基于细胞焦亡探讨中药复方对CIA大鼠炎症的影响  

Study on Effect of Chinese Medicine Compound on Inflammation of CIA Rats Based on Cell Charring

作　　者：尚碧月 姜泉[1] 夏聪敏 翟争 王建[1] 张富渊 姚传辉 巩勋[1] 唐晓颇[1] SHANG Biyue;JIANG Quan;XIA Congmin;ZHAI Zheng;WANG Jian;ZHANG Fuyuan;YAO Chuanhui;GONG Xun;TANG Xiaopo(Guang’anmen Hospital,China Academy of Chinese Medical Sciences,Beijing 100053,China;Shanghai Taikuntang Traditional Chinese Medicine Hospital,Shanghai 200000,China)

机构地区：[1]中国中医科学院广安门医院,北京100053 [2]上海泰坤堂中医医院,上海200000

出　　处：《中国中医基础医学杂志》2023年第4期593-597,共5页JOURNAL OF BASIC CHINESE MEDICINE

基　　金：国家自然科学基金项目(81974568)-基于成破骨细胞平衡探讨清热活血方药调控miRNA-214-ATF4通路抑制类风湿关节炎骨破坏的机制研究;国家自然科学基金项目(82230121)-中西医结合防治类风湿关节炎骨破坏的免疫机制研究;国家重点研发计划项目(2018YFC1705202)-中医药治疗类风湿关节炎的临床队列及疗效评价研究;中国中医科学院科技创新工程项目(CI2021A01501)-活动性类风湿关节炎核心证候的中医药方案优化及优势人群特征研究。

摘　　要：目的 探讨清热活血方基于NOD样受体蛋白3(nod-like receptor protein 3,NLRP3)/半胱氨酸天冬氨酸蛋白水解酶-1(cystein-containing aspartat-specific protease-1,Caspase-1)/消皮素D(gasdermin D,GSDMD)细胞焦亡通路治疗类风湿关节炎(rheumatoid arthritis,RA)大鼠炎症的作用机制。方法 60只sprague-dawley(SD)大鼠随机分为正常组,模型组,清热活血方低(8.5 g/kg)、中(17 g/kg)、高剂量组(34 g/kg),甲氨蝶呤组(0.2 mg/kg),除正常组外,各组大鼠尾根部皮下注射牛Ⅱ型胶原乳剂构建胶原诱导性关节炎大鼠(collagenⅡ-induced arthritis,CIA)模型,造模成功后予对应药物灌胃(正常组、模型组予0.9%氯化钠溶液灌胃),连续灌胃30 d后观察不同实验组大鼠体质量及足趾容积变化,ELISA法检测血清白细胞介素(interleukin,IL)-1β、IL-18水平,用荧光定量PCR检测各组大鼠膝关节滑膜组织NLRP3/Caspase-1/GSDMD mRNA表达水平。结果 与正常组比较,模型组大鼠血清IL-1β、IL-18水平及滑膜组织NLRP3/Caspase-1/GSDMD mRNA表达水平均显著升高(P<0.01);与模型组比较,各药物干预组血清IL-1β、IL-18水平均明显下降(P<0.01),滑膜组织NLRP3/Caspase-1/GSDMD mRNA表达水平显著降低(P<0.01,P<0.05)。结论 清热活血方可能通过下调细胞焦亡通路NLRP3/Caspase-1/GSDMD mRNA表达及减少IL-1β、IL-18炎症因子释放,改善炎症状态,减轻CIA大鼠炎症,减缓RA进展。Objective To investigate the mechanism of action of Qingre Huoxue Formula based on nod-like receptor protein 3(NLRP3)/cystein-containing aspartat-specific protease-1(Caspase-1)/gasdermin D(GSDMD)cell pyroptosis pathway in treating inflammatory response in rheumatoid arthritis rats.Methods Sixty sprague-dawley(SD)rats were randomly divided into blank group,model group,low(8.5 g/kg),medium(17 g/kg),and high-dose group(34 g/kg)of Qingre Huoxue Formula,methotrexate group(0.2 mg/kg).In addition to the normal group,the collagenⅡ-induced arthritis(CIA)rat model was constructed by subcutaneous injection of bovine type II collagen emulsion at the tail root of each group.After successful modeling,the corresponding drug was given gavage(0.9%sodium chloride solution gavage in normal group and model group),and the changes in body weight and toe volume of rats in different experimental groups were observed after 30 d of continuous gavage,the serum interleukin(IL)-1βand IL-18 levels were detected by ELISA,and the expression levels of NLRP3/Caspase-1/GSDMD mRNA in Synovial tissue of the knee joint of rats were detected by real-time polymerase chain reaction.Results Compared with the normal group,the serum IL-1β,IL-18 levels and NLRP3/Caspase-1/GSDMD mRNA expression levels of synovial tissues in the model group were significantly increased(P<0.01).Compared with the model group,the serum levels of IL-1βand IL-18 in each drug intervention group decreased significantly(P<0.01).NLRP3/Caspase-1/GSDMD mRNA expression levels in rat synovial tissue were significantly reduced(P<0.01,P<0.05).Conclusion Qingre Huoxue Formula can improve the inflammatory state,reduce inflammation in CIA rats,and delay the progression of rheumatoid arthritis by reducing the activity of NLRP3 inflammasomes,inhibiting the expression of NLRP3/Caspase-1/GSDMD mRNA in the cell pyroptosis pathway and reduce the release of IL-1βand IL-18 inflammatory factors.

关 键 词：类风湿关节炎 清热活血方 中药复方 细胞焦亡 NLRP3 CASPASE-1 GSDMD 炎症 

分 类 号：R285.5[医药卫生—中药学]