穿石通痹汤通过调节GABRB1-SOCS1/JAK1/STAT3信号途径对CIA模型大鼠关节炎症损伤的影响  

Effects of Mianan Fang on Serum Expression of IL-6,IL-1β,TNF-α and Hippocampal Tissue BDNF Protein in Rats with Depression and Sleep Disorders

作　　者：顾文[1] 张正菊[2] 马卫国[3] 吴子华 白华 张红红 孟凤仙[3] 刘慧[3] GU Wen;ZHANG Zheng-ju;MA Wei-guo;WU Zi-hua;BAI Hua;ZHANG Hong-hong;MENG Feng-xian;LIU Hui(Beijing Hospital of Traditional Chinese Medicine,Beijing 100010;Beijing University of Chinese Medicine,Beijing 100029;Dongfang Hospital,Beijing University of Chinese Medicine,Beijing 100007;Guangwai Hospital,Xicheng District,Beijing 100055;Shunyi Hospital,Capital Medical University,Beijing 101300)

机构地区：[1]首都医科大学附属北京中医医院,北京100010 [2]北京中医药大学,北京100029 [3]北京中医药大学东方医院,北京100078 [4]北京市西城区广外医院,北京100055 [5]北京首都医科大学附属顺义医院,北京101300

出　　处：《世界中西医结合杂志》2024年第5期915-926,共12页World Journal of Integrated Traditional and Western Medicine

基　　金：国家自然科学基金资助项目(51772032);北京中医医院院级课题资助项目(YJ-201808)。

摘　　要：目的 探讨穿石通痹汤对胶原诱导性关节炎(Collagen II induced arthritis,CIA)模型大鼠关节滑膜免疫炎性损伤的疗效及作用机制。方法 SPF级雄性SD大鼠,体质量(200±10)g,采用大鼠尾根部注射牛CⅡ方法制备CIA模型,将成模大鼠按随机数字表法分为模型组、阳性药组、穿石通痹汤组,每组各6只,另取6只未造模大鼠作为正常组。正常组与模型组隔日予去离子水10 ml/kg灌胃,阳性药组予甲氨蝶呤注射液每周0.5 mg/kg腹腔注射,穿石通痹汤组按生药量11.2 g/kg隔日灌胃给药,连续干预4周后处死大鼠,采集血浆、膝关节滑膜及踝关节标本。以HE染色观察大鼠踝关节病理变化;采用Westernblot检测大鼠膝关节滑膜pJAK1、JAK1、p38、GABRB1的蛋白表达水平;ELISA检测血浆IL-6,STAT3,SOCS1表达水平;并采用UHPLC-Q-Exactive Orbitrap MS分析血浆代谢物成分。结果 与正常组比较,模型组大鼠踝关节滑膜明显增生、严重骨质破坏以及炎性细胞浸润;与模型组比较,穿石通痹汤组大鼠关节骨质破坏、关节病变整体评分明显改善。与正常组比较,模型组GABRB1、SOCS1蛋白表达水平显著降低(P<0.01),p38、pJAK1、IL-6,STAT3蛋白表达水平明显升高(P<0.01,P<0.05);与模型组比较,穿石通痹汤组及阳性药组GABRB1、SOCS1蛋白表达水平均显著上调(P<0.01,P<0.05),并明显抑制p38、pJAK1、IL-6,STAT3的蛋白表达(P<0.01),且穿石通痹汤组对GABRB1的上调作用及pJAK1抑制作用显著优于阳性药组(P<0.01);血浆代谢物分析发现甘油磷脂代谢、花生四烯酸代谢、鞘脂代谢等代谢途径及其中50个代谢产物与本病发生密切相关。结论 穿石通痹汤能有效改善CIA模型大鼠关节滑膜免疫炎性代谢性损伤,其机制可能与调节GABRB1-SOCS1/JAK1/STAT3信号途径,调控脂质代谢途径相关。Objective Investigate the efficacy and mechanism of Chuanshi Tongbi decoction on the immune-inflammatory injury of the synovial membrane in collagen II-induced arthritis(CIA)model rats.Methods Specific pathogen-free(SPF)male Sprague-Dawley(SD)rats,weighing 200 ± 10 g,were used to establish the CIA model by injecting bovine CII into the tail root.These model rats were divided into model group,positive drug group,and Chuanshi Tongbi treated decoction group according to the random number table,with 6 rats in each group.Another 6 unmodeled rats were taken as a normal group.The normal group and the model group were given deionized water by gavage at a dose of 10 ml/kg every other day.The positive drug group was given methotrexate at a dosage of 0.5 mg/kg per week by intraperitoneal injection.The Chuanshi Tongbi decoction group was administered crude drug at a dosage of 11.2 g/kg every other day by intragastric.The rats were sacrificed after continuous intervention for 4 weeks,and plasma,knee joint synovial membrane,and ankle joint specimens were collected.The pathological changes in the ankle joints of rats were observed by HE staining.The protein expression levels of pJAK1,JAK1,p38,and GABRB1 in the knee joint synovium of rats were detected by Western blot.The expression levels of IL-6,STAT3,and SOCS1 in plasma were detected by ELISA.UHPLC-Q-Exactive Orbitrap MS was used to analyze the plasma metabolites.Results Compared with the normal group,rats in the model group showed significant synovium proliferation,severe bone destruction,and inflammatory cell infiltration in the ankle joints.Compared with the model group,rats in the Chuanshi Tongbi decoction group exhibited significant improvement in overall scores for joint bone destruction and joint lesions.Compared with the normal group,the expression levels of GABRB1 and SOCS1 protein in the model group were significantly decreased(P<0.01),while the expression levels of p38,pJAK1,IL-6,and STAT3 protein were significantly increased(P<0.01,P<0.05).Compared with the model

关 键 词：穿石通痹汤 胶原诱导性关节炎 GABRB1 SOCS1 免疫炎性损伤 

分 类 号：R285.5[医药卫生—中药学]