水飞蓟素改善棕榈酸引起的胰岛素抵抗及机制初步探讨  

Silymarin ameliorates palmitic acid induced insulin resistance and its mechanism

作　　者：孟然[1] 冯斌[1] 黄斌[1] 朱大龙[1] 

机构地区：[1]南京大学医学院附属南京鼓楼医院内分泌科,210008

出　　处：《中国糖尿病杂志》2017年第6期541-545,共5页Chinese Journal of Diabetes

基　　金：国家自然科学基金项目青年基金(81300651);南京市卫生局医学科技发展项目(YKK12057);江苏省六大人才高峰项目(2015-WSN-165)

摘　　要：目的评价水飞蓟素(Sil)对棕榈酸(PA)诱导HepG2细胞IR的作用并进一步揭示其机制。方法实验分为正常对照(NC)组,棕榈酸(PA)组,Sil干预低剂量(12.5μmol/L,L-Sil)组、中剂量(25μmol/L,M-Sil)组和高剂量(50μmol/L,H-Sil)组。PA诱导HepG2细胞IR,分别给予12.5,25,50μmol/L Sil作用6 h,测定葡萄糖消耗量,使用活性氧簇(ROS)试剂盒测定ROS活性水平。采用Western blot方法检测AKT以及还原型烟酰胺腺嘌呤二核苷酸磷酸(NADPH)氧化酶的NOX2、p67phox、p47phox蛋白表达水平。结果PA组与NC组比较,葡萄糖耗氧量降低[(41.2±6.1)%vs(100±12)%,P=0.0087],磷酸化AKT(p-AKT)蛋白表达量降低[(0.458±0.185)vs(1.00±0.13),P=0.0184],同时PA组ROS水平[(1567±189 vs(14±34)U/ml,P=0.0094]以及NAPDH氧化酶NOX2、p67、p47亚基的表达水平升高(P<0.05)。Sil各组与PA组比较可增加葡萄糖消耗量[(52.8±8.4)%vs(62.2±6.5)%vs(82.3±9.8)%],差异有统计学意义(P=0.041、0.039、0.0076);H-Sil组磷酸化AKT(p-AKT)蛋白表达量提高(0.793±0.249),与PA组比较,差异有统计学意义(P=0.0395);Sil可降低ROS水平[(1085±252)vs(851±184)vvs(405±35)],与PA组比较,差异有统计学意义(P=0.047、0.023、0.0015),以及降低NAPDH氧化酶NOX2、p67、p47亚基的表达水平(P<0.05),且作用强度随Sil剂量的增加而增强。结论Sil可能通过降低NADPH氧化酶活性降低氧化应激,从而改善HepG2细胞的IR。Objective To assess the impact of silymarin （Sil） on insulin resistance （IR） induced by palmitic acid （PA） in HepG2 cells and to explore the potential mechanisms. Methods HepG2 cells were randomly divided into three groups： normal group, PA induced IR group （PA group） and Sil groups （low, medium and high dose, respectively）. IR was induced by 200μmol/L PA in HepG2 cells. The IR model was incubated by 12. 5,25 and 50μmol/L Sil respectively for 6 hours. The changes in glucose consumption and the activity of ROS were measured by commercial kits. The protein levels of AKT and NADPH oxidase subunits （NOX2, p67phox and p47phox） were determined by Western-blot. Results Compared with control group, PA group significantly decreased glucose computation [（41.2 ± 6.1） % vs （ 100 ± 12）%,P〈0.01]and the activity of p-AKT [（0. 458±0. 185） vs （1.00±0.13）,P〈0. 01],and increased the level of ROS [（1567±189） vs （314±34）U/ml,P〈0.01] and the activities of NADPH oxidase （P〈0. 01,P〈0. 01 and P〈0.05, respectively）. In Sil treated groups, the glucose consumption was higher （52.8± 8.4） % in 12.5 μmol/L group, （62.2 ± 6.5） % in 25 μmol/L group and （82.3 ±9.8）% in 50 μmol/L group）, （P〈0.05, P〈0.05, P〈0.01 vs PA group）, the activity of p-AKT were higher in 50 μmol/L （0. 793±0. 249） （P〈0.01 vs PA group）, the levels of ROS were lower [（1085±252） in 12.5 μmol/L group, （851±184） in 25 μmol/L group and （405±35） in 50μmol/L group] （P〈0.05 ,P〈0.01, P〈0.01 vs PA group）, and the activities of NADPH oxidase were lower （P〈0.05 vs PA group）. The effects of Sil increased along with its concentration. Conclusion These novel data indicated that Sil may improve IR through alleviating oxidative stress by inhibiting NADPH oxidase expression in HepG2 cells.

关 键 词：水飞蓟素 氧化应激 胰岛素抵抗 NADPH氧化酶 活性氧簇 

分 类 号：R587.1[医药卫生—内分泌]