电针联合中药浸膏对类风湿性关节炎大鼠疼痛行为、破骨细胞分化及Hedgehog信号通路的影响

Effects of electroacupuncture combined with Chinese herbal extract on pain behavior,osteoclast differentiation and Hedgehog signaling pathway in rats with rheumatoid arthritis

目的:探讨电针联合中药浸膏对类风湿性关节炎大鼠疼痛行为、破骨细胞分化及Hedgehog信号通路的影响。方法:将50只大鼠按照随机数字表法分为正常组、模型组、电针组、中药组及联合组各10只,除正常组外均建立类风湿性关节炎动物模型,建模后,电针组进行针刺治疗,中药组灌胃1mL的12.24g/kg中药浸膏溶液,联合组大鼠进行针刺联合中药浸膏溶液灌胃治疗,其他组别大鼠灌胃等体积的生理盐水,每日1次,连续14d。采用热痛测试仪检测大鼠双侧后足足反射潜伏期(PWTL);运动评分观察各组大鼠运动情况;苏木精-伊红染色观察踝关节病理形态;骨吸收细胞酸性磷酸酶染色(TRAP)染色检测破骨细胞分化数目;免疫印迹技术检测各组大鼠刺猬信号通路(Hh)信号通路相关因子Sonic刺猬信号通路(Shh)、Patched-1(Ptch-1)、脑胶质瘤相关癌基因1(Gli-1)蛋白。结果:末次治疗后,与正常组比较,模型组大鼠PWTL、运动评分降低,破骨细胞分化数目、Shh、Gli1、Ptch1蛋白含量均升高(P<0.05);与模型组比较,电针组及中药组PWTL、运动评分升高,破骨细胞分化数目、Shh、Gli1、Ptch1蛋白含量均降低(P<0.05),电针组及中药组组间比较差异无统计学意义,与电针组及中药组比较,联合组PWTL、运动评分升高,破骨细胞分化数目、Shh、Gli1、Ptch1蛋白含量均降低(P<0.05)。结论:电针联合中药浸膏可显著降低类风湿性关节炎大鼠疼痛感,提高运动能力,同时降低破骨细胞分化程度,研究机制可能与调控Hedgehog信号相关。

Objective To investigate the effects of electroacupuncture combined with traditional Chinese medicine(TCM)decoction on pain behavior,osteoclast differentiation,and Hedgehog signaling pathway in rats with rheumatoid arthritis(RA).Methods:A total of 50 rats were randomly divided into normal group,model group,electroacupuncture group,TCM group,and combination group(n=10 each).The RA animal model was established in all groups except the normal group.After modeling,the electroacupuncture group received needle-stimulation treatment,the TCM group was administered 1 mL of 12.24 g/kg TCM decoction solution by gavage,and the combination group received both treatments.Other groups were gavaged with an equivalent volume of normal saline every day for 14 days.The paw withdrawal thermal latency(PWTL)was measured using a thermal pain tester to evaluate pain behavior.The motor score was observed to evaluate the movement of each group.Ankle joint pathology was observed by H&E staining.Tartrate-resistant acid phosphatase staining was used to detect osteoclast differentiation,and Western blotting was used to detect the expression of Sonic hedgehog(Shh),Patched-1(Ptch-1),and gli-oma-related oncogene homology-1(Gli-1)proteins in the Hedgehog signaling pathway.Results:After the last treatment,compared with the normal group,the PWTL and motor score in the model group were decreased,and the osteoclast differentiation number,and expression of Shh,Gli1,and Ptch1 proteins were increased(P<0.05).Compared with the model group,the PWTL and motor score in the electroacupuncture and TCM groups were increased,while the osteoclast differentiation number,and expression of Shh,Gli1,and Ptch1 proteins were decreased(P<0.05).There was no significant difference between the electroacupuncture group and the TCM group.Compared with the electroacupuncture and TCM groups,the PWTL and motor score in the combination group were increased,while the osteoclast differentiation number,and the expression of Shh,Gli1,and Ptch1 proteins were decreased(P<0.05).Conclusion:Electroacupuncture combined with TCM decoction can significantly reduce pain and enhance movement ability in rats with RA while decreasing osteoclast differentiation.The regulatory mechanism may be related to the Hedgehog signaling pathway.