人参皂苷通过Aldolase/AMPK/PINK1信号通路减轻油酸诱导的HL-7702细胞损伤

目的: 研究人参皂苷对油酸(OA)诱导人肝HL-7702细胞损伤的保护作用,及其对Aldolase/AMPK/PINK1信号通路的影响。方法: 体外培养HL-7702细胞,并将其分为对照组(C)、油酸刺激组(OA)、OA+人参皂苷组(OA+Rg1)、OA+Compound C组(OA+CC)、OA+Compound C+人参皂苷组(OA+CC+Rg1)。CCK8法和Hoechst染色法检测HL-7702细胞增殖活性,流式细胞术检测HL-7702细胞凋亡率和线粒体膜电位,ATP测定分析HL-7702细胞中ATP含量,Western blot分析HL-7702细胞中Cleaved caspase-3、PINK1、MFN2、Aldolase和p-AMPK蛋白表达水平。结果: 与C组比较,OA组细胞增殖活力明显降低(P<0.05),细胞凋亡率和促凋亡蛋白Cleaved caspase-3表达水平明显升高(P<0.05),ATP含量、线粒体膜电位(TMRE)和线粒体自噬蛋白PINK1表达水平明显降低(P<0.05);与OA组比较,OA+Rg1组细胞增殖活力明显升高(P<0.05),细胞凋亡率和Cleaved caspase-3蛋白表达水平明显降低(P<0.05),ATP含量、TMRE值、Aldolase、PINK1和p-AMPK蛋白表达水平明显升高(P<0.05);与OA+Rg1组比较,OA+CC+Rg1组HL-7702细胞增殖活力及AMPK磷酸化水平明显降低(P<0.05);Aldolase基因沉默后,Rg1不能增高细胞中PINK1和p-AMPK蛋白表达及细胞的增殖活性。结论: 人参皂苷可通过调节Aldolase/AMPK/PINK1信号通路活性改善油酸诱导的人肝HL-7702细胞损伤。

Ginsenoside Rg1 protects HL-7702 cells against oleic acid-induced injury via aldolase/AMPK/PINK1 signalling

AIM: To study the protective effects of ginsenoside Rg1 on HL-7702 cells injury induce by oleic acid (OA), and investigate its role in aldolase/AMPK/PINK1 signalling. METHODS: HL-7702 cells were cultured in vitro and divided into five groups: control group (C), oleic acid group (OA), OA+ginsenoside (Rg1) group, OA+compound C (CC) group, and OA+CC+Rg1 group. The viability of HL-7702 cells was determined by CCK8 assay and Hoechst staining. The apoptosis and mitochondrial membrane potentials of HL-7702 cells were measured using flow cytometry. ATP content in HL-7702 cells was observed. Western blot was used to detect the expression levels of Cleaved caspase-3, PINK1, MFN2, Aldolase and p-AMPK in HL-7702 cells. RESULTS: Compared with C group, the viability of cells in OA group was significantly decreased (P<0.05), the apoptotic rate and Cleaved caspase-3 expression were greatly increased (P<0.05), ATP level, mitochondrial membrane potentials (TMRE) and PINK1 expression were significantly decreased (P<0.05). Compared with OA group, the viability of cells in OA+Rg1 group was significantly increased (P<0.05), the apoptotic rate and Cleaved caspase-3 expression were greatly decreased (P<0.05), ATP level, mitochondrial membrane potentials (TMRE) and PINK1 expression were significantly increased (P<0.05). Compared with OA+Rg1 group, the viability of cells and p-AMPK expression level in OA+CC+Rg1 group was significantly decreased (P<0.05). Reducing the expression of aldolase in cells inhibited Rg1?s actions on PINK1 and p-AMPK and cell viability. CONCLUSION: Ginsenoside Rg1 can improve the injury of HL-7702 cells via regulating aldolase/AMPK/PINK1 signaling pathway.