Necrostatin-1对TNF-α诱导的胰岛细胞损伤的保护作用  

Protective effect of necrostatin-1 on the damage of pancreas islet cells induced by TNF-α

作　　者：叶斌[1] 容鹏飞[1] 刘亮[2] 王维[1] 张声旺[1] YE Bin;RONG Pengfei;LIU Liang;WANGWei;ZHANG Shengwang(Department of Radiology,Third Xiangya Hospital,Central South University,Changsha 410013;Department of Radiology,Yiyang Central Hospital,Yiyang Hunan 413000,China)

机构地区：[1]中南大学湘雅三医院放射科,长沙410013 [2]益阳市中心医院放射科,湖南益阳413000

出　　处：《中南大学学报（医学版）》2020年第7期752-758,共7页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81903199)。

摘　　要：目的:研究necrostatin-1(Nec-1)是否对TNF-α介导的胰岛细胞损伤具有保护作用。方法:新生猪胰岛细胞分离纯化后分为3组:Nec-1组(在培养基中加入Nec-1+TNF-α)、TNF-α组(在培养基中加入TNF-α),以及纯培养基的对照组,每组均含胰岛10000 IEQ,培养48 h后观察细胞数量的变化;通过吖啶橙/溴乙锭(AO/EB)染色观察细胞的死亡状况;采用化学发光法检测胰岛素分泌量和核酸定量分析法检测胰岛细胞DNA含量;采用RT-PCR法检测胰岛素、胰高血糖素及生长抑素的mRNA表达水平;采用流式细胞术检测B细胞的活性。结果:Nec-1组、TNF-α组与对照组培养48 h后胰岛数量分别为(8425±2187),(4325±778)与(7122±1558)IEQ;AO/EB染色显示TNF-α组死亡的胰岛细胞较另外两组明显增多;Nec-1组、TNF-α组与对照组胰岛素/胰岛细胞DNA值分别为(13.21±3.15),(2.47±0.45)与(7.44±0.97)mIU/mg;与TNF-α组、对照组比较,Nec-1组胰岛素(6.73±1.07)、胰高血糖素(10.13±1.98)与生长抑素(8.57±1.11)基因的mRNA相对表达水平最高均(均P<0.05);Nec-1组活细胞比例(97.32±1.87)%、B细胞比例(90.86±3.68)%均较TNF-α组、对照组明显升高(均P<0.05)。结论:TNF-α可以诱导新生猪胰岛细胞的损伤,而Nec-1对TNF-α诱导的胰岛细胞损伤具有保护作用,Nec-1可能会提高新生猪胰岛细胞团中内分泌细胞的含量。Objective:To investigate whether necrostatin-1(Nec-1)can protect islet cells from the damage induced by TNF-α.Methods:After isolation and purification,the neonatal porcine islet cell clusters(NICCs)were divided into 3 groups(islets 10000 IEQ/group):a Nec-1 group(Nec-1+TNF-αwas added to the culture medium),a TNF-αgroup(TNF-αwas added to the culture medium),and a control group(pure medium).The number of cells was observed after 48 h of coculture.The cell death was evaluated by AO/EB staining.Insulin secretion and DNA of islets were detected by chemiluminescence and nucleic acid quantitative analysis.RT-PCR assay was used to examine the m RNA expressions of insulin gene,glueogan gene and somatostatin gene.Flow cytometry analysis was used to detect the viability of B cells.Results:The number of islets in Nec-1 group,TNF-αgroup and the control group were(8425±2187),(4325±778),and(7122±1558)IEQ,respectively.Compared to the other two groups,the number of dead cells in TNF-αgroup was greatly increased.The insulin/DNA values in the Nec-1 group,TNF-αgroup and blank control group were(13.21±3.15),(2.47±0.45),and(7.44±0.97)m IU/mg,respectively.Compared to the TNF-αgroup and the control group,the m RNA relative expression levels of insulin gene(6.73±1.07),glucagon gene(10.13±1.98),somatostatin gene(8.57±1.11)were significantly increased in the Nec-1 group(all P<0.05),the rate of live cells(97.32±1.87)%and live B cells(90.86±3.68)%were increased significantly in the Nec-1 group(all P<0.05).Conclusion:TNF-αcan induce neonatal porcine islet cells damage,which is attenuated in the presence of Nec-1.Nec-1 can increase the content of endocrine cells in NICCs.

关 键 词：糖尿病 胰岛 necrostatin-1 TNF-Α 

分 类 号：R587.1[医药卫生—内分泌]