红景天苷对TNF-α诱导的人类风湿关节炎成纤维样滑膜细胞增殖的抑制作用及其意义

Inhibitory effect of salidroside on proliferation of HFLS-RA induced by TNF-αand its significance

目的:探讨红景天苷对肿瘤坏死因子α(TNF-α)诱导的人类风湿关节炎成纤维样滑膜细胞(HFLS-RA)增殖的抑制作用,阐明其控制类风湿关节炎(RA)的分子机制。方法:体外培养HFLS-RA,采用TNF-α及不同浓度红景天苷处理细胞,分为正常对照组(0.0μg·L^(-1) TNF-α)、模型对照组(10.0μg·L^(-1)TNF-α)及12.5、25.0、50.0、100.0μmol·L^(-1)红景天苷组(10.0μg·L^(-1) TNF-α+相应浓度红景天苷)。MTT法检测HFLS-RA的增殖活力,ELISA法检测细胞上清液中β-catenin、基质金属蛋白酶7(MMP-7)和Cyclin-D1表达水平,Western blotting法检测细胞中β-catenin蛋白表达水平。结果:与正常对照组比较,模型对照组HFLS-RA增殖活力明显升高(P<0.05)。与模型对照组比较,12.5和25.0μmol·L^(-1)红景天苷组HFLSRA增殖活力虽降低,但组间比较差异无统计学意义(P>0.05);50.0和100.0μmol·L^(-1)红景天苷组HFLSRA增殖活力降低(P<0.05)。与正常对照组比较,模型对照组细胞上清液中β-catenin、MMP-7和Cyclin-D1表达水平明显升高(P<0.05)。与模型对照组比较,12.5和25.0μmol·L^(-1)红景天苷组细胞上清液中β-catenin、MMP-7和Cyclin-D1表达水平虽降低,但组间比较差异无统计学意义(P>0.05);50和100μmol·L^(-1)红景天苷组细胞上清液中β-catenin、MMP-7和Cyclin-D1表达水平明显降低(P<0.05)。Western blotting法检测,模型对照组细胞中β-catenin蛋白表达水平明显高于正常对照组(P<0.01);12.5和25.0μmol·L^(-1)红景天苷组细胞中β-catenin蛋白表达水平虽低于模型对照组,但组间比较差异无统计学意义(P>0.05);50.0和100.0μmol·L^(-1)红景天苷组细胞中β-catenin蛋白表达水平低于模型对照组(P<0.05)。结论:红景天苷可抑制HFLS-RA的增殖,其控制RA的机制可能与Wnt/β-catenin信号通路的调控有关联。

Objective： To study the inhibitory effect of salidroside on the proliferation of fibroblast-like synoviocytes with rheumatoid arthritis in human （HFLS-RA） induced by tomor necrossi factor-α （TNF-α）, and to clarify the molecular mechanism of its control effect on rheumatoid arthritis （RA）. Methods： The HFLS-RA were cultured in vitro, then treated with TNF-a and different concentrations of salidroside. The cells were divided into normal control group （0 μg ~ L-1TNF-a）, model control group （10.0 μg ~ L-1TNF-α） and 12.5, 25.0, 50.0, and 100. 0 μmol · L-^1 salidroside groups （10.0 μg · L^-1 TNF-a ＋ salidroside）. The proliferation activity was detected by MTT mehthod; the expression levels of la-catenin, matrix metalloproteinase-7 （MMP-7）, and Cyclin- D1 in supernatant of the cells were detected by ELISA methodl the expression level of β-catenin protein in cells was detected by Western blotting method. Results： Compared with normal control group, the proliferation activity of the HFLS-RA in model control group was significantly increased （P《0.05） I compared with model control group, the proliferation activities of the HFLS-RA in 12.5 and 25.0 btmol · L ^-1 salidroside groups were decreased but there were no significant differences （P 〉 0. 05）, and the proliferation activities of the HFLS-RA in 50.0 and 100.0 btmol- L ^-1salidroside groups were significantly decreased （P〈 0.05）. Compared with normal control group, the expression levels of 13-catenin, MMP-7, and Cyclin-D1 in the supernatant of the cells in model control group were increased （P 〈0.05）. Compared with model control group, the expression levels of β-catenin, MMP-7, and Cyclin- D1 in the supernatant of the cells in 12.5 and 25.0 μmol · L-^1 salidroside groups were decreased, but there were no significant differences （P〉0.05）i the expression levels of β-catenin, MMP-7, and Cyclin- D1 in the supernatant of the cells in 50. 0 and 100.0 μmol · L^-1 salidroside groups were decreased （P〈0.05）. The Western blotting results showed that the expression level of β-catenin protein in the cell in model control group was higher than that in normal control group （P〈0.01） ; the expression levels of β-eatenin protein in the cells in 12.5 and 25.0 μmol · L-1 salidroside groups were lower than that in model control group, but there were no significant differences （P〉0.05） ; the expression levels of β-catenin protein in the cells in 50.0 and 100.0 μmol. L-1 salidroside groups were lower than that in model control group （P〈0.05）. Conclusion： Salidroside could inhibit the proliferation of HFLS-RA, and its control effect might be related to the regulation of Wnt/β- catenin single pathway.