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,P<0.01,48 h:t=24.00,P<0.01)。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,P<0.01)。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,P<0.01)及(31.38±3.92)%(vs高糖高脂组,t=5.20,P<0.01)。Bcl-2相对表达量分别为(1.14±0.06)、(0.42±0.02)(vs NC组,t=20.52,P<0.01)及(0.86±0.04)(vs高糖高脂组,t=17.71,P<0.01)。RIP140表达量分别为(1.13±0.11)、(2.34±0.21)(vs NC组,t=9.69,P<0.01)及(1.63±0.13)(vs高糖高脂组(t=5.03,P<0.01);高糖高脂组与NC组比较,MDA[(10.13±0.47vs(5.00±0.26)nmol/mg,t=16.57,P<0.01]、SOD[(5.15±1.07)协(12.25±1.25)nmol/mg,t=7.51,P<0.01]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,P<0.01]、SOD[(5.15±1.07)v5(8.74±0.59)nmol/mg,t=5.16,P<0.01)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,P<0.01),48 h(1.16±0.13)vs(1.
Objective To investigate the protective effect of Pioglitazone, a kind of peroxisome proliferators activated receptor （PPAR-Y）agonist, on the glucolipotoxicity-induced injury of 13 cells, and also evaluate the role of receptor interaction protein 140（RIP140） in this process. Methods MIN6 cells were divided into three groups： control group, high glucose and high fat group （Glucolipotoxicity group）, and Pioglitazone treated group （Pioglitazone group）. Overexpressed-RIP140-MIN6 （O-RIP140-MIN6） and overexpressed GFP-MIN6 （GFP-MIN6） were treated with high glucose and high fat （25 mmol/L glucose and 500 μmol/L palmitic acid） and/or 10/xmol/L Pioglitazone. The viability anct apoptosis of MIN6 cells was detected by MTT and flow cytometry respectively. RIP140 mRNA and Bcl-2 （B-cell lymphoma-2） level was analyzed by real-time PCR and western-blot respectively. Maleicdialdehyde （MDA） was tested by thiobarbituric acid method. Superoxide dismutase （SOD） was measured by xanthine oxidase method. Results MTT absorbance values were （1.80±0.04）、（0.95±0.04）and（0.97±0.03）;48 h（2.70±0.11）、（1.04±0.06）and（1.30±0.03） for 48 h in NC, Glucolipotoxicity and Pioglitazone group respectively. It is significantly different between NC group and Glucolipotoxicity group （24 h：t=25.94,P〈0.01,48 h：t=24.00,P〈0.01）. It is also significantly different between Glucolipotoxicity group and Pioglitazone group （48 h： t = 9.37, P〈0.01）. The 24 h apoptosis rates in each group was （2.93 ± 0. 66） %, （48.08± 3.95） M （vs NC group, t= 19.54, P〈0. 01） and（31.38± 3.92） % （vs Glucolipotoxicity group,t=5.20,P%0. 01） respectively. Bcl-2 relative expression level was （1.14±0. 06）, （0. 42±0.02） （vs NC group,t=20.52,P%0. 01）and（0. 864-0. 04） （vs Glucolipotoxicity group,t= 17. 71,P〈O. 01） respectively. RIP140 expression level was （1.13±0.11）, （2.34±0. 21）（vs NC group,t=9.69,P%0. 01） and （1.63±0. 13）[vs Glucoli
出处
《中国糖尿病杂志》
CAS
CSCD
北大核心
2016年第12期1094-1099,共6页
Chinese Journal of Diabetes
基金
国家自然科学基金(81170769)
长江大学联合项目基金(WJ2016-YZ-06)
