补肾活血中药“杜仲-当归”治疗骨性关节炎的作用及机制研究  

Study onEffect and Preparation of Eucommia ulmoides and Angelica sinensisfor Tonifying Kidney and Activating Blood Circulation in Treatment of Osteoarthritis

作　　者：孙亮亮 章煌杰 鲁琛[3] 应建伟 叶承锋 SUN Liangliang;ZHANG Huangjie;LU Chen;YING Jianwei;YE Chengfeng(Zhejiang Hospital,Hangzhou 310013,Zhejiang,China;Zhejiang Cancer Hospital,Hangzhou 310022,Zhejiang,China;The Second Affiliated Hospital of Zhejiang University,Hangzhou 310009,Zhejiang,China;Tonglu County Hospital of Traditional Chinese Medicine,Hangzhou 311500,Zhejiang,China)

机构地区：[1]浙江医院,浙江杭州310013 [2]浙江省肿瘤医院,浙江杭州310022 [3]浙江大学附属第二医院,浙江杭州310009 [4]桐庐县中医院,浙江杭州311500

出　　处：《中华中医药学刊》2019年第11期2639-2644,I0014-I0016,共9页Chinese Archives of Traditional Chinese Medicine

基　　金：国家自然科学基金项目(81273772);浙江省自然科学基金项目(LY18H170003);浙江省中医药科技计划项目(2015ZA178);浙江省医药卫生科技计划项目(2015KYA105)

摘　　要：目的:骨性关节炎(OA)是临床常见病、多发病,目前缺乏切实有效的治疗方法。研究明确补肾活血中药的作用,探寻补肾活血中药对OA的调控机制。方法:通过建立OA模型,以补肾活血中药为代表,进行对MMP13表达的调控研究,OA模型分别灌胃给予补肾药、活血药和补肾活血药,观察各组中药干预后MMP13的回调及回调程度,并结合组织病理学实验结果,明确补肾活血中药的作用:采用HE染色和SO染色观察OA大鼠膝关节软骨的层次、软骨细胞及软骨下骨的变化;ELISA检测大鼠血清和关节滑膜组织IL-1β﹑TNF-α和MMP13的表达情况;免疫组化观察补肾活血中药对OA大鼠膝关节MMP13表达的影响。结果:HE和SO染色初步可判断病变程度从高到低依次为OA模型组、活血组、补肾组、活血补肾组、阳性药物组;ELISA检测显示:OA模型组与正常对照组比较,血清中炎症因子IL-1β﹑TNF-α、MMP13含量明显上升(P<0.01),与OA模型组比较,各给药组显著降低IL-1β﹑TNF-α含量,且补肾活血组血清中的IL-1β﹑TNF-α、MMP13含量要低于单独用药组;免疫组化结果显示:正常组大鼠膝关节未见MMP13阳性表达。OA鼠具有明显的MMP13阳性染色且阳性染色面积和累积光密度值显著升高(P<0.01)。相对于OA模型组,补肾组、活血组、补肾活血组和阳性药物组大鼠MMP13阳性表达、阳性染色面积和累积光密度值均明显降低,降低程度从高到低依次为阳性药物组、补肾活血组、补肾组、活血组。结论:补肾活血中药(杜仲+当归)对OA具有明确治疗作用,且补肾活血组优于补肾组、活血组,揭示补肾活血治则意义,其治疗主要机制可能是通过降低关节软骨中MMP13含量以及血清中的IL-1β和TNF-α含量。Objective:Osteoarthritis(OA)is a common and frequently-occurring disease,and there is no effective treatment.The aim of this study is to clarify the role of Chinese herbs for invigorating kidney and activating blood circulationandexplore the regulatory mechanismon OA.Methods:By establishing the OA model and taking the Chinese herbal medicine for invigorating kidney and activating blood as the representative,the regulation of the expression of MMP13 was studied.The OA model was administered the tonifying kidneymedicine,the blood activating medicine and the tonifying kidney and activating blood medicine respectively,and observed the callback and the degree of the callus of the Chinese medicine after the intervention of the traditional Chinese medicine,and combined with the histopathological results of the histopathology to make clear the Chinese herbal medicine for invigorating the kidney and activating blood.HE staining and SO staining were used to observe the levels of cartilage in the knee joint,cartilage cells and subchondral bone in OA rats.ELISA was used to detect the expressions of IL-1β,TNF-αand MMP13 in the sera and synovial tissues of rats,and the effect of tonifying kidney and activating blood medicine on the MMP13 expression of knee joint in OA rats was observed by immunohistochemistry.Results:HE and SO staining showed that the degree of pathological changes from high to low was in the order of the model group,the Huoxue group,the kidney-tonifying group,the activating blood group and the positive drug group.The ELISA test showed that the serum levels of IL-1β,TNF-αand MMP13 in the serum were significantly increased(P<0.01)in the OA model group and the normal group(P<0.01).The contents of IL-1βand TNF-αwere significantly reduced in the drug group,and the contents of TNF-A and MMP13 in the serum of Eucommia+Angelica group(tonifying kidney and activating blood group)was lower than that of the single drug group.The results of immunohistochemistry showed that the normal group had no positive expression

关 键 词：MMP13 OA 补肾活血中药 IL-1Β TNF-α 

分 类 号：R684.3[医药卫生—骨科学]