针刀对膝骨性关节炎兔软骨细胞外基质Ⅱ型胶原、聚集蛋白聚糖相关蛋白表达的影响  

Effects of Acupotomy Intervention on Regional Pathological Changes and Expression of Cartilage-mechanics Related Proteins in Rabbits with Knee Osteoarthritis

作　　者：梁楚西 郭妍[1] 陶琳[2] 肖红[3] 刘清国[1] 马慧芳[1] 郭长青[1] 

机构地区：[1]北京中医药大学针灸推拿学院,北京100029 [2]北京中医药大学东方医院,北京100078 [3]石家庄市第一医院,石家庄050011

出　　处：《针刺研究》2015年第2期119-124,140,共7页Acupuncture Research

基　　金：国家自然科学基金资助项目(No.81173333);北京中医药大学自主选题项目(No.2013-JYBZZ-XS-042)

摘　　要：目的:观察针刀对膝骨关节炎(knee osteoarthritis,KOA)兔软骨细胞外基质Ⅱ型胶原(Col-Ⅱ)、聚集蛋白聚糖(Aggrecan)相关蛋白表达的影响,探讨针刀治疗KOA的相关机制。方法:新西兰兔随机分为空白组、模型组、针刀组和电针组,每组10只,以左后肢伸直位固定制动法制备KOA模型。针刀组以膝关节内、外侧副韧带及髌韧带为松解对象,以纵疏横剥法进行针刀松解,每周1次,共治疗3周;电针组电针左后肢"阴陵泉""阳陵泉""内膝眼""外膝眼",每次20min,每周治疗3次,共3周。X线观察膝关节影像学改变,光镜观察关节软骨形态学改变,Western blot法分别检测膝关节整合素β1(Integrinβ1)、Col-Ⅱ、基质金属蛋白酶3(MMP-3)及Aggrecan的蛋白表达水平。结果:X线结果显示,造模后,模型制备成功,关节病损情况属于早中期膝骨关节炎。光镜结果显示,模型组关节软骨层表面粗糙不平,表层软骨细胞稀少,细胞排列紊乱,部分软骨细胞坏死,潮线模糊或扭曲不完整,部分软骨有血管通过或血管翳生成,电针组及针刀组关节软骨组织结构均较模型组明显改善,针刀组较电针组改善更明显。Mankin评分比较,模型组较空白组明显升高(P<0.01),针刀组较模型组明显降低(P<0.01)。Western blot结果显示,模型组膝关节组织中Integrinβ1、Col-Ⅱ、Aggrecan蛋白表达较空白组显著降低(P<0.01),MMP-3蛋白表达显著升高(P<0.01);针刀组Integrinβ1、Col-Ⅱ、Aggrecan蛋白表达较模型组显著升高(P<0.01,P<0.05),MMP-3蛋白表达较模型组显著降低(P<0.01);电针组Integrinβ1蛋白表达较模型组明显升高(P<0.05),其它指标与模型组的差异均无统计学意义(P>0.05)。结论:针刀可能通过对软骨力学环境的影响,激活整合素力学信号转导通路,促进细胞外基质Col-Ⅱ、Aggrecan蛋白表达,下调MMP-3的蛋白表达水平,阻抑软骨细胞外基质的降解,延缓软骨损伤与关节退变程度,达到治疗KOA的�Objective To observe the effect of acupotomy （needle-knife） therapy on local pathological changes and cartilage-mechanics related protein expression in rabbits with knee osteoarthritis （KOA） so as to study its mechanisms underlying improving KOA. Methods Forty New Zealand rabbits were randomly divided into normal control group, model group, acupoto- my group, and electroacupuncture （EA） group （n = 10 in each group）. The KOA model was established by immobilization of the left knee-joint （modified Videman method） for 6 weeks. After modeling, acupotomy relaxing was applied to the lateral collateral ligament and patellar ligament of the left knee-joint, once a week for 3 times, and EA （2 Hz/100 Hz, 3 mA） was applied to the left ＂Yanglingquan＂ （GB 34）, ＂Yinlingquan＂ （SP 9）, ＂Neixiyan＂ （EX-LE 4） and ＂Waixiyan, （ST 35） for 20 min, 3 times a week for three weeks. The expression levels of Integrin 15 1, type II collagen （Col-Ⅱ ） , matrix metalloproteinase （MMP）-3 and Aggrecan proteins of the cartilage tissue of the left femoral medial and external condyles were observed by Western blot. Pathological chan- ges of the knee-joint by X-ray scanning and those of the femoral condyle tissue were evaluated by Mankin＇s scores under light microscope after H.E. staining. Results X-ray showed successful modeling, and pathological changes of the articular cartilage belonged to the early and moderate lesion of knee osteoarthritis. ＂The Mankin＇s score was significantly higher in the model group than in the control group （P〈0. 01）, and after the treatment, the Mankin＇s scores were significantly decreased in the aoupotomy group （P〈0.01）, rather than in the EA group （P〉0.05）. The results of Western blot showed that after modeling, the expression levels of Integrin β1, Col-Ⅱ and Aggrecan proteins of the femoral articular cartilage were considerably decreased （P〈0. 01 ）, while that of MMP-3 protein was significantly increased （P〈0. 01 ）.

关 键 词：针刀 膝骨性关节炎 病理变化 关节软骨 整合素Β1 基质金属蛋白酶-3 Ⅱ型胶原聚集蛋白聚糖 

分 类 号：R244.9[医药卫生—针灸推拿学]