-26.
[67]王如然,冯艳华,张玉杰,等.左归丸、右归丸对去卵巢骨质疏松模型大鼠的效用[J].基因组学与应用生物学,2018,37(3):1104-1109.
[68] Massie A M,Kapatkin A S,Garcia T C,et al. Effects of hole diameter on torsional mechanical properties of the rabbit femur[J]. Vet Comp Orthop Traumatol,2019,32(1):51-58.
[69]董苗淼.运动配合左归丸对去卵巢大鼠骨生物力学及相关激素的影响[D].上海:上海体育学院,2010.