来源：风湿病与关节炎,2022,11（8）:1-5,12.

苦参碱对肿瘤坏死因子-α诱导胶原诱导性关节炎大鼠成纤维样滑膜细胞PI3K/AKT信号传导通路的影响（基础研究）

刘　琼¹，丁超华²，白菊芳²，关金花²，孙娜娜²，吕　杨²，张　乐²，王凤春²，杨永生²

　　【摘　要】目的：探讨苦参碱对肿瘤坏死因子-α（TNF-α）诱导胶原诱导性关节炎（CIA）大鼠成纤维样滑膜细胞（FLS）中磷脂酰肌醇-3-激酶（PI3K）/蛋白激酶B（PKB/AKT）信号传导通路相关蛋白及基因表达的影响。方法：按照随机数字表法将40只SD大鼠分为正常对照组、炎症细胞组、苦参碱组、抑制剂组，每组10只。除正常对照组外，其余各组通过牛Ⅱ型胶原与不完全弗氏佐剂建立CIA大鼠模型。断椎法处死大鼠，剥离大鼠滑膜组织，提取FLS，并进行细胞培养、鉴定。正常对照组不加任何药物干预，炎症细胞组仅予TNF-α干预，苦参碱组予TNF-α及苦参碱干预，抑制剂组予TNF-α及LY294002干预。Western Blot测定各组CIA大鼠FLS中p-PI3K、p-AKT、AKT蛋白的表达程度，RT-PCR测定PI3K mRNA、AKT mRNA的表达程度。结果：与正常对照组比较，其余各组CIA大鼠FLS数量明显增多；免疫荧光测定显示绝大多数细胞染色呈阳性表达。与炎症细胞组比较，苦参碱组、抑制剂组p-PI3K、p-AKT蛋白表达程度下降（P < 0.01或P < 0.05），PI3K mRNA、AKT mRNA表达程度受到明显抑制（P < 0.01）；各组间非活化AKT蛋白表达差异无统计学意义（P > 0.05）。结论：苦参碱对CIA大鼠FLS中PI3K/AKT信号传导通路的过度活化具有抑制作用。

　　【关键词】　类风湿关节炎；苦参碱；PI3K/AKT信号传导通路；成纤维样滑膜细胞；胶原诱导性关节炎

Effect of Matrine on PI3K/AKT Signaling Pathway of Fibroblast Like Synovial Cells of TNF-α Induced CIA Rats

LIU Qiong,DING Chao-hua,BAI Ju-fang,GUAN Jin-hua,SUN Na-na,LYU Yang,ZHANG Le,WANG Feng-chun,YANG Yong-sheng

　　【ABSTRACT】 Objective:To investigate the effect of matrine on PI3K/AKT signaling pathway of fibroblast like synovial cells of TNF-α induced CIA rats.Methods:According to the random number table,40 SD rats were divided into a normal control group,an inflammatory cell group,a matrine group and an inhibitor group,with 10 rats in each group.Except the normal control group,the CIA rat models were established in the other groups by bovine typeⅡcollagen and incomplete Freund's adjuvant.The rats were killed by breaking push method,and the synovial tissue of the rats was stripped.FLS was extracted,and the cells were cultured and identified.The normal control group did not receive any drug intervention,the inflammatory cell group only received TNF-α intervention,the matrine group was given TNF-α and matrine intervention,and the inhibitor group was given TNF-α and LY294002 intervention.The expressions of p-PI3K,p-AKT and AKT proteins in FLS of CIA rats in each group were determined by Western Blot.The expressions of PI3K mRNA and AKT mRNA were determined by RT-PCR.Results:Compared with the normal control group,the number of FLS of CIA rats in other groups increased significantly.Immunofluorescence assay showed that most of the cells were positive.Compared with the inflammatory cell group,the expressions of p-PI3K and p-AKT protein in the matrine group and the inhibitor group decreased（P < 0.01 or P < 0.05）,and the expressions of PI3K mRNA and AKT mRNA were significantly inhibited（P < 0.01）.There was no significant difference in the expression of non activated AKT protein among the groups（P > 0.05）.Conclusion:Matrine can inhibit the over activation of PI3K/AKT signaling pathway in FLS of CIA rats.

　　【Keywords】 rheumatoid arthritis;matrine;PI3K/AKT signaling pathway;fibroblast like synovial cells;collagen induced arthritis