从肝论治取穴针灸治疗膝骨关节炎模型大鼠关节功能与昼夜节律的关系

Relationship between joint function and circadian rhythm in a rat model of knee osteoarthritis undergoing acupuncture and moxibustion at acupoints selected based on liver theory

背景:膝骨关节炎发病机制复杂,近年来有研究发现昼夜节律与膝骨关节炎关系密切,而肝与昼夜节律有一定联系,从肝论治取穴针灸影响昼夜节律的研究鲜见。目的:探讨从肝论治针灸治疗膝骨关节炎对关节功能的影响及其与昼夜节律的关系。方法:随机将40只3月龄SPF级雄性SD大鼠分为正常组、模型组、常规取穴组和从肝论治取穴组,每组10只。除正常组外,其余3组均制备右膝关节炎模型造模;常规取穴组取足三里+内、外膝眼;从肝论治取穴组取阳陵泉+肝俞+太冲。从造模术后第1周开始,每周进行旷场实验、足底痛阈测试实验和右膝关节活动度测量;治疗结束后,苏木精-伊红染色观察软骨组织病理结构,RT-PCR与免疫荧光法检测软骨组织中Bmal1、Clock基因与蛋白的表达。结果与结论:①与正常组相比,模型组大鼠平均速度、总运动距离明显降低,休息时间明显延长(P <0.05),足底痛阈明显降低(P <0.05),关节活动度下降(P <0.05),组织学评分明显增高(P <0.05),且软骨组织中Bmal1、Clock基因与蛋白的表达均明显下降(P <0.01);②与模型组相比,常规取穴组和从肝论治取穴组各项指标均有改善(P <0.05),且从肝论治取穴组各项指标改善更明显(P <0.05);③提示从肝论治取穴可能通过影响软骨细胞昼夜节律,修复软骨组织,减轻膝骨关节炎疼痛,恢复关节功能。

BACKGROUND:The pathogenesis of knee osteoarthritis is complex.In recent years,studies have found that circadian rhythm is closely related to knee osteoarthritis,and the liver is related to circadian rhythm to some extent.However,there are rare reports on the influence of acupuncture and moxibustion on the circadian rhythm from the liver theory.OBJECTIVE:To explore the relationship between circadian rhythm and joint function of the articular cartilage when treating osteoarthritis with acupuncture at acupoints associated with the liver.METHODS:Forty 3-month-old male SPF Sprague-Dawley rats were randomly divided into normal group,model group,conventional acupuncture group and acupuncture at acupoints associated with the liver group,with 10 rats in each group.In the latter three groups,the osteoarthritis model was constructed in the right knee joint of rats by surgical modeling.Zusanli+Neixiyan+Waixiyan were selected in the conventional acupuncture group,Yanglinquan+Ganshu+Taichong were selected in the group of acupuncture at acupoint associated with the liver.Open field test,plantar pain threshold test and range of motion of the right knee joint were performed and detected since the 1 st week from modeling.After treatment,the pathological structure of cartilage tissues was observed by hematoxylin-eosin staining.RT-PCR and immunofluorescence were used to detect the expression of Bmal1 and Clock gene and protein in cartilage tissues.RESULTS AND CONCLUSION:Compared with the normal group,in the model group,the average speed and total movement distance were significantly reduced,the rest time was significantly prolonged(P<0.05),the plantar pain threshold was significantly reduced(P<0.05),the range of motion of the knee joint was reduced(P<0.05),the histological score was significantly increased(P<0.05),and the mRNA and protein expressions of Bmal1 and Clock were significantly reduced(P<0.01).Compared with the model group,all indexes were improved in the conventional acupuncture group and in the group of acupuncture at acupoints associated with the liver(P<0.05),and a better improvement of each index was in the group of acupuncture at acupoints associated with the liver(P<0.05).Therefore,it is possible to repair cartilage tissue,relieve osteoarthritic pain and restore joint function by influencing circadian rhythm in chondrocytes.