CAJ | 학술논문

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.
목적 탐토포알선간화매억제제3,4-이간기분갑산을지(EDHB)대백단백유도적저양조건하배양적신소관상피세포(NRK-52E)조망적영향.방법 NRK-52E세포재이하각조부육24 h:상양(5%CO2+공기)조,저양(1%O2+5%CO2+94%N2)조,상양+백단문(30 g/L)조급저양+백단백(30 g인)조,관찰각조NRK-52E세포적조망정황.연후NRK-52E세포재이하각조부육24 h:상양조、저양조,저양+백단백(30 g/L)조,저양+EDHB(500 μmol/L)조,EDHB예처리조(배양세포중선가입EDHB 500 μmol/L,0.5 h후재가입BSA 30 g/L,저양배양),관찰EDHB대백단백유도저양배양NRK-52E조망적영향.류식세포기술검측세포조망;RT-PCR검측조망상관단문bcl-2、bax급혈관내피생장인자(VEGF)mRNA표체;Western인적검측VEGF단백표체.결과 NRK-52E세포조망솔재상양조여저양조차이무통계학의의(P>0.05),단저양+백단백조세포조망솔현저고우상양+백단백조(37.36%±4.95%비25.59%±3.32%,P<0.05).저양+백단백조bax mRNA표체현저고우상양+백단백조(P<0.05),이bcl-2 mRNA적표체칙현저저우상양+백단백조(P<0.05).EDHB예처리가현저억제저양+백단백조세포조망솔적증고(P<0.05)、bax mRNA표체적증고(P<0.05)이급bcl-2 mRNA표체적강저(P<0.05).저양조NRK-52E세포VEGF mRNA화단백표체현저고우상양조(P<0.05),이저양+백단백조칙현저저우저양조(P<0.05).EDHB예처리가현저억제저양+백단백조세포VEGF mRNA화단백표체적강저(P<0.05).결론 백단백화저양연합자격가현저증가NRK-52E세포조망,EDHB예처리가개선해병변,가능여기제고VEGF적표체유관.
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.