基于线粒体凋亡途径探究经筋刺法对膝骨关节炎模型大鼠股四头肌保护作用机制  

Study on the Protective Mechanism of Meridian and Tendon Acupuncture on the Quadriceps Femoris Cells in Rat Model of Knee Osteoarthritis Based on Mitochondrial Apoptosis Pathway

作　　者：郑曲 董宝强[1,3] 林星星 张宇[1] 关雪峰 韩易言[1,3] ZHENG Qu;DONG Baoqiang;LIN Xingxing;ZHANG Yu;GUAN Xuefeng;HAN Yiyan(Liaoning University of Traditional Chinese Medicine,Shenyang 110847,China;Key Laboratory of Acupuncture and Moxibustion Biology,Department of Education of Liaoning Province,Shenyang 110847,China;Key Laboratory of Acupuncture and Moxibustion Health and Rehabilitation of Liaoning Province,Shenyang 110847,China;Shenyang Pharmaceutical University,Shenyang 110016,China)

机构地区：[1]辽宁中医药大学,辽宁沈阳110847 [2]辽宁省教育厅针灸生物学重点实验室,辽宁沈阳110847 [3]辽宁省针灸养生康复重点实验室,辽宁沈阳110847 [4]沈阳药科大学,辽宁沈阳110016

出　　处：《中国中医药信息杂志》2025年第4期92-99,共8页Chinese Journal of Information on Traditional Chinese Medicine

基　　金：国家重点研发计划(2021YFC1712800);辽宁省科技厅创新能力提升联合基金(2022-NLTS-13-02;2022-NLTS-13-04);国家资助博士后研究人员计划(GZC20231025);辽宁省教育厅基本科研项目(JYTQN2023458)。

摘　　要：目的观察经筋刺法对膝骨关节炎(KOA)模型大鼠股四头肌细胞线粒体凋亡的影响,探究其治疗KOA的作用机制。方法将40只SPF级健康雄性Wistar大鼠随机分为假手术组(10只)和造模组(30只),造模组采用改良Hulth方法建立KOA大鼠模型。将成模大鼠随机分为模型组、塞来昔布组和经筋刺法组,每组9只,分别予相应措施干预,连续14 d。改良Lequesne MG评分评价大鼠膝关节功能,测量大鼠患肢大腿围度、股四头肌湿重、湿重维持率及湿重比,HE染色观察股四头肌组织形态,TUNEL法检测股四头肌细胞凋亡情况,免疫荧光法检测股四头肌组织活性氧(ROS)、超氧化物歧化酶(SOD)、胱天蛋白酶(Caspase)-3、Caspase-9表达,荧光双染法检测股四头肌组织ROS与Caspase-3共表达情况,Western blot检测股四头肌组织凋亡相关蛋白表达。结果与假手术组比较,模型组大鼠膝关节改良Lequesne MG评分升高(P<0.05),患肢大腿围度、股四头肌湿重、湿重维持率及湿重比降低(P<0.05),股四头肌肌纤维排列紊乱、松散,部分肌纤维溶解、坏死,伴大量炎性渗出,淋巴细胞增多,细胞凋亡指数升高(P<0.05),股四头肌组织ROS、Caspase-3、Caspase-9表达升高,SOD表达降低(P<0.05),Bcl-2、Bcl-XL、Bax、细胞色素C(CytC)、Caspase-3、Caspase-9蛋白表达升高(P<0.05);与模型组比较,塞来昔布组和经筋刺法组大鼠膝关节改良Lequesne MG评分降低(P<0.05),患肢大腿围度、股四头肌湿重、湿重维持率及湿重比升高(P<0.05),股四头肌肌纤维结构正常,肌膜较完整,细胞凋亡指数降低(P<0.05),股四头肌组织ROS、Caspase-3、Caspase-9表达降低,SOD表达升高(P<0.05),股四头肌组织Bcl-2、Bcl-XL、Bax、CytC、Caspase-3、Caspase-9蛋白表达降低(P<0.05)。荧光双染ROS与Caspase-3表达趋势一致。结论经筋刺法能通过降低KOA模型大鼠股四头肌组织ROS表达抑制线粒体凋亡相关蛋白表达,进而改善骨骼肌肌力,发挥一定�Objective To investigate the effects of meridian and tendon acupuncture on the mitochondrial apoptosis of quadriceps femoris muscle cells in model rats of knee osteoarthritis(KOA);To explore its mechanism for the treatment of KOA.Methods Totally 40 SPF healthy male Wistar rats were randomly divided into sham-operation group(10 rats)and modeling group(30 rats).The modified Hulth method was used to establish a KOA rat model.The model rats were randomly divided into model group,celecoxib group and meridian and tendon acupuncture group,with 9 rats in each group.Corresponding intervention measures were given to each group for 14 consecutive days.Improved Lequesne MG score was used for evaluating knee joint function in rats,measuring thigh circumference,quadriceps femoris wet weight,wet weight maintenance rate and wet weight ratio of the affected limb,HE staining was used to observe the morphology of quadriceps femoris tissue,TUNEL method was used to detect apoptosis of quadriceps femoris cells,immunofluorescence was used to detect the expressions of reactive oxygen species(ROS),superoxide dismutase(SOD),Caspase-3 and Caspase-9 in quadriceps femoris muscle tissue,fluorescence double staining method was used to detect the co-expression of ROS and Caspase-3 in quadriceps femoris tissue,Western blot was used to detect the expressions of apoptosis related proteins in quadriceps femoris tissue.Results Compared with the sham-operation group,the improved Lequesne MG score of the knee joint in the model group rats increased(P<0.05),the circumference of the thigh,wet weight of the quadriceps femoris,wet weight maintenance rate and wet weight ratio of the affected limb decreased(P<0.05),the arrangement of quadriceps femoris muscle fibers was disordered and loose,with some muscle fibers dissolved and necrotic,accompanied by a large amount of inflammatory exudate,an increase in lymphocytes,and an increase in cell apoptosis index(P<0.05),the expressions of ROS,Caspase-3 and Caspase-9 in quadriceps femoris tissue increased,while the

关 键 词：经筋刺法 膝骨关节炎 活性氧 线粒体凋亡途径 大鼠 

分 类 号：R245.3[医药卫生—针灸推拿学]