敲低Txnip抑制高糖诱导的人肾小管细胞系凋亡  

Txnip interference prevents HG-induced human kidney proximal tubular cell line apoptosis

作　　者：韦金英[1] 史永红[1] 任韫卓[1] 侯延娟[1] 杜春阳[1] 张连珊[1] 段惠军[1] 

机构地区：[1]河北医科大学病理学教研室,河北石家庄050017

出　　处：《基础医学与临床》2014年第1期16-21,共6页Basic and Clinical Medicine

基　　金：国家自然科学基金(81170673;81270804);高等学校博士学科点专项科研基金(20091323120001);河北省自然科学基金(C2011206147);河北省自然青年科学基金(H2012206132)

摘　　要：目的观察敲低Txnip对高糖诱导人肾小管上皮细胞凋亡的影响。方法将体外培养人肾小管上皮细胞分为正常糖组、高糖组、高糖+质粒载体对照组及高糖+shRNA组。采用原位末端转移酶标记技术(TUNEL)检测细胞凋亡;流式细胞术检测细胞ROS水平;Western blot检测caspase-3、cleaved caspase-3、BAX、BCL-2、P38 MAPK、P-P38 MAPK及细胞色素c的表达。结果与正常糖组相比,高糖组肾小管上皮细胞ROS产生和细胞凋亡明显增加(P<0.01),cleaved caspase-3和P-P38 MAPK表达增高,BAX/BCL-2比率明显升高以及细胞色素c易位(P<0.05)。敲低Txnip能够显著抑制高糖诱导的肾小管上皮细胞凋亡和ROS产生,下调cleaved caspase-3、和P-P38MAPK的表达,减少BAX/BCL-2比率和细胞色素c易位(P<0.05)。结论敲低Txnip能够抑制高糖诱导的人肾小管上皮细胞凋亡可能是通过减少ROS产生,保护线粒体功能,抑制P38MAPK信号通路激活而实现的。Objective To investigate the effect of Txnip interference on high glucose（ HG）-induced apoptosis in human kidney proximal tubular cell line（ HK-2）. Methods Cultured HK-2 were divided into normal glucose group（NG）,high glucose group（ HG）,HG ＋ contol plasmid vector（ HG ＋ C） and GH ＋ VDUP1 shRNA Plasmid（h）（HG ＋ shRNA）. Apoptosis of HK-2 was analyzed by DeadEndTMFluorometric TUNEL System. ROS produc-tion was observed by flow cytometry. The expression level of Txnip,caspase-3,cleaved caspase-3,BAX,BCL-2, P38 MAPK,P-P38 MAPK and cytochrome c protein was observed by Western blot. The expression level of Txnip, BAX,BCL-2 mRNA was observed by RT-PCR. Results Compared with normal glucose group（NG）,the production of ROS,the number of cell apoptosis,the expression of cleaved caspase-3 and P-P38 MAPK,ratio of BAX / BCL-2 and the release of cytochrome c from mitochondria to cytoplasm all significantly increased in HK-2 in high glucose group（HG）（P 〈0. 05）. Txnip interference inhibited HG-induced ROS production,cell apoptosis,expression of cleaved caspase-3 and P-P38 MAPK,ratio of BAX / BCL-2 and release of cytochrome c from mitochondria to cytopl-asm in HK-2（P 〈0. 05）. Conclusions Txnip interference can prevent HG-induced HK-2 apoptosis through decrea-sing ROS production,preserving mitochondrial function and inhibiting activation of P38 MAPK.

关 键 词：糖尿病肾病 氧化应激 人肾小管上皮细胞 凋亡 线粒体 

分 类 号：R587.2[医药卫生—内分泌]