摘要
目的:研究沙棘多糖对胰岛素抵抗HepG2细胞氧化应激的保护作用与机制。方法:采用水提醇沉法对沙棘多糖进行提取,并用苯酚硫酸法测定多糖含量。CCK-8法测定不同浓度沙棘多糖对HepG2细胞活力的影响,用含5×10^(-8)mol/L胰岛素的培养基诱导HepG2细胞24 h,建立HepG2细胞胰岛素抵抗模型。采用试剂盒测定葡萄糖消耗量以及糖原相对含量,并测定超氧化物歧化酶(SOD)、丙二醛(MDA)含量,Western Blot法测定氧化应激相关蛋白的表达。结果:沙棘多糖含量为88.46%,在沙棘多糖质量浓度达到800μg/mL时,细胞活力出现显著下降(P<0.05),后续安全剂量选择400μg/mL。经沙棘多糖处理后的葡萄糖消耗量以及糖原相对含量均显著提高(P<0.05),SOD水平达到(79.31±2.16)U/mg,MDA水平达到(2.15±0.12)nmol/mg。沙棘多糖处理后显著提高Nrf2和HO-1的表达水平(P<0.05),显著降低Keap1的表达水平(P<0.05)。结论:沙棘多糖能够改善胰岛素抵抗细胞模型异常糖代谢情况和氧化应激水平,并通过调控Nrf2/Keap1/HO-1通路起到改善胰岛素抵抗的作用。
Objective:The protective effect and mechanism of seabuckthorn polysaccharides on insulin-resistant HepG2 cells against oxidative stress was investigated.Methods:The seabuckthorn polysaccharide was extracted by water extraction and alcohol precipitation method,and the polysaccharide content was determined by the phenol sulfuric acid method.The CCK-8 method was used to determine the effects of different concentrations of sea-buckthorn polysaccharides on the viability of HepG2 cells.HepG2 cells were induced with a medium containing 5×10^(-8) mol/L insulin for 24 hours to establish a HepG2 cell insulin resistance model.A kit was used to determine glucose consumption and relative glycogen content,and to determine the content of superoxide dismutase(SOD)and malondialdehyde(MDA).Western Blot method was used to determine the expression of oxidative stress-related proteins.Results:The content of sea-buckthorn polysaccharide was 88.46%.When the concentration of sea-buckthorn polysaccharide reached 800μg/mL,the cell viability decreased significantly(P<0.05).The subsequent safe dose was 400μg/mL.Glucose consumption and relative glycogen content after seabuckthorn polysaccharide treatment were significantly increased(P<0.05),SOD level reached(79.31±2.16)U/mg,MDA level reached(2.15±0.12)nmol/mg.After seabuckthorn polysaccharide treatment,the expression level of Nrf2 and HO-1 was significantly increased(P<0.05),and the expression level of Keap1 was significantly reduced(P<0.05).Conclusion:Hippophae rhamnoides polysaccharide can improve the abnormal glucose metabolism and oxidative stress level of insulin resistance cell model,and can improve insulin resistance by regulating the Nrf2/Keap1/HO-1 pathway.
作者
王秋丹
赵凯迪
林长青
WANG Qiu-dan;ZHAO Kai-di;LIN Chang-qing(Department of Traditional Chinese Medicine,School of Medicine,Yanbian University,Yanji,Jilin 133000,China)
出处
《食品与机械》
北大核心
2022年第3期167-172,共6页
Food and Machinery
基金
吉林省重点科技攻关项目(编号:20170204023YY)。
作者简介
王秋丹,女,延边大学在读硕士研究生;通信作者:林长青(1970-),男,延边大学副教授,博士。E-mail:lincq0608@163.com。
