为研制来源天然、毒副作用小的非酒精性脂肪肝病(non-alcoholic fatty liver disease,NAFLD)干预或缓解因子,该研究通过酶解方法制备豌豆低聚肽,检测其3-羟基-3-甲基戊二酰单酰辅酶A(3-hydroxy-3-methylglutaryl monocoenzyme A,HMG-CoA)还原酶抑制活性和体外NAFLD缓解作用及缓解机制研究。结果表明,100 mg/mL低聚肽抑制率达56.62%;人肝肿瘤(HepG2)细胞构建的NAFLD模型检测中,800 μg/mL的豌豆低聚肽能够显著减少细胞的内脂质沉积,且细胞内甘油三酯和总胆固醇含量也被显著降低。Western blot检测表明,豌豆低聚肽通过激活腺苷酸活化蛋白激酶磷酸化来抑制乙酰辅酶A羧化酶活性和甾醇调节元件结合蛋白2的表达,进一步上调肉碱棕榈酰转移酶 1的表达和下调3-羟基-3-甲基戊二酰辅酶A还原酶的表达,从而加强了脂肪酸的β-氧化,减弱了胆固醇的合成途径,最终降低了细胞内的脂质积累。
To develop intervention or mitigating factors for non-alcoholic fatty liver disease (NAFLD) of natural origin and with low toxicity and side effects, this study prepared pea oligopeptides by enzymatic hydrolysis and tested the 3-hydroxy-3-methylglutaryl monocoenzyme A (HMG-CoA) reductase inhibitory activity of pea oligopeptides and in vitro NAFLD mitigating effect and mitigating mechanism.Results showed that the inhibition rate of 100 mg/mL oligopeptide was 56.62%.At 800 μg/mL, pea oligopeptides could reduce intracellular lipid deposition significantly, and the intracellular triglyceride and total cholesterol contents were also significantly reduced in the NAFLD model assay constructed in HepG2 cells.Western blot assay shows that pea oligopeptides inhibit acetyl-CoA carboxylase (ACC)activity and sterol-regu-latory element binding protein 2 expression by activating adenosine 5′-monophosphate (AMP)-activated protein kinase(AMPK) phosphorylation, further up-regulation of carnitine acyl transferase I expression, and down-regulation of 3-hydroxy-3-methylglutaryl-CoA reductase expression. These enhance the β-oxidation of fatty acids and attenuate the cholesterol synthesis pathway. These results ultimately reduced intracellular lipid accumulation.
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