胡桃醌对类风湿关节炎关节成纤维样滑膜细胞增殖和凋亡的影响

Effects of juglone on cell proliferation and apoptosis in rheumatoid arthritis fibroblast-like synoviocytes

目的探讨胡桃醌对类风湿关节炎关节成纤维样滑膜细胞(RA-FLSs)增殖和凋亡的影响。方法不同浓度胡桃醌(0、5、10、20、30、40、50μmol/L)孵育RA-FLSs 24 h,胡桃醌(30μmol/L)分别和不同终质量浓度Akt激动剂SC79(0、0.1、0.5、0.8、1.0、1.5、1.8μg/mL)同时孵育RA-FLSs 24h。BrdU和MTT法分别检测RA-FLSs的增殖能力和细胞活力,Annexin-V FITC/PI法和caspase-3活性检测试剂盒检测RA-FLSs凋亡,Western blot法检测Akt磷酸化和p21蛋白表达。结果与0μmol/L胡桃醌比较,胡桃醌(30、40、50μmol/L)显著抑制RA-FLSs细胞活力和增殖能力,促进其凋亡和caspase-3活性(P<0.01);30μmol/L胡桃醌能显著抑制Akt磷酸化,促进p21蛋白表达(P<0.01)。与30μmol/L胡桃醌比较,SC79(1.0μg/mL)与30μmol/L胡桃醌共同处理时,p21蛋白表达显著下降,并能促进RA-FLSs细胞凋亡,抑制其增殖(P<0.01)。结论胡桃醌可通过调节Akt磷酸化来调控p21表达,进而调节RA-FLSs增殖和凋亡过程,为类风湿关节炎的治疗提供价值。

AIM To investigate the effects of juglone on cell proliferation and apoptosis in rheumatoid arthritis fibroblast-like synoviocytes(RA-FLSs).METHODS RA-FLSs treated with 24 h incubation with different final concentrations of juglone(0, 5, 10, 20, 30, 40, 50 μmol/L), and RA-FLSs simultaneously incubated with juglone(30 μmol/L) and different final concentrations of Akt agonist SC79(0, 0.1, 0.5, 0.8, 1.0, 1.5 or 1.8 μg/mL) for 24 h, were subjected to test of their proliferation and cell viability by BrdU and MTT assays, respectively. Their apoptosis was measured by Annexin-V FITC/PI method and caspase-3 activity detection kit, and their Akt phosphorylation level and p21 protein expression were detected by Western blot.RESULTS In contrast to 0 μmol/L juglone, juglone at concentrations of 30, 40, and 50 μmol/L significantly inhibited the cell viability and proliferation of RA-FLSs, and promoted their apoptosis and caspase-3 activity(P<0.01);30 μmol/L juglone significantly inhibited Akt phosphorylation and promoted p21 protein expression(P<0.01). Compared with the single treatment of 30 μmol/L juglone, co-treatment of it with 1.0 μg/mL SC79 brought forth significant decrease of p21 protein expression, promotion of the apoptosis, and inhibition of the proliferation as well in RA-FLSs(P<0.01).CONCLUSION Regulating the expression of p21 by controlling the phosphorylation level of Akt, thereby modulating the proliferation and apoptosis of RA-FLSs, juglone highlights the treatment of rheumatoid arthritis.