白芥子对类风湿性关节炎滑膜细胞增殖、凋亡的影响

Effects of semen brassicae albae on proliferation and apoptosis of rheumatoid arthritis synoviocytes

目的 研究白芥子对类风湿性关节炎(RA)滑膜细胞增殖、凋亡的影响,并分析相关机制。方法 取对数生长期RA成纤维样滑膜细胞(RA-FLS)细胞,随机分为空白组、阳性对照组和低、中、高剂量实验组。空白组不作任何处理;低、中、高剂量实验组分别用含20、40、80μg·mL^(-1)白芥子提取物的DMEM培养基培养;阳性对照组用含5 mg·L^(-1)雷公藤多苷的DMEM培养基培养。5组均培养48 h。用CCK-8法、Annexin V-FITC/PI双染法分别检测RA-FLS细胞的存活率、凋亡率,用蛋白质印迹法检测增殖、凋亡相关蛋白的表达水平。结果 中、高剂量实验组和空白组、阳性对照组的细胞存活率分别为(71.35±10.86)%、(58.56±17.92)%、(99.12±0.38)%和(56.21±15.11)%,凋亡率分别为(28.98±4.35)%、(39.12±5.85)%、(8.96±1.36)%和(41.21±6.01)%,PCNA蛋白相对表达水平分别为0.87±0.13、0.50±0.08、1.30±0.19和0.49±0.09,Bcl-2蛋白相对表达水平分别为1.20±0.14、1.35±0.19、0.45±0.06和1.31±0.21,Bax蛋白相对表达水平分别为0.65±0.09、0.27±0.04、1.14±0.15和0.29±0.06。中、高剂量实验组及阳性对照组的上述指标与空白组比较,差异均有统计学意义(均P<0.05)。结论 白芥子可抑制RA-FLS细胞增殖并诱导细胞凋亡,其作用机制可能与调节增殖、凋亡相关蛋白表达有关。

Objective To investigate the effects of semen brassicae albae on the proliferation and apoptosis of rheumatoid arthritis(RA)synoviocytes,and to analyze the related mechanisms.Methods Logarithmic RA fibroblastic synovial cells(RA-FLS)were randomly divided into blank group,experimental-L,M,H groups and positive control group.Blank group did not make any treatment.ExperimentalL,M,H groups were cultured using DMEM medium containing 20,40,80μg·mL^(-1)of semen brassicae albae extract,respectively.Positive control group was cultured at DMEM medium containing 5 mg·L^(-1)of Tripterygium glycosides.Five groups were cultured for 48 h.The survival rates of RA-FLS cells were detected by CCK-8 method,the apoptosis rates of RA-FLS cells were detected by Annexin V-FITC/PI double staining method,and the expression levels of proliferation and apoptosis-related proteins were detected by Western blotting.Results The cell survival rates of experimental-M,-H groups,blank group and positive control group were(71.35±10.86)%,(58.56±17.92)%,(99.12±0.38)%and(56.21±15.11)%;the apoptosis rates were(28.98±4.35)%,(39.12±5.85)%,(8.96±1.36)%and(41.21±6.01)%;the expression levels of PCNA protein were 0.87±0.13,0.50±0.08,1.30±0.19 and 0.49±0.09;the expression levels of Bcl-2 protein were 1.20±0.14,1.35±0.19,0.45±0.06 and 1.31±0.21;the expression levels of Bax protein were 0.65±0.09,0.27±0.04,1.14±0.15 and 0.29±0.06,respectively.Compared with blank group,the differences of above indexes of experimental-M,-H groups and positive control group were statistically significant(all P<0.05).Conclusion Semen brassicae albae can inhibit the proliferation of RA-FLS cells and induce cell apoptosis,and its mechanism may be related to regulating the expression of proliferation and apoptosis related proteins.