硫辛酸烟酸二联体拮抗丙烯醛诱导ARPE-19细胞凋亡的作用机制

摘要

Objective To investigate the regulations of Bax , Bcl-2 in the protection of lipoic acid-niacin diad in acrolein-induced apoptosis in ARPE-19 cells. Methods The ARPE-19 cells were cultured in medium containing 10% fetal bovine serum , at 37 ℃ with 5% CO2. The ARPE-19 was transferred to 6-well plate after reaching to 70% confluence. After starvation for 24 h , the cells in 6-well plates were divided into three groups , including the blank control group , the acrolein treatment group with 50 μmol/L acrolein for 24 h , and the protection group with 100 μmol/L lipoic acid-niacin diad for 24 h and with the acrolein for another 24 h. The apoptotic cells were detected by flow cytometry assay , and expressions of Bcl-2 , Bax protein were detected by Western Blot assay. Results The percentages of normal healthy cells were 94.8%, 60.98%, and 91.34% in the blank control group , 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group , respectively. The ratios of Bax/Bcl-2 protein expression were 0.293 9, 1.389 2, and 0.555 8 in the blank control group, 50 μmol/L acrolein group and 100 μmol/L diad contained of lipoic acid and niacin group, respectively. Conclusion The protective effect of lipoic acid-niacin diad on acrolein-induced apoptosis in ARPE-19 cell through promoting Bcl-2 expression and inhibiting Bax expression.%目的：探讨Bax、Bcl-2蛋白在硫辛酸烟酸二联体拮抗丙烯醛诱导的体外培养 ARPE-19细胞凋亡中可能的调控作用。方法：ARPE-19细胞置于含有10％体积分数胎牛血清的DMEM培养基，37℃、体积分数5％CO2培养箱中培养，待培养瓶底部ARPE-19细胞长至70％时，种6孔板，用不含血清的DMEM培养基培养24 h ，细胞换液。实验分为3组：空白对照组、丙烯醛组和硫辛酸烟酸二联体组。细胞流式检测各组ARPE-19细胞凋亡情况，Western blot检测Bax、Bcl-2蛋白的表达水平。结果：细胞流式显示正常细胞在空白对照组中为94．8％，丙烯醛组中为60．98％，硫辛酸烟酸二联体组中为91．34％。Western Blot显示空白对照组的Bax/Bcl-2蛋白表达的比值为0．2939，丙烯醛组的 Bax/Bcl-2蛋白表达的比值为1．3892，硫辛酸烟酸二联体组的Bax/Bcl-2蛋白表达的比值为0．5558。结论：硫辛酸烟酸二联体对ARPE-19细胞的保护作用机制是通过激活抗凋亡蛋白Bcl-2来抑制凋亡蛋白Bax的表达。