3,4-二羟基苯甲酸乙酯预处理对白蛋白诱导低氧培养肾小管上皮细胞凋亡的影响  

Effect of ethyl-3,4 dihydroxybenzoate pretreatment on the tubular epithelial cells apoptosis induced by albumin and hypoxia

作　　者：王沛[1] 刘章锁[1] 梁献慧[1] 马瑨[1] 骆红[1] 石新慧[1] 

机构地区：[1]郑州大学第一附属医院肾内科河南省高等学校重点学科临床开放实验室,450052

出　　处：《中华肾脏病杂志》2010年第8期624-628,共5页Chinese Journal of Nephrology

摘　　要：目的 探讨脯胺酰羟化酶抑制剂3,4-二羟基苯甲酸乙酯（EDHB）对白蛋白诱导的低氧条件下培养的肾小管上皮细胞（NRK-52E）凋亡的影响.方法 NRK-52E细胞在以下各组孵育24 h：常氧（5%CO2＋空气）组,低氧（1%O2＋5%CO2＋94%N2）组,常氧＋白蛋门（30 g/L）组及低氧＋白蛋白（30 g儿）组,观察各组NRK-52E细胞的凋亡情况.然后NRK-52E细胞在以下各组孵育24 h：常氧组、低氧组,低氧＋白蛋白（30 g/L）组,低氧＋EDHB（500 μmol/L）组,EDHB预处理组（培养细胞中先加入EDHB 500 μmol/L,0.5 h后再加入BSA 30 g/L,低氧培养）,观察EDHB对白蛋白诱导低氧培养NRK-52E凋亡的影响.流式细胞技术检测细胞凋亡;RT-PCR检测凋亡相关蛋门bcl-2、bax及血管内皮生长因子（VEGF）mRNA表达;Western印迹检测VEGF蛋白表达.结果 NRK-52E细胞凋亡率在常氧组与低氧组差异无统计学意义（P〉0.05）,但低氧＋白蛋白组细胞凋亡率显著高于常氧＋白蛋白组（37.36%±4.95%比25.59%±3.32%,P〈0.05）.低氧＋白蛋白组bax mRNA表达显著高于常氧＋白蛋白组（P〈0.05）,而bcl-2 mRNA的表达则显著低于常氧＋白蛋白组（P〈0.05）.EDHB预处理可显著抑制低氧＋白蛋白组细胞凋亡率的增高（P〈0.05）、bax mRNA表达的增高（P〈0.05）以及bcl-2 mRNA表达的降低（P〈0.05）.低氧组NRK-52E细胞VEGF mRNA和蛋白表达显著高于常氧组（P〈0.05）,而低氧＋白蛋白组则显著低于低氧组（P〈0.05）.EDHB预处理可显著抑制低氧＋白蛋白组细胞VEGF mRNA和蛋白表达的降低（P〈0.05）.结论 白蛋白和低氧联合刺激可显著增加NRK-52E细胞凋亡,EDHB预处理可改善该病变,可能与其提高VEGF的表达有关.Objective To explore the effects of ethyl-3,4 dihydroxybenzoate（EDHB）, a prolyl hydroxylase inhibitor, pretreatment on the tubular epithelial cells apoptosis induced by albumin and hypoxia. Methods To investigate the effects of albumin and hypoxia on cells, rat tubular epithelial cells（NRK-52E）were incubated for 24 h in：（1）normoxia（5%CO2）;（2）hypoxia（1% O2）;（3）albumin（30 g/L）under normoxia;（4）albumin（30 g/L）under hypoxia. To investigate the effects of EDHB pretreatment on cells apoptosis, NRK-52E were incubated in hypoxia for 24 h in：（1）normoxia;（2）hypoxia;（3）hypoxia＋albumin（30 g/L）;（4）hypoxia＋EDHB（500 μmol/L）;（5）EDHB pretreatment（albumin 30 min after EDHB）. Apoptosis was measured by flow cytometry（AnnexinV-FITC-PI）. bcl-2, bax and vascular epithelial growth factor（VEGF）mRNA expression were detected by RT-PCR. VEGF protein expression was detected by Western blotting. Results NRK-52E apoptosis was not significantly different between hypoxia and norraoxia groups（P〉0.05）, but increased significantly in albumin（30 g/L）under hypoxia group compared with albumin（30 g/L）under normoxia group（37.36%？.95% vs 25.59%？.32%, P〈 0.05）. There was an increase in bax mRNA expression and a decrease in bcl-2 mRNA expression in albumin（30 g/L）under hypoxia group compared with albumin（30 g/L）under normoxia group（P〈 0.05）. EDHB pretreatment improved these impairments of albumin（30 g/L）under hypoxia on NRK-52E（P〈 0.05）. VEGF expression elevated in hypoxia compared with normoxia（P〈0.05）, decreased in albumin（30 g/L）under hypoxia groups compared with that without albumin groups（P〈0.05）.EDHB pretreatment significantly improved VEGF expression compared with albumin（30 g/L）under hypoxia group（P 〈0.05）. Conclusion NRK-52E cells apoptosis induced by albumin is accelerated by hypoxia, however partially improved by EDHB pretreatment, probably through the up-regulation of VEGF expression.

关 键 词：缺氧 自蛋白类 细胞凋亡 肾小管上皮细胞 脯胺酰羟化酶抑制剂 血管内皮生长因子 

分 类 号：R692[医药卫生—泌尿科学]