NRAGE对肠上皮细胞凋亡的影响

发布时间：2018-07-04 17:08

本文选题：NRAGE + 缺血再灌注损伤　；参考：《四川大学学报(医学版)》2017年04期

【摘要】：目的探讨神经生长因子受体作用黑色素瘤抗原基因同源蛋白(neurotrophin receptor-interacting MAGE homolog,NRAGE)是否参与肠缺血再灌注(ischemia and reperfusion,I/R)损伤的过程及其对肠上皮细胞凋亡及occludin蛋白的影响。方法制备SD大鼠小肠I/R损伤模型,取肠组织行免疫组化染色,观察NRAGE蛋白的表达变化;体外肠上皮永生化细胞株IEC-6给予缺氧及复氧后观察细胞NRAGE的表达变化;将NRAGE过表达慢病毒(Lv-NRAGE组)、NRAGE干扰表达慢病毒(sh-NRAGE组)及对照慢病毒(Lv-control组)感染IEC-6细胞,并设置不加慢病毒的对照组(NC组),Western blot及RT-PCR观察各组细胞NRAGE蛋白及mRNA的表达变化,流式细胞术观察各组IEC-6细胞的凋亡情况,Western blot观察感染慢病毒后紧密连接蛋白occludin蛋白的表达变化。结果 I/R损伤后肠黏膜上皮细胞NRAGE表达较sham组升高(P0.01);体外IEC-6细胞缺氧6h后NRAGE蛋白的表达增加(P0.01),NRAGE mRNA表达升高(P0.01)。感染慢病毒48h后,相比于Lvcontrol组,Lv-NRAGE组早期凋亡率增加(P0.01),occludin蛋白表达降低(P0.01),sh-NRAGE组早期凋亡率降低(P0.01),occludin蛋白表达增高(P0.001)。结论 NRAGE参与了I/R损伤过程,并促进肠上皮细胞凋亡,降低occludin蛋白的表达。
[Abstract]:Objective to investigate the role of nerve growth factor receptor (NGF) receptor in the pathogenesis of neurotrophin receptor-interacting mage homologue gene (NRAGE) involved in the injury of (ischemia and reperfusionI / R and the effect of NGF receptor on apoptosis of intestinal epithelial cells and occludin protein. Methods the intestinal I / R injury model of SD rats was established, and the expression of NRAGE protein was observed by immunohistochemical staining in intestinal tissues of SD rats, and the changes of NRAGE expression in intestinal epithelial immortalized cell line IEC-6 were observed after hypoxia and reoxygenation in vitro. NRAGE overexpression lentivirus (Lv-NRAGE group) and control lentivirus (Lv-NRAGE group) were infected with IEC-6 cells. The expression of NRAGE protein and mRNA was observed by Western blot and RT-PCR in the control group (NC group). Apoptosis of IEC-6 cells in each group was observed by flow cytometry and the expression of tight junction protein (occludin) was observed by Western blot after infection with lentivirus. Results after I / R injury, the expression of NRAGE in intestinal mucosal epithelial cells was higher than that in sham group (P0.01), and the expression of NRAGE protein in IEC-6 cells was increased (P0.01) after hypoxia for 6 h (P0.01). 48 hours after infection with lentivirus, the early apoptosis rate of Lv-NRAGE group increased compared with that of Lvcontrol group (P0.01). The expression of cludin protein decreased (P0.01) in sh-NRAGE group (P0.01) and the expression of cludin protein increased (P0.001) in sh-NRAGE group. Conclusion NRAGE is involved in the process of I / R injury, promotes the apoptosis of intestinal epithelial cells and reduces the expression of occludin protein.
【作者单位】：重庆市第三军医大学西南医院普通外科;北京海军总医院普通外科;河南省信阳市明港镇897医院;
【基金】：国家自然科学基金面上项目(No.81370479)资助
【分类号】：R574

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