紫色悬钩子提取物对TNF-α诱导3T3-L1脂肪细胞胰岛素抵抗模型的影响  

The Effect of Rubus irritans Extract on TNF-α Induced 3T3-L1 Adipocyte Insulin Resistance Model

作　　者：刘悦 余松 韩永霞 铁芳芳 童丽[1,3] 杨芳 LIU Yue;YU Song;HAN Yongxia;TIE Fangfang;TONG Li;YANG Fang(Medical College,Qinghai University,Xining,Qinghai 810016,China;Northwest Institute of Plateau Biology,Chinese Academy of Sciences,Xining,Qinghai 810008,China;Qinghai Provincical Key Laboratory of Traditional Chinese Medicine Research for Glucolipid Metabolic Diseases,Xining,Qinghai 810001,China)

机构地区：[1]青海大学医学院,青海西宁810016 [2]中国科学院西北高原生物研究所,青海西宁810008 [3]青海省糖脂代谢疾病防控中医药重点实验室,青海西宁810001

出　　处：《今日药学》2022年第4期257-263,共7页Pharmacy Today

基　　金：青海省自然科学基金资助项目(2020-ZJC-942Q)。

摘　　要：目的利用肿瘤坏死因子-α(TNF-α)诱导培养建立及鉴定3T3-L1脂肪细胞胰.岛素抵抗模型,探讨紫色悬钩子提取物对胰岛素抵抗的影响。方法采用3-异丁基-1-甲基黄嘌呤(IBMX)、地塞米松(Dex)、胰.岛素联合诱导3T3-L1前脂肪细胞分化为成熟的脂肪细胞,分别与0,2,5,10,15,20 ng·mL^(-1)TNF-α共孵育24,48,72,96 h;MTT法检测紫色悬钩子提取物和TNF-α对3T3-L1脂肪细胞活力的影响;油红O染色观察细胞形态的变化;Western Blot检测Akt、p-Akt蛋白表达水平;流式细胞术检测细胞对2-NBDG的摄取。结果TNF-α浓度大于等于10 ng·mL^(-1),诱导时间大于等于72 h能够显著降低胰岛素刺激后p-Akt的表达(P<0.01),产生胰岛素抵抗;胰岛素刺激后,与模型组相比,紫色悬钩子提取物都能够不同程度提高细胞中p-Akt的表达和对2-NBDG的摄取能力,其中Fr.3作用最显著(P<0.05).结论TNF-α诱导培养可以成功建立胰岛素抵抗细胞模型,紫色悬钩子提取物具有改善胰岛素抵抗的作用。OBJECTIVE To establish and identify 3T3-L1 adipocyte insulin resistance model inducing by tumor necrosis factor-α(TNF-α),and to explore the effect of Rubus irritans extract on insulin resistance.METHODS 3-isobutyl-1-methylxanthine(IBMX),dexamethasone(Dex),and insulin were used to induce 3T3-L1 preadipocytes to differentiate into mature adipocytes,then they were incubated with 0,2,5,10,15,20 ng·mL^(-1) TNF-α for 24,48,72,96 h,respectively.MTT method was used to detect the effect of Rubus irritans extract and TNF-α on the viability of 3T3-L1 adipocytes.Oil red O staining was used to observe the cells Morphological changes.Western blot was used to detect Akt and p・Akt protein expression levels.Flow cytometry was used to detect cell uptake of 2-NBDG.RESULTS TNF-α with the concentration greater than or equal to 10 ng·mL^(-1),and the induction time longer than or equal to 72 h,which could significantly reduce the expression of p-Akt after insulin stimulation(P<0.01),resulting in insulin resistance.After insulin stimulation,compared with the model group,the Rubus irritans extract could increase the expression of p-Akt in cells and the uptake capacity of 2-NBDG to varying degrees,among which Fr.3 had the most significant effect(P<0.05).CONCLUSION TNF-αinduction culture can successfully establish insulin resistance cell model,and Rubus irritans extract can improve insulin resistance.

关 键 词：紫色悬钩子 3T3-L1脂肪细胞 胰岛素抵抗细胞模型 肿瘤坏死因子-Α 2-NBDG 

分 类 号：R966[医药卫生—药理学]