鸡树条荚蒾果多酚改善胰岛素抵抗HepG2细胞的糖代谢效应

Improvement of insulin resistant HepG2 cells glycometabolism effect by polyphenol from the fruit of Viburnum sargentii

【目的】探讨鸡树条荚蒾果多酚对HepG2细胞胰岛素抵抗(IR)模型的改善作用,以评价其降血糖活性。【方法】本研究采用高浓度胰岛素诱导,建立体外IR模型,并进行模型稳定性(细胞活性法)及可靠性(Z因子法)评估。试验设空白组、IR模型组、阳性对照(二甲双胍)组和荚蒾果多酚组,MTT法检测细胞活性;葡萄糖氧化酶法检测培养液葡萄糖含量,计算葡萄糖消耗量;蒽酮法测定糖原含量;比色法检测细胞己糖激酶(HK)、丙酮酸激酶(PK)活性,酶联免疫吸附试验(ELISA)检测细胞磷酸烯醇式丙酮酸激酶(PEPCK)、葡萄糖六磷酸酶(G6PC)活性。【结果】10-6 mol/L的胰岛素诱导处理HepG2细胞24 h是产生胰岛素抵抗模型的最适条件,且IR模型在12~36 h内有较高的稳定性和可靠性。0.10~1.00 mg/mL荚蒾果多酚组的葡萄糖消耗量显著高于模型组(P<0.05),24 h、0.50 mg/mL组的糖消耗量最高,为(3.49±0.11)mmol/L,消耗率可达88.81%(P<0.01)。与模型组对比,荚蒾果多酚可提高糖原的含量33.65%(P<0.01),HK、PK活性可分别提高43.36%(P<0.05)、48.41%(P<0.01),对G6PC、PEPCK活性抑制率为分别为22.86%(P<0.01)、17.33%(P<0.05)。【结论】鸡树条荚蒾果多酚可提高IR-HepG2细胞的HK、PK活性,加快糖酵解,增加糖原含量;抑制G6PC、PEPCK活性,从而减少细胞内源性葡萄糖的产生。所以鸡树条荚蒾果多酚对胰岛素抵抗细胞的治疗具有一定效果。

[Objective]This study investigated the improvement of the polyphenol from the fruit of Viburnum sargentii(PVSK)on insulin resistance(IR)model of HepG2 cells,and to evaluate its hypoglycemic activity.[Method]High concentration insulin was used to induce in vitro IR model,and the stability(cell activity method)and reliability(Z factor method)of the model were evaluated.Blank group,IR model group,positive control group(metformin)and PVSK group were set up.Cell viability was detected by MTT assay,glucose content in culture medium was detected by Glucose oxidase method and the consumption of glucose was clculated;glycogen content was determined by anthrone method;the activity of hexokinase(HK)and pyruvate kinase(PK)were determined by colorimetric method.Enzyme-linked immunosorbent assay(ELISA)was used to detect the activity of phosphoenolpyruvate kinase(PEPCK)and glucose hexaphosphatase(G6PC).[Result]Insulin induction treatment of HepG2 cells at 10-6 mol/L for 24 hours was the best condition for the production of insulin resistance model,and the IR model had high stability and reliability within 12~36 hours.0.10~1.00 mg/mL PVSK polyphenol group had significantly higher glucose consumption than model group(P<0.05).The highest glucose consumption was(3.49±0.11)mmol/L in 24 h and 0.50 mg/mL groups,with a consumption rate of 88.81%(P<0.01).Comparedwith model group,PVSK polyphenols could increase glycogen content by 33.65%(P<0.01),HK and PK activity by 43.36%(P<0.05)and 48.41%(P<0.01),respectively.The inhibition rates of G6PC and PEPCK activity were 22.86%(P<0.01)and 17.33%(P<0.05),respectively.[Conclusion]PVSK could increase HK and PK activities of IR-HepG2 cells,accelerate glycolysis and increase glycogen content.Inhibiting the activity of G6PC and PEPCK can reduce the production of endogenous glucose.Therefore,the polyphenols of PVSK have a certain effect on the treatment of insulin resistance cells.