来源：风湿病与关节炎,2020,9（10）:1-5,11.

麝香乌龙丸对人脐静脉内皮细胞的细胞连接及RhoA/ROCK信号通路的影响（基础研究）

梅晓峰¹，王志文²，马炜秀¹，张凯淇¹，侯淋飞¹，曹　颖¹

　　【摘　要】目的：观察麝香乌龙丸对高浓度1-磷酸鞘胺醇（S1P）诱导下人脐静脉内皮细胞（HUVEC）的细胞连接及RhoA/RhoA激酶（ROCK）信号通路的影响，探讨麝香乌龙丸治疗类风湿关节炎的作用机制。方法：将HUVEC分为4组：对照组，高浓度S1P组，高浓度S1P + 麝香乌龙丸组，高浓度S1P + S1PR2拮抗剂JTE013组。采用免疫荧光检测HUVEC中细胞连接相关蛋白ROCK、ZO-1的变化，Western blot检测RhoA、ROCK、磷酸化RhoA激酶（p-ROCK）、ZO-1蛋白的表达水平。结果：高浓度S1P干预后，HUVEC中RhoA、p-ROCK、ROCK蛋白表达均上调，ZO-1表达下调，与对照组比较，差异有统计学意义（P < 0.05）。麝香乌龙丸能够下调RhoA、ROCK、p-ROCK蛋白的表达，上调ZO-1表达，与高浓度S1P组比较，差异有统计学意义（P < 0.05）。结论：麝香乌龙丸能够通过RhoA/ROCK通路调控HUVEC细胞连接相关蛋白ROCK、p-ROCK及ZO-1的表达，稳定HUVEC的血管屏障，这可能是麝香乌龙丸治疗类风湿关节炎的机制之一。

　　【关键词】　关节炎，类风湿；麝香乌龙丸；人脐静脉内皮细胞；S1P细胞连接蛋白；RhoA/ROCK信号通路

Effects of Shexiang Wulong Wan（麝香乌龙丸）on Cell Junction and RhoA/ROCK Signaling Pathway of Human Umbilical Vein Endothelial Cells

MEI Xiao-feng,WANG Zhi-wen,MA Wei-xiu,ZHANG Kai-qi,HOU Lin-fei,CAO Ying

　　【ABSTRACT】Objective:To observe the effect of Shexiang Wulong Wan（麝香乌龙丸,SXWLW）on the cell junction and RhoA/ROCK signal pathway of human umbilical vein endothelial cells（HUVEC）induced by high concentration of sphingosine 1-phosphosphingol（S1P）,and to explore the mechanism of SXWLW in the treatment of rheumatoid arthritis.Methods:HUVEC was divided into four groups:the control group,the high-concentration S1P group（the HS1P group）,the high concentration S1P + SXWLW group（the HS1P + SXWLW group）,and the high concentration S1P + S1PR2 antagonist JTE013 group（the HS1P + JET013 group）.Immunofluorescence was used to detect the changes of cell junction related proteins ROCK and ZO-1 in HUVEC.The expression levels of RhoA,ROCK,phosphorylated RhoA kinase（p-ROCK）and ZO-1 were detected by Western blot.Results:Under the intervention of high concentration of S1P,the expressions of RhoA,p-ROCK and ROCK in HUVEC were up-regulated and ZO-1 was down regulated,the difference was statistically significant compared with the control group（P < 0.05）. SXWLW could down regulate the expression of RhoA,ROCK,p-ROCK protein,and up regulate the expression of ZO-1,and the difference was statistically significant com-pared with the HS1P group（P < 0.05）.Conclusion:SXWLW can regulate the expressions of ROCK,p-ROCK and ZO-1 and stabilize the vascular barrier of HUVEC through RhoA/ROCK pathway,which may be one of the mechanisms of SXWLW in treating rheumatoid arthritis.

　　【Keywords】 arthritis,rheumatoid;Shexiang Wulong Wan（麝香乌龙丸）;HUVEC;S1P cell junction related proteins;RhoA/ROCK signaling pathway