辣木叶水提物对油酸钠-钠棕榈酸酯诱导的HepG2细胞脂质代谢及抗氧化能力的影响

Effects of Moringa oleifera leaf aqueous extract on lipid metabolism and antioxidant capacity in HepG2 cells induced by sodium oleate-sodium palmitate

目的探讨辣木叶水提物(MOLAE)对脂肪堆积人HepG2细胞的调节作用。方法制备MOLAE冻干粉;通过CCK-8法检测油酸钠-钠棕榈酸酯(O-P)对HepG2细胞活力的影响、油红O染色检测O-P对细胞脂质堆积的影响、试剂盒法检测O-P对细胞三酰甘油(TG)、总胆固醇(TC)水平的影响,筛选O-P诱导HepG2细胞脂质代谢异常模型的作用浓度及时间;CCK-8法检测辛伐他汀、MOLAE对HepG2细胞活力的影响,筛选安全作用浓度;设立对照组、模型组、辛伐他汀(阳性药,15μmol·L^(−1))组和MOLAE(3.125、6.250、12.500、25.000、50.000μg·mL^(−1))组,除对照组外,其他各组均给予0.4-0.2 mmol·L^(−1)的O-P诱导细胞脂质沉积,诱导3 h后开始给药,干预24 h后,CCK-8法检测细胞活性,油红O染色观察细胞中脂滴形成情况,试剂盒法测定细胞中TG、TC、谷胱甘肽(GSH)、超氧化物歧化酶(SOD)及丙二醛(MDA)水平。结果确定造模方式为:0.4-0.2 mmol·L^(−1)的O-P作用HepG2细胞3 h后开始给药;10、15μmol·L^(−1)的辛伐他汀和3.125~100.000μg·mL^(−1)的MOLAE作用于HepG2细胞24、48 h后,不影响细胞活力。与模型组比较,不同浓度的MOLAE(3.125、6.250、12.500、25.000、50.000μg·mL^(−1))均能降低TG、TC、MDA水平(P<0.05、0.01),显著升高GSH、SOD水平(P<0.05、0.01)。油红O染色结果表明,辛伐他汀组和MOLAE组(12.5、25.0μg·mL^(−1))脂滴堆积现象均较模型组有明显改善。结论MOLAE能够降低O-P诱导的HepG2细胞中TG、TC水平,提高GSH、SOD水平和降低MDA水平,减少细胞中脂质堆积的现象。

Objective To investigate the regulating effects of Moringa oleifera leaf aqueous extract(MOLAE)on lipid abnormality in human HepG2 cells.Methods Detect the effect of sodium oleate-sodium palmitate(O-P)on HepG2 cell viability using the CCK-8 method,the effect of O-P on cell lipid accumulation using oil red O staining,and the effect of O-P on cell triglyceride(TG)and total cholesterol(TC)levels using the kit method.Screen the concentration and time of O-P induced abnormal lipid metabolism model in HepG2 cells.The CCK-8 method was used to detect the effects of simvastatin and MOLAE on the viability of HepG2 cells,and to screen for safe concentrations of action.Establish control group,model group,and simvastatin(positive drug,15μmol·L^(−1))group and the MOLAE(3.125,6.250,12.500,25.000,50.000μg·L^(−1))group,except for the control group,were all given O-P of 0.4-0.2 mmol·L^(−1)to induce cell lipid deposition.After induction for 3 h,the drug was administered.After 24 hours of intervention,cell activity was detected using CCK-8 method,lipid droplet formation was observed using oil red O staining,and the levels of TG,TC,superoxide dismutase(SOD),malondialdehyde(MDA),and glutathione(GSH)in cells were measured using a kit method.Results The modeling method was determined as follows:Administration of O-P with a concentration of 0.4-0.2 mmol·L^(−1)was initiated on HepG2 cells for 3 h.Simvastatin with 10 and 15μmol·L^(−1)and MOLAE of 3.125—100.000μg·mL^(−1)did not affect cell viability after 24 and 48 hours of action on HepG2 cells.Compared with the model group,different concentrations of MOLAE(3.125,6.250,12.500,25.000,50.000μg·mL^(−1))can reduce the levels of TG,TC,and MDA(P<0.05,0.01),and significantly increase the levels of GSH and SOD(P<0.05,0.01).The oil red O staining results showed that the accumulation of lipid droplets in the simvastatin group and MOLAE(12.5,25.0μg·mL^(−1))group was significantly improved compared with model group.Conclusion MOLAE can reduce TG and TC levels in model group,increase GSH and SOD levels and reduce MDA levels,reduce lipid accumulation in cells and prevent lipid damage.