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目的观察水飞蓟宾对油酸诱导的非酒精性脂肪肝(NAFLD)模型脂代谢的影响。方法将细胞分为空白组、模型组(0.2 mmol·L~(-1)油酸)和3个剂量实验组(0.2 mmol·L~(-1)油酸+2.5,5.0,10.0μg·m L~(-1)水飞蓟宾)。以酶偶联比色法检测细胞三酰甘油(TG),以黄嘌呤氧化酶法测定超氧化物歧化酶(SOD),以硫代巴比妥酸法测定丙二醛(MDA),以酶促法测定谷胱甘肽过氧化物酶(GSH-Px),以酶联免疫吸附法测定肿瘤坏死因子-α(TNF-α)水平,以实时荧光定量PCR法检测细胞腺苷酸活化蛋白激酶(AMPK)、去乙酰化酶(Sir T3)、乙酰辅酶A羧化酶2(ACC 2)及肉碱棕榈酰转移酶~(-1)A(CPT~(-1)A)的mRNA表达水平,以油红O染色观察细胞内脂滴沉积变化。结果高剂量实验组TG水平为(164.74±53.94)mg·g~(-1),与模型组的(500.05±152.01)mg·g~(-1)相比显著降低(P<0.01);与模型组SOD水平(4.96±1.39)U·mg~(-1)比较,高剂量实验组SOD水平(20.37±11.16)U·mg~(-1)显著增高(P<0.01)。与模型组MDA、TNF-α水平(0.20±0.03)U·mg~(-1)、(32.03±8.04)pg·m L~(-1)比较,高剂量实验组的MDA、TNF-α水平(0.10±0.02)U·mg~(-1)、(15.00±0.53)pg·m L~(-1)显著降低(P<0.01,P<0.05)。与模型组比较,高剂量实验组的Ampk、Sir T3、CPT~(-1)A mRNA表达水平显著增加(P<0.01),而ACC2 mRNA表达显著降低(P<0.01)。结论水飞蓟宾有效降低油酸诱导的NAFLD模型肝细胞TG水平,可能与其抗氧化、调节脂代谢相关基因表达水平相关。
Objective To observe the effect of silibinin on lipid metabolism in oleic acid-induced non-alcoholic fatty liver disease (NAFLD) model. Methods The cells were divided into blank group, model group (0.2 mmol·L -1 oleic acid) and three dose groups (0.2 mmol·L -1 oleic acid + 2.5, 5.0 and 10.0 μg · m -1) L ~ (-1) silybin). Cell triglyceride (TG) was detected by enzyme-linked immunosorbent assay (ELISA), superoxide dismutase (SOD) was measured by xanthine oxidase method, malondialdehyde (MDA) was measured by thiobarbituric acid method, The level of tumor necrosis factor-α (TNF-α) was determined by the method of enzyme-linked immunosorbent assay (GSH-Px) and the expression of adenosine triphosphate kinase The expressions of AMPK, Sir T3, ACC 2 and CPT -1 (A) The changes of lipid droplets in cells were observed by oil red O staining. Results The level of TG in high dose group was (164.74 ± 53.94) mg · g ~ (-1), which was significantly lower than that in model group (500.05 ± 152.01 mg · g ~ (-1)) (P <0.01) Compared with the model group, SOD level (4.96 ± 1.39) U · mg ~ (-1) was significantly increased (P <0.01) in high dose group compared with 20.37 ± 11.16 U · mg ~ (-1) group. Compared with the model group, MDA and TNF-αlevel of experimental group were significantly higher than that of model group (0.20 ± 0.03) U · mg -1, (32.03 ± 8.04) pg · m L -1 (0.10 ± 0.02) U · mg ~ (-1) and (15.00 ± 0.53) pg · m L ~ (-1) significantly decreased (P <0.01, P <0.05). Compared with the model group, the expression of Ampk, Sir T3 and CPT ~ (-1) A mRNA in the high dose group increased significantly (P <0.01), while the expression of ACC2 mRNA decreased significantly (P <0.01). Conclusion Silybin can effectively reduce the triglyceride level in hepatic cells induced by oleic acid in NAFLD model, which may be related to the anti-oxidation and regulation of lipid metabolism-related gene expression.