低氧下α-MEM、DMEM高糖培养基对HepG2细胞线粒体Tom20表达水平及脂肪沉积量的影响

Effects of α-MEM and DMEM on Tom20 expression and fat deposition in the mitochondria of HepG2 cells under hypoxia

目的探究低氧下α-MEM、DMEM高糖培养基对HepG2细胞线粒体膜蛋白Tom20表达水平及脂肪沉积量的影响。方法将HepG2细胞设为常氧α-MEM组(21%O_(2))、常氧DMEM组(21%O_(2))、低氧a-MEM组(1%O_(2))、低氧DMEM组(1%O_(2)),用低氧刺激72 h,采用细胞生长密度评估法检测低氧下高糖培养基对细胞增殖的影响,采用CCK-8法检测细胞活力的影响,采用Western blot法检测细胞线粒体膜蛋白标志分子Tom20表达水平,采用油红0染色法检测细胞脂肪沉积量。结果低氧下a-MEM、DMEM高糖培养基对细胞生长密度、细胞活力及Tom20表达水平存在影响:均降低(P<0.05),油红0染色着色区域较大;但高糖DMEM组细胞生长密度、细胞活力比对照常氧α-MEM组显著升高,Tom20表达水平升高(P<0.05),油红0染色区域较小。结论低氧显著抑制HepG2细胞线粒体膜蛋白Tom20的表达水平、增加脂肪沉积量;低氧下采用高糖DMEM培养显著降低HepG2细胞线粒体代谢能力,增加了脂肪沉积量。

Objective To explore the effects of α-MEM and DMEM on the expression of mitochondrial membrane protein Tom20 and fat deposition in HepG2 cells under hypoxia.Methods Divided the HepG2 cells as nor-moxia α-MEM group(21%O_(2)),normoxia DMEM group(21%O_(2)),hypoxia a-MEM group(1%O_(2))and hypoxia DMEM group(l%O_(2));the effect of high glucose medium on cell proliferation under hypoxia was detected by cell growth density assessment method after 72 hours of stimulation.The effects of cell viability were detected by CCK-8.The expression level of mitochondrial membrane protein marker Tom20 was detected by Western blot.The amount of cell fat deposition was detected by the Oil Red 0 Staining method.Results α-MEM and DMEM decreased the cell growth density,cell viability and Tom20 expression level(P<0.05)under hypoxia,and the staining area of Oil Red 0 was large;but compared with α-MEM group,the cell growth density and cell viability of DMEM group were significantly increased and the expression level of Tom20 was increased(P<0.05)with smaller staining area of Oil Red 0.Conclusion Hypoxia significantly inhibits the expression of mitochondrial membrane protein Tom20 and increases fat deposition in HepG2 cells;the utilization of DMEM culture significantly reduces the mitochondrial metabolism and increases the amount of fat deposition of HepG2 cells under hypoxia.