温针疗法联合豨莶草对风湿性关节炎大鼠滑膜细胞活性及SphK1/S1P/S1PR1信号的影响  

Effects of Warm Needling Combined with Siegesbeckiae Herba on Activity of Synovial Cells and SphK1/S1P/S1PR1 Signaling Pathway in Rats with RA

作　　者：文培培 高宇[1] 王晓云[1] 刘益兵[1] 池红万[1] WEN Peipei;GAO Yu;WANG Xiaoyun;LIU Yibing;CHI Hongwan(Zhengzhou Orthopaedic Hospital,Zhengzhou 450052,China)

机构地区：[1]郑州市骨科医院,河南郑州450052

出　　处：《针灸临床杂志》2022年第11期75-81,共7页Journal of Clinical Acupuncture and Moxibustion

基　　金：河南省医学科技攻关计划(联合共建)项目,编号:LHGJ20191139。

摘　　要：目的:探讨温针疗法联合豨莶草对风湿性关节炎大鼠滑膜细胞活性及SphK1/S1P/S1PR1信号的影响。方法:将50只大鼠按照随机数字法分为正常组、RA组、温针组、药物组及联合组,各10只。除正常组外其余大鼠均建立RA大鼠模型,建模成功后,温针组进行温针治疗,药物组大鼠灌胃豨莶草提取物,联合组温针治疗后灌胃豨莶草提取物,其余组灌胃同体积的生理盐水。采用排水法检测各组大鼠足部体积;HE染色观察踝关节病理;提取各组滑膜细胞,MTT法检测存活,Hoechst检测凋亡率;免疫印迹检测SPHK1、S1PR1及ERK1/2蛋白水平。结果:与正常组相比,RA大鼠治疗14 d、18 d及21 d的足部体积均升高(P<0.05),与RA组相比,温针组、药物组及联合组大鼠治疗同一时间足部体积均降低(P<0.05),与温针组比较,联合组较(P<0.05);正常组大鼠踝关节面光滑、滑膜完整、无炎症浸润及水肿,RA组大鼠踝关节炎性细胞浸润严重,滑膜组织增厚,关节腔隙狭窄及软骨破坏,温针组及药物组大鼠踝关节炎性浸润减少,滑膜增生降低,与药物组比较,联合组大鼠踝关节滑膜厚度未见增厚,炎性细胞浸润减少;与正常组比较,RA组滑膜细胞存活OD值升高,凋亡率无意义,与RA组比较,温针组存活OD值降低,凋亡率升高,差异有统计学意义(P<0.05),与温针组比较,联合组滑膜细胞存活OD值降低,凋亡率升高,差异比较有统计学意义(P<0.05);与正常组比较,RA组SPHK1、S1PR1及ERK1/2蛋白升高(P<0.05),与RA组比较,温针组SPHK1、S1PR1及ERK1/2蛋白均降低(P<0.05),与温针组相比,联合组SPHK1、S1PR1及ERK1/2蛋白更低(P<0.05),其余组别差异无统计学意义(P>0.05)。结论:风湿性关节炎大鼠采用温针疗法联合豨莶草干预可抑制滑膜细胞活性,改善大鼠足部体积,这可能与抑制SphK1/S1P/S1PR1信号转导相关。Objective:To investigate the effects of warm needling therapy combined with Siegesbeckiae Herba on the activity of synovial cells and the signal of SphK1/S1P/S1PR1 in rats with rheumatoid arthritis(RA).Methods:50 rats were randomly divided into the normal group,the RA group,the warm needling group,the medication group and the combined group,with 10 rats in each group.RA rat model was established in all other groups except for the normal group.After modeling,the rats in the warm needling group were given warm needling treatment,the rats in the medication group were given Siegesbeckiae Herba extract orally,the rats in the combined group were given Siegesbeckiae Herba extract combined with warm needling,and the rats in the other groups were given the same volume of normal saline orally.Foot volume of rats in each group was measured by drainage method.HE staining was used to observe the pathology of naked joint.Synovial cells were extracted from each group,MTT assay was used to detect cell survival,Hoechst was used to detect apoptosis rate,and the proteins of SPHK1,S1PR1 and ERK1/2 were detected by Western blot.Results:Compared with that in the normal group,the foot volume was increased on 14 d,18 d and 21 d in the RA group(P<0.05);compared with that in the RA group,the foot volume was decreased in the warm needling group,the medication group and the combined group at the same time point(P<0.05);compared with the warm needling group,the above indexes were decreased in the combined group(P<0.05);the naked articular surface was smooth,the synovial membrane was intact,but there was no inflammatory infiltration and edema in the normal group;whereas there was severe infiltration of articular inflammatory cells,thickened synovial tissues,narrow articular spaces and cartilage destruction in the RA group.In the warm needling group and the medication group,the naked joint inflammatory infiltration decreased,and the synovial proliferation decreased.Compared with that in the medication group,the naked joint synovial thickness d

关 键 词：风湿性关节炎 温针 豨莶草 滑膜细胞 SphK1/S1P/S1PR1 

分 类 号：R246.2[医药卫生—针灸推拿学]