DHA和EPA调节3T3-L1脂肪细胞胰岛素抵抗及糖脂代谢的作用差异  

A COMPARATIVE STUDY ON DHA AND EPA IN IMPROVING INSULIN RESISTANCE AND GLYCOLIPID METABOLISM IN 3T3-L1 ADIPOCYTES

作　　者：谭伟烽 黄洁 李旭东 魏文婷 毛丽梅[1] TAN Wei-feng;HUANG Jie;LI Xu-dong;WEI Wen-ting;MAO Li-mei(Department of Nutrition and Food Hygiene,School of Public Health,Southern Medical University,Guangzhou 510515,China)

机构地区：[1]南方医科大学公共卫生学院营养与食品卫生学系,广州510515

出　　处：《营养学报》2022年第1期57-63,71,共8页Acta Nutrimenta Sinica

基　　金：国家自然科学基金(No.81773425;No.82073542)。

摘　　要：目的 探讨n-3多不饱和脂肪酸DHA和EPA调节3T3-L1脂肪细胞胰岛素抵抗(insulin resistance,IR)及糖脂代谢的剂量反应关系和作用差异。方法:用10ng/ml TNF-α处理分化成熟的3T3-L1脂肪细胞48h,诱导IR。使用Western blot检测胰岛素信号通路,葡萄糖氧化酶法(GOD-POD)法测定糖摄取,验证诱导IR效果。建模成功后,采用25、50、100、200、400μmol/L的DHA或EPA与TNF-α联合干预24h,同时设立对照组。干预后使用GOD-POD检测糖摄取量,油红O染色测定胞内甘油三酯(triglyceride,TG),PCR测定固醇调节元件结合蛋白-1c(SREBP-1c)、乙酰辅酶A羧化酶(ACC)、脂肪酸合酶(FAS)、脂肪甘油三酯脂肪酶(ATGL)、激素敏感性脂肪酶(HSL)mRNA表达,Western blot检测胰岛素信号通路蛋白表达。结果:TNF-α损伤胰岛素信号通路,降低胰岛素敏感性,IR建模成功。在糖代谢方面,25-400μmol/L的DHA和EPA均能改善TNF-α损伤的糖摄取,其中100和200μmol/L的效果最为显著,另外50、200、400μmol/L DHA改善效果优于同浓度EPA。脂代谢方面,各浓度DHA和EPA均增加胞内TG;适宜浓度DHA和EPA可增加促脂质合成相关基因表达,其中DHA效果更显著;除400μmol/L EPA外,各浓度DHA和EPA均能降低HSL表达。信号通路调节方面,适宜浓度的DHA和EPA可改善胰岛素底物1(IRS1)、磷脂酰肌醇-3激酶(PI3K)、磷酸化蛋白激酶B(p-Akt)和葡萄糖转运受体4(GLUT4)表达,其中100、200μmol/L的调节效果最为显著,且DHA优于EPA。结论:TNF-α干预可诱导脂肪细胞IR。DHA和EPA在适宜浓度可改善糖脂代谢,调节胰岛素信号通路,但DHA效果优于EPA。当两者浓度为100、200μmol/L时,调节效果最佳。[营养学报,2022,44(1):57-63,71]Objective To explore differences and dose-respones of DHA and EPA on the regulation of insulin resistance and glycolipid metabolism in 3T3-L1 adipocytes. Methods To induce cellular insulin resistance,differentiated 3T3-L1 adipocytes were treated with 10ng/mL TNF-α for 48 hours. After the model was built,3T3-L1 adipocytes were treated with 25, 50, 100, 200, 400μmol/L DHA or EPA combined with TNF-α for 24h, and a control group was established at the same time. After the intervention, GOD-POD was used to detect glucose uptake, oil red O staining to determine intracellular TG, PCR to determine the mRNA expression of SREBP-1c, ACC, FAS, HSL, ATGL, and Western blot to detect the expression of the insulin signaling pathway. Results Intervention with TNF-α affected the insulin signal pathway and decreased insulin sensitivity in adipocytes. 25-400μmol/L DHA and EPA improved the glucose uptake. Among them, 100 and 200μmol/L DHA and EPA exhibited the most significant effect. Moreover, at the same concentration(50,100 and 400 μg/ml), DHA was better in improving glucose uptake compared to EPA. All concentrations of DHA and EPA increased intracellular TG. However,DHA and EPA treatments at an appropriate concentration increased the expression of lipogenesis-related genes and DHA was more effective. Except for 400μmol/L EPA, all concentrations of DHA and EPA reduced HSL mRNA expression. Appropriate concentrations of DHA and EPA improved the expression of IRS1, PI3K, p-Akt, UT4. The improvement effects of DHA and EPA at the concentrations of 100 and 200μmol/L were more significant, and DHA was better than EPA. Conclusion Treatment with TNF-α induced 3T3-L1 adipocytes to develop insulin resistance. Appropriate concentrations of DHA and EPA improved glycolipid metabolism and insulin signaling pathway in IR adipocytes. At the concentration of 100 and 200 μmol/L, both DHA and EPA exhibited the most significant effect, and the improvement effect of DHA is stronger than EPA. [ACTA NUTRIMENTA SINICA, 2022, 44(1):57-63,71]

关 键 词：3T3-L1脂肪细胞 DHA EPA 胰岛素抵抗 糖脂代谢 

分 类 号：R151.2[医药卫生—营养与食品卫生学]