受体相互作用蛋白140对吡格列酮改善胰岛β细胞糖脂毒性损伤的影响

发布时间：2018-01-13 19:02

  本文关键词：受体相互作用蛋白140对吡格列酮改善胰岛β细胞糖脂毒性损伤的影响　出处：《中国糖尿病杂志》2016年12期 　论文类型：期刊论文

  更多相关文章： 受体相互作用蛋白 吡格列酮 胰岛β细胞 糖脂毒性

【摘要】：目的探讨过氧化物酶增殖活化受体-γ(PPAR-γ)激动剂吡格列酮对胰岛β细胞糖脂毒性损伤的保护及受体相互作用蛋白140(RIP140)在其中的介导机制。方法将胰岛β细胞株MIN6细胞分为NC组、高糖高脂组、吡格列酮干预组。稳定过表达RIP140的MIN6细胞(O-RIP140-MIN6)和过表达绿色荧光蛋白(GFP)的MIN6细胞(GFP-MIN6)。分别予高糖高脂(25 rmmol/L葡萄糖+500μmol/L棕榈酸)和/或10/μmol/L吡格列酮干预。利用MTT分别检测各组细胞增殖率、流式细胞仪检测凋亡率、RT-PCR检测RIP140 mRNA、Western blot检测B淋巴细胞瘤-2(Bcl-2)的表达、硫代巴比妥酸法检测丙二醛(MDA)水平及黄嘌呤氧化酶法检测超氧化合物歧化酶(SOD)含量。结果 NC组、高糖高脂组及吡格列酮干预组MTT吸光值分别为:24 h(1.80±0.04)、(0.95±0.04)及(0.97±0.03);48 h(2.70±0.11)、(1.04±0.06)及(1.30±0.03)。NC组与高糖高脂组比较,差异有统计学意义(24 h:t=25.94,P0.01,48 h:t=24.00,P0.01)。高糖高脂组与吡格列酮干预组比较,差异有统计学意义(48 h:t=9.37,P0.01)。各组间的24 h凋亡率分别为(2.93±0.66)%、(48.08±3.95)%(vs NC组,t=19.54,P0.01)及(31.38±3.92)%(vs高糖高脂组,t=5.20,P0.01)。Bcl-2相对表达量分别为(1.14±0.06)、(0.42±0.02)(vs NC组,t=20.52,P0.01)及(0.86±0.04)(vs高糖高脂组,t=17.71,P0.01)。RIP140表达量分别为(1.13±0.11)、(2.34±0.21)(vs NC组,t=9.69,P0.01)及(1.63±0.13)(vs高糖高脂组(t=5.03,P0.01);高糖高脂组与NC组比较,MDA[(10.13±0.47vs(5.00±0.26)nmol/mg,t=16.57,P0.01]、SOD[(5.15±1.07)协(12.25±1.25)nmol/mg,t=7.51,P0.01]比较,差异均有统计学意义。高糖高脂组与吡格列酮干预组比较,MDA[(10.13±0.47)vs(7.83±0.36)nmol/mg,t=6.77,P0.01]、SOD[(5.15±1.07)v5(8.74±0.59)nmol/mg,t=5.16,P0.01)差异有统计学意义。O-RIP140-MIN6和GFP-MIN6细胞分别给予高糖高脂及吡格列酮处理后,两组MTT吸光值:24 h(1.04±0.07)vs(1.40±0.16)(t=5.01,P0.01),48 h(1.16±0.13)vs(1.98±0.14)(t=10.73,P0.01)。凋亡率为(41.95±4.88)%vs(31.26±2.86)%(t=2.97,P0.05)、Bcl-2相对表达为(0.22±0.04)vs(0.76±0.03)(t=21.54,P0.01),SOD为(7.53±0.71)vs(9.62±0.43)nmol/mg(t=4.36,P0.05),MDA为(10.23±0.28)vs(8.15±0.38)nmol/mg(t=7.63,P0.01)。结论高糖高脂促进胰岛β细胞损伤,吡格列酮通过下调RIP140表达来抑制高糖高脂对胰岛β细胞的损伤。
[Abstract]:Objective to investigate the peroxisome proliferator activated receptor gamma (PPAR- gamma) receptor agonist pioglitazone and protection of pancreatic islet beta cell glucolipotoxicity injury interacting protein 140 (RIP140) in which the dielectric guide mechanism. Methods MIN6 cells of islet beta cells were divided into NC group, high-fat group, pioglitazone intervention group is stable. Over expression of RIP140 MIN6 cells (O-RIP140-MIN6) and the expression of green fluorescent protein (GFP) MIN6 cells (GFP-MIN6). Were given high fat and high glucose (25 rmmol/L glucose +500 mol/L palmitic acid) and / or 10/ mol/L. Pioglitazone intervention using MTT were detected in cells proliferation rate, the rate of apoptosis was detected by flow cytometry detection of RIP140 mRNA RT-PCR cells, Western blot detection of B lymphocyte in -2 (Bcl-2) expression were measured by thiobarbituric acid (MDA) and xanthine oxidase method to detect superoxide dismutase compounds (SOD) content. The results of N C缁，

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