内吗啡肽对过氧化氢诱导的大鼠胰岛损伤的保护作用  

Effect of endomorphins on H2O2-induced islet injury in rats

作　　者：田利民[1] 高翠霞[2] 刘静[1] 

机构地区：[1]甘肃省人民医院内分泌科,兰州730000 [2]甘肃省人民医院超声诊断科,兰州730000

出　　处：《中华糖尿病杂志》2011年第2期156-160,共5页CHINESE JOURNAL OF DIABETES MELLITUS

摘　　要：目的 研究过氧化氢（H2O2）对胰岛的损伤及内吗啡肽（EMs）对这种损伤的保护作用.方法 成年雄性Wistar大鼠18只,体质量200～300 g,体外分离、纯化和培养Wistar大鼠胰岛,双硫腙（DyTZ）染色鉴定,锥虫蓝染色检测细胞活力,葡萄糖刺激实验检测胰岛生物活性.四唑蓝（MTT）分析不同浓度的H2O2对胰岛的损伤;放射免疫法检测EMs对H2O2诱导的胰岛细胞胰岛素分泌量的影响;流式细胞技术检测EMs对H2O2诱导的细胞周期影响.Western blot检测EMs和H2O2对p21蛋白表达的影响.多组间均数比较采用方差分析.结果 胰岛获得率为95%±2%,活力为96%±3%,生物学功能正常.与对照组比较,H2O2处理组胰岛细胞胰岛素释放量降低[分别为（67±9）mU/L,（153±11）mU/L,P＜0.01],而EMs＋H2O2组胰岛素释放量与H2O2组相比增加[分别为（108±13）mU/L,（116±12）mU/L,P＜0.05];H2O2处理组细胞周期G0/G1期细胞数目与对照组比较增多（分别为63.9%±3.1%,41.3%±1.9%,P＜0.05）;S期的细胞数目与对照组比较减少（分别为27.2%±1.1%、35.3%±2.6%,P＜0.05）;而用EM1预孵后与只加H2 O2处理的细胞比较,G0/G1期细胞数目减少（44.4%±2.1%,66.9%±3.1%,P＜0.05）,EM2预孵后再加H2O2处理的细胞与只加H2O2处理的细胞比较G0/G1期细胞数目减少（分别为47.6%±2.8%,66.9%±3.1%,P＜0.05）,EM1预孵后与只加H2O2处理的细胞比较S期的细胞数目增加（分别为32.1%±1.2%,24.2%±1.1%,P＜0.05）,EM2预孵后与只加H2O2处理的细胞比较S期的细胞数目增加（分别为31.2%±1.8%,24.2%±1.1%,P＜0.05）.H2O2处理组细胞p21蛋白表达增加（1.10±0.16,P＜0.05）,而EMs组细胞p21蛋白表达降低（分别为0.35±0.05,0.30±0.04,P＜0.05）.结论 内吗啡肽对过氧化氢诱导的胰岛损伤具有保护作用,其可能是通过降低p21蛋白表达和抗氧化损伤等机制实现的.Objective To evaluate the effects of endomorphin （EMs） on pancreatic islet injuries induced by H2O2. Methods Eighteen wistar rats＇ islets were isolated and purified,then identified by dithizone staining. The viability was determined by the Trypanblue exclusion assay. The function of the islets was examined by the insulin releasion response to glucose stimulation. MTT assay was used to analysis the effects of EMs on pancreatic islet injuries induced by H2O2. The medium insulin accumulation was measured by radioimmunoassay. The cell cycle distribution was analyzed by PI staining flow cytometric analysis. The protein level of p21 was detected by western blot. Results In the cultured cells in vitro,the ratio of islet cells was 95% ±2% and the cell viability was 96% ± 3%. Glucose stimulation experiments identified the islet cells could execute normal biology function. Compared with control group （ （ 153 ± 11 ） mU/L）,insulin accumulation of cells supernatant in H2O2 groups （ （ 67 ± 9 ） mU/L） was significantly decreased （ P 〈 0. 01 ）,while EMs enhanced the insulin accumulation of cells supernatant under H2O2 （ P 〈0. 05 ）. The cell number of cell cycle G0/G1 phase in H2O2 treatment group （63.9% ±3. 1% ） was significantly higher than that in control group（41.3% ± 1.9% ）,and S phase cell number in H2O2 treatment group （27.2% ± 1.1% ） was significantly lower than control （ 35. 3% ± 2.6% ）. The cell number of cell cycle G0/G1 phase in H2O2 treatment group induced by EM1 （44.4% ± 2. 1% ） was significantly lower than that in H2O2 treatment group （66.9% ±3. 1% ,P 〈0.05）,and S phase cell number in H2O2 treatment group induced by EM1 （ 32. 1% ± 1.2% ） was significantly higher than H2O2 group （24. 2% ± 1. 1%,P 〈 0. 05 ）. The results also showed the cell number of cell cycle Go/G1 phase in H2O2 treatment group induced by EM2 （47.6% ±2. 8% ） was significantly lower than that in H2O2 group（66. 9% ± 3. 1%,P 〈0. 05）,and S phase cell number in

关 键 词：内吗啡肽 胰岛 过氧化氢 氧化应激 

分 类 号：R587.1[医药卫生—内分泌]