TXNIP过表达对INS-1细胞凋亡通路的影响  

Effect of adenovirus-mediated TXNIP overexpression on INS-I cell apoptosis pathway

作　　者：吴向征 张倩[1] 梁男男 刘晓波[1] 杨艳宏[1] 焦向英[1] 

机构地区：[1]山西医科大学生理学系,山西省太原市030001

出　　处：《中国心血管病研究》2016年第2期185-190,共6页Chinese Journal of Cardiovascular Research

基　　金：山西医科大学青年基金资助项目（项目编号：Q02201208）

摘　　要：目的本研究使用腺病毒转染技术，观察在正常糖脂浓度下培养的INS-1细胞过表达TXNIP是否引起细胞凋亡。并对TXNIP介导细胞凋亡的通路进行初步分析。方法将处于对数生长期在正常糖脂浓度下培养的INS-1细胞分为3组，即正常培养组、空病毒（Ad-eGFP）组和TXNIP过表达（Ad-TXNIP-eGFP）组，均于转染48h后收集细胞进行指标测定。结果转染细胞48h时病毒转染率达到高峰，并且荧光蛋白表达量基本一致。同Ad-eGFP组相比，Ad-TXNIP-eGFP组TXNIP mRNA（38．68±7．35比0．73±0．39，P〈0．01）和蛋白表达量（1．28±0．25比0．62±0．16，P〈0．01）均明显增高，说明转染及TXNIP过表达成功。与Ad-eGFP组相比，Ad-TXNIP-eGFP组Caspase-3相对活性明显增高[（3．823±0．238）nmol·h^-1·mg^-1比（0．956±0．107）nmol·h^-1·mg^-1。P〈0．01]；反映下游不同通路的Caspase-8[（132．10±27．33）pg／ml比（81．01±15．34）pg／ml，P〈0．01]、Caspase-9[（290．76±43．15）pg／ml比（s8．94±14．68）pg／ml，P〈0．01]和Caspase-12活性[（266．96±18．50）pg／ml比（52．05±6．13）pg／ml，P〈0．01]均明显增高。结论单纯过表达TXNIP可以引起正常糖脂浓度培养下INS-1细胞凋亡。线粒体凋亡途径、死亡受体活化途径和内质网应激介导途径均参与了TXNIP引起的INS-1细胞凋亡的发生。Objective To observe if TXNIP overexpression alone can cause INS-1 cell apoptosis, and the effect of TXNIP on different apoptotie pathways, adenovirus carrying TXNIP gene was used to transfect INS-1 islet cells cultured in normal glucose and normal lipid concentration conditions. Methods INS-1 islet cells in logarithmic growth phase were randomly divided into three groups： normal cultured group, empty adenovirus vector （Ad-eGFP）group and TXNIP overexpression （Ad-TXNIP-eGFP）group. All the cells were collected 48 h after transfection for further assay. Results Transfection efficiency reached its peak 48 h after the adenovirus transfection, and the expression of green fluorescent protein in three groups were nearly equally. Compared to the Ad-eGFP group, the expression of TXNIP mRNA（38.68±7.35 vs 0.73±0.39, P〈0.01 ） and protein （1.28±0.25 vs 0.62±0.16, P〈0.01 ） of Ad-TXNIP-eGFP group increased significantly, which meant adenovirus-mediated TXNIP overexpression was successful. Caspase-3 activity was significantly increased compared to Ad-eGFP group[ （3.823± 0.238 ）nmol · h^-1·mg^-1 vs （0.956 ±0.107 ）nmol · h^-1·mg^-1, P〈0.01 ]. To further elucidate the downward pathways may involve in TXNIP induced cell apoptosis, our results shown that Caspase-8± （132.10±27.33）pg/ml vs （81.01± 15.34）pg/ml, P〈0.01 ], Caspase-9 [（290.76±43.15）pg/ml vs （88.94±14.68）pg/ml, P〈0.01 ] and Caspase-12 activity [（266.96±18.50）pg/ml vs （52.05±6.13）pg/ml, P〈0.01 ± were all increased significantly when compared with the eGFP group. Conclusion TXNIP overexpression alone can cause INS-1 cell apoptosis, which were cultured in normal glucose and normal lipid concentration conditions. Furthermore, the apoptosis is mediated by three different pathways, including mitochondria apoptosis pathway, death receptor pathway and endoplasmic reticulum stress pathway.

关 键 词：硫氧还蛋白相互作用蛋白 INS-1细胞 内质网应激途径 死亡受体途径 线粒体 凋亡途径 凋亡 

分 类 号：R587.1[医药卫生—内分泌]