薯蓣皂苷对TNF-α诱导的CIA大鼠成纤维样滑膜细胞COX表达的影响

Effect of Diosgenin on TNF-α-Induced COX Expression in Fibroblast-Like Synoviocytes of CIA Rats

目的通过研究薯蓣皂苷(Dio)对肿瘤坏死因子α(TNF-α)诱导的CIA大鼠成纤维样滑膜(FLS)细胞环氧合酶1(COX1)、环氧合酶2(COX2)表达的影响,探索Dio对TNF-α诱导的CIA大鼠FLS细胞的治疗作用。方法应用TNF-α刺激CIA大鼠FLS细胞,采用MTT法检测FLS细胞活性。对TNF-α刺激的CIA大鼠FLS细胞给予不同浓度(1、2、3、4μg·mL^-1)的Dio处理24 h,采用Q-PCR检测FLS细胞COX1和COX2 mRNA的表达,Western blot检测细胞COX2蛋白的表达。结果浓度为3μg·mL^-1的Dio作用于FLS细胞,存活率接近50%,因此用药组浓度确定为1、2、3、4μg·mL^-1。TNF-α诱导后,COX1 mRNA、COX2 mRNA和COX2蛋白表达均显著升高(P<0.01)。与TNF-α组相比,Dio≤3μg·mL^-1时,均能不同程度地降低细胞中COX2 mRNA和蛋白的表达(P<0.05或P<0.01),Dio≥3μg·mL^-1能显著抑制COX1 mRNA的表达(P<0.05或P<0.01)。结论Dio通过抑制COX2 mRNA和蛋白的表达,达到治疗TNF-α诱导的CIA大鼠FLS细胞的作用。

Objective To investigate the effect of diosgenin(Dio)on tumor necrosis factor-α(TNF-α)-induced expression of cyclooxygenase 1(COX1)and cyclooxygenase 2(COX2)in fibroblast-like synoviocytes(FLSs)of CIA rats,and to explore the therapeutic action of Dio in TNF-α-treated FLSs.Methods MTT assay was used to measure the activity of TNF-α-stimulated CIA rat FLSs.After cells were treated with Dio(1,2,3,and 4μg·mL^-1)for 24 hours,the expression of COX1 and COX2 mRNA was detected by Q-PCR and the expression of COX2 protein was determined by Western blot.Results Cell survival rate was close to 50%when the concentration of Dio was 3μg·mL^-1.Therefore,the concentrations of Dio were selected as 1,2,3 and 4μg·mL^-1.The expression of COX1 and COX2 mRNA and COX2 protein significantly increased after TNF-αinduction(P<0.01).Compared with TNF-αgroup,treatment with Dio at≤3μg·mL^-1 reduced the expression of COX2 mRNA and protein in FLSs(P<0.05 or P<0.01).Furthermore,Dio at≥3μg·mL^-1 inhibited COX1 mRNA expression in FLSs(P<0.05 or P<0.01).Conclusion Dio can treat TNF-α-induced CIA rat FLSs through inhibiting COX2 mRNA and protein expression.