外膝眼透刺内膝眼温针灸对膝骨性关节炎兔软骨组织Bax、Bcl-2表达的影响
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摘要
目的观察外膝眼透刺内膝眼温针灸对膝骨性关节炎兔软骨组织Bax mRNA、Bcl-2 mRNA及Bax、Bcl-2蛋白表达的影响。方法按随机数字表法将18只新西兰兔随机分为正常对照组(正常组)、膝骨性关节炎组(模型组)和温针内、外膝眼穴组(温针组)。4%木瓜蛋白酶关节腔注射法制作膝骨性关节炎模型。温针组在造模后行内外膝眼穴温针灸治疗20 min,隔天1次,每2周为1个疗程,中间休息2 d,共治疗4个疗程。模型组和正常组不予任何治疗。RT-q PCR法和Western Blotting法检测软骨组织Bax mRNA、Bcl-2 mRNA和Bax、Bcl-2蛋白表达。结果(1)温针灸干预后,新西兰兔的精神、食欲、膝关节活动能力等均明显改善。(2)模型组与正常组比较,膝关节软骨组织Bax mRNA和蛋白表达均显著升高(P<0.001,P<0.01),Bcl-2 mRNA和蛋白表达均明显降低(P<0.001,P<0.001);温针组与模型组比较,温针组Bax mRNA表达显著降低(P<0.05),Bcl-2 mRNA和蛋白表达均明显升高(P<0.05,P<0.05)。结论温针内、外膝眼穴能下调软骨细胞促凋亡基因Bax、上调抗凋亡基因Bcl-2的表达,抑制软骨细胞凋亡通路的信号转导。
Objective To observe the regulatory effects Bax, Bcl-2 mRNA and protein in knee cartilage of rabbits by knee osteoarthritis of warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5). Methods 18 New Zealand rabbits were randomly divided into normal control group( normal group), knee osteoarthritis group( model group) and warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5) group( WNM group) using the randomized controlled Methods. The knee osteoarthritis model was made by injecting four percent of papain into the joint cavity. The WNM group were received warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5) for 20 min, one treatment for two days, every 2 weeks is a course of treatment, there are rest 2 days between 2 course, after 4 courses of treatment. The model group and the normal group were not given any treatment. The expressions of Bax mRNA 、Bcl-2 mRNA and protein in knee cartilage tissue was detected by RT-q PCR and Western-Blotting. Results(1) Compared with the model group, the spirit, appetite and knee joint activities of rabbits in the WNM Group were obviously improved.(2) The expressions of Bax mRNA and protein in the model group were both higher than those in normal group( P<0.001, P<0.01), and the expressions of Bcl-2 mRNA and protein in the model group were both lower than those in normal group( P<0.001, P<0.001). Compared with the model group, the expressions of Bax mRNA in the WNB group were significantly decreased( P <0.05), and the expressions of Bcl-2 mRNA and protein in the model group were both obviously improved( P <0.05, P <0.05). Conclusion Warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5) can down-regulate the expression of Bax mRNA in knee cartilage of rabbits with knee osteoarthritis, and can up-regulate the expression of Bcl-2 mRNA and protein to inhibit the signal transduction of chondrocyte apoptosis pathway.
Objective To observe the regulatory effects Bax, Bcl-2 mRNA and protein in knee cartilage of rabbits by knee osteoarthritis of warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5). Methods 18 New Zealand rabbits were randomly divided into normal control group( normal group), knee osteoarthritis group( model group) and warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5) group( WNM group) using the randomized controlled Methods. The knee osteoarthritis model was made by injecting four percent of papain into the joint cavity. The WNM group were received warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5) for 20 min, one treatment for two days, every 2 weeks is a course of treatment, there are rest 2 days between 2 course, after 4 courses of treatment. The model group and the normal group were not given any treatment. The expressions of Bax mRNA 、Bcl-2 mRNA and protein in knee cartilage tissue was detected by RT-q PCR and Western-Blotting. Results(1) Compared with the model group, the spirit, appetite and knee joint activities of rabbits in the WNM Group were obviously improved.(2) The expressions of Bax mRNA and protein in the model group were both higher than those in normal group( P<0.001, P<0.01), and the expressions of Bcl-2 mRNA and protein in the model group were both lower than those in normal group( P<0.001, P<0.001). Compared with the model group, the expressions of Bax mRNA in the WNB group were significantly decreased( P <0.05), and the expressions of Bcl-2 mRNA and protein in the model group were both obviously improved( P <0.05, P <0.05). Conclusion Warm-needle-moxibustion at neixiyan( EX-LE4) and waixiyan( EX-LE5) can down-regulate the expression of Bax mRNA in knee cartilage of rabbits with knee osteoarthritis, and can up-regulate the expression of Bcl-2 mRNA and protein to inhibit the signal transduction of chondrocyte apoptosis pathway.
引文
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[10]岳萍,高亮,陈默,等.温针灸对膝骨性关节炎兔行为学及关节软骨肿瘤坏死因子-α、基质金属蛋白酶-3含量的影响[J].针刺研究,2016,41(3):235-239.
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[12]叶国平,朱定钰,李俐,等.不同深度温针灸治疗膝骨性关节炎随机对照研究[J].中华中医药杂志,2015,30(8):2886-2889.
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[14]熊勇,彭锐,王华.“双固一通”温针灸实验性膝骨性关节炎模型兔Bcl-2及Bax蛋白的表达[J].中国组织工程研究,2012,16(2):261-264.
[15]刘娣.温针灸对兔膝骨性关节炎软骨细胞JNK信号通路及MMP-1、13表达的影响[D].宁夏:宁夏医科大学,2015.
[16]张翼,吴广文.中医药防治骨性关节炎分子生物学研究进展[J].福建中医药大学学报,2012,22(4):66-69.
[17]林瑞珠,孙改玲,朱宁,等.回医烙灸对兔膝骨关节炎软骨细胞Bcl-2和Bax表达的影响[J].长春中医药大学学报,2017,33(1):34-36.
[18]贾炜.Bcl-2家族研究进展[J].当代医学,2012,18(3):26-28.
[19]朱秀敏.细胞凋亡的形态特征与分子机制研究进展[J].中国老年学杂志,2011,31(13):2595-2597.
[20]OSHIMA Y,AKIYAMA T,HIKITA A,et al.Pivotal role of Bcl-2 family Proteins in the regulation of Chondrocyte apoptosis[J].J Biol Chem,2008,283(39):26499-26508.
[21]WU M X,LI X H,LIN M N,et al.Clinical study on the treatment of knee osteoarthritis of shen-sui insufficiency syndrome type by electroacupuncture[J].Chin J Integr Med,2010,16(4):291-297.
[22]洪昆达,李俐,李西海,等.温针治疗中医不同证型膝骨性关节炎的临床研究[J].中国中西医结合杂志,2012,32(11):1466-1469.
[2]EA H K,MONCEAU V,CAMORS E,et al.Annexin 5overexpression increased articular chondrocyte apoptosis induced by basic calcium phosphate crystals[J].Ann Rheum Dis,2008,67(11):1617-1625.
[3]BRAMONO D S,RICHMOND J C,WEITZEI P P,et al.Matrix metalloproteinases and their clinical applications in orthopaedics[J].Clin Orthop Relat Res,2004,428(428):272-285.
[4]LARSEN C J.The BCL2 gene,prototype of a gene family that controls programmed cell death(apoptosis)[J].Ann Genet,1994,37(3):121-134.
[5]VISSE R,NAGASE H.Matrix metalloproteinases and tissue inhibitors of metalloproteinases structure,function,and biochemistry[J].Circ Res,2003,92(8):827-839.
[6]李忠仁.实验针灸学[M].北京:中国中医药出版社,2003:314.
[7]胡元亮.实用动物针灸手册[M].北京:中国农业出版社,2003:288.
[8]中国国家标准化管理委员会.针灸技术操作规范第20部分:毫针基本刺法[M].北京:中国标准出版社,2009:20-23.
[9]田代华整理.黄帝内经素问[M].北京:人民卫生出版社,2005:104.
[10]岳萍,高亮,陈默,等.温针灸对膝骨性关节炎兔行为学及关节软骨肿瘤坏死因子-α、基质金属蛋白酶-3含量的影响[J].针刺研究,2016,41(3):235-239.
[11]曹锐,杨红玲,何润东.针刺治疗膝骨性关节炎45例临床观察[J].中医杂志,2016,57(13):1133-1136.
[12]叶国平,朱定钰,李俐,等.不同深度温针灸治疗膝骨性关节炎随机对照研究[J].中华中医药杂志,2015,30(8):2886-2889.
[13]孟立强,王卫强,程艳婷.透刺法理论探析[J].江苏中医药,2013,45(1):56-58.
[14]熊勇,彭锐,王华.“双固一通”温针灸实验性膝骨性关节炎模型兔Bcl-2及Bax蛋白的表达[J].中国组织工程研究,2012,16(2):261-264.
[15]刘娣.温针灸对兔膝骨性关节炎软骨细胞JNK信号通路及MMP-1、13表达的影响[D].宁夏:宁夏医科大学,2015.
[16]张翼,吴广文.中医药防治骨性关节炎分子生物学研究进展[J].福建中医药大学学报,2012,22(4):66-69.
[17]林瑞珠,孙改玲,朱宁,等.回医烙灸对兔膝骨关节炎软骨细胞Bcl-2和Bax表达的影响[J].长春中医药大学学报,2017,33(1):34-36.
[18]贾炜.Bcl-2家族研究进展[J].当代医学,2012,18(3):26-28.
[19]朱秀敏.细胞凋亡的形态特征与分子机制研究进展[J].中国老年学杂志,2011,31(13):2595-2597.
[20]OSHIMA Y,AKIYAMA T,HIKITA A,et al.Pivotal role of Bcl-2 family Proteins in the regulation of Chondrocyte apoptosis[J].J Biol Chem,2008,283(39):26499-26508.
[21]WU M X,LI X H,LIN M N,et al.Clinical study on the treatment of knee osteoarthritis of shen-sui insufficiency syndrome type by electroacupuncture[J].Chin J Integr Med,2010,16(4):291-297.
[22]洪昆达,李俐,李西海,等.温针治疗中医不同证型膝骨性关节炎的临床研究[J].中国中西医结合杂志,2012,32(11):1466-1469.