温针灸对早期膝骨关节炎软骨下骨中OPG/RANKL水平的影响  

Effects of warm acupuncture on OPG/RANKL level in subchondral bone of early knee osteoarthritis

作　　者：武永利[1,2] 马遇原 刘娣[1] 刘君伟 王铎 WU Yong-li;MA Yu-yuan;LIU Di;LIU Jun-wei;WANG Duo(Department of Orthopedics and Traumatology of TCM,General Hospital of Ningxia Medical University,Yinchuan 750004,China;Key Laboratory of Hui Medicine Modernization of Ministry of Education,Ningxia Medical University,Yinchuan 750004,China;School of TCM,Ningxia Medical University,Yinchuan 750004,China)

机构地区：[1]宁夏医科大学总医院中医骨伤科,银川750004 [2]宁夏医科大学回医药现代化教育部重点实验室,银川750004 [3]宁夏医科大学中医学院,银川750004

出　　处：《中华中医药杂志》2022年第7期4074-4078,共5页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：国家自然科学基金项目(No.81760891);2016年宁夏回族自治区科技创新领军人才项目(No.KJT201610)。

摘　　要：目的:观察温针灸对早期膝骨关节炎(KOA)模型兔软骨下骨中骨保护素(OPG)/核因子κB受体活化因子配体(RANKL)表达水平的影响,探讨温针灸治疗KOA的作用机制。方法:6月龄新西兰雄兔40只,随机分为空白组、模型组、西药组、温针灸组,每组10只。采用管型石膏伸直位固定法建立兔早期KOA模型,经4周实验干预后采用HE染色观察各组软骨及软骨下骨形态结构,免疫组化及Western blot检测各组软骨下骨中OPG、RANKL蛋白的表达,qPCR检测各组软骨下骨中OPG、RANKL mRNA表达。结果:HE染色结果:空白组软骨层表面光滑,软骨细胞排列整齐,软骨及软骨下骨结构完整;模型组软骨层表面粗糙,软骨层明显变薄,骨小梁架构紊乱,软骨细胞排列不均;温针灸组与西药组软骨及软骨下骨病理学表现均有显著改善,软骨细胞较多,分布较为整齐,骨小梁完整。免疫组化与Western blot实验结果:与空白组比较,模型组RANKL蛋白表达显著升高(P<0.01),OPG/RANKL比值显著降低(P<0.01);与模型组比较,温针灸组与西药组OPG蛋白表达显著上调(P<0.01),RANKL蛋白显著降低(P<0.01),OPG/RANKL比值均显著升高(P<0.01)。qPCR实验结果:与空白组比较,模型组RANKL mRNA表达水平显著升高(P<0.01);与模型组比较,温针灸组与西药组OPG mRNA显著上调(P<0.01),RANKL mRNA显著下调(P<0.01),OPG/RANKL比值显著升高(P<0.01)。结论:温针灸治疗早期KOA可通过促进骨保护因子OPG,抑制破骨细胞活化因子RANKL,上调OPG/RANKL比值水平表达,以调节软骨下骨重塑平衡,阻抑软骨下骨退变,延缓KOA发展。Objective:To observe the effects of warm acupuncture on the expression of OPG/RANKL in subchondral bone of rabbit model of early knee osteoarthritis(KOA),and to explore the mechanism of warm acupuncture in treating KOA.Methods:Forty 6-month-old male New Zealand rabbits were randomly divided into control group,model group,western medicine group and warm acupuncture group,with 10 rabbits in each group.Rabbit model of early KOA was established by cast plaster fixation.After 4 weeks of intervention,HE staining was used to observe the morphological structure of cartilage and subchondral bone in each group.The expression of OPG and RANKL protein in subchondral bone was detected by the Immunohistochemistry and Western blot.The mRNA expression of OPG and RANKL in subchondral bone was detected by qPCR.Results:HE staining results:In the blank group,the surface of cartilage layer was smooth,the chondrocytes arranged orderly,and the structure of cartilage and subchondral bone was complete;In the model group,the surface of cartilage layer was rough and the cartilage layer became thinner,the structure of trabecular bone was disordered and the arrangement of chondrocytes was uneven;The pathological features of cartilage and subchondral bone in warm acupuncture group and western medicine group were significantly improved,there were more chondrocytes,more regular distribution and complete trabeculae.The results of immunohistochemistry and Western blot were as follows:compared with the blank group,the expression of RANKL protein in the model group was significantly increased(P<0.01),and the ratio of OPG/RANKL was decreased(P<0.01).Compared with the model group,the expression of OPG protein in warm acupuncture group and western medicine group was significantly increased(P<0.01),RANKL protein was significantly decreased(P<0.01),and OPG/RANKL ratio was significantly increased(P<0.01).The results of qPCR were as follows:compared with the blank group,RANKL mRNA expression level was significantly increased(P<0.01).Compared with the mode

关 键 词：膝骨关节炎 软骨下骨 温针灸 骨保护素 核因子ΚB受体活化因子配体 

分 类 号：R245[医药卫生—针灸推拿学]