眼科研究
眼科研究
안과연구
CHINESE OPHTHALMIC RESEARCH
2010年
1期
45-49
,共5页
许惠卓%董淑倩%夏晓波%熊思齐
許惠卓%董淑倩%夏曉波%熊思齊
허혜탁%동숙천%하효파%웅사제
促红细胞生成素受体%人脐静脉内皮细胞%高糖
促紅細胞生成素受體%人臍靜脈內皮細胞%高糖
촉홍세포생성소수체%인제정맥내피세포%고당
erythropoietin receptor%human umbilical vein endothelial cell%high glucose
目的 探讨高糖条件下促红细胞生成素(EPO)受体mRNA及蛋白在人脐静脉内皮细胞(UVECs)的表达差异.方法 体外常规培养人UVECs细胞株,实验组分别给予22mmol/L葡萄糖作用12、24、48、72h,5.5mmol/L葡萄糖组(生理糖浓度)设为正常对照组,5.5mmol/L葡萄糖+16.5mmol/L甘露醇组为渗透压对照组.采用逆转录聚合酶链反应(RT-PCR)、免疫细胞化学法对高糖条件下人UVECs EPO受体mRNA和蛋白的表达进行检测.结果 正常对照组和渗透压对照组相比,人UVECs EPO受体mRNA和蛋白表达差异均无统计学意义(P>0.05).人UVECs在22mmol/L高糖作用12、24、48、72h后,EPO受体mRNA和蛋白表达均显著高于正常对照组(P<0.05),且随着时间的延长,EPO受体mRNA和蛋白的表达均逐渐增加.结论 高糖条件下人UVECs EPO受体mRNA和蛋白表达均增强,且呈时间依赖性.人UVECs在22mmol/L高糖条件下EPO受体mRNA及蛋白表达的增加与渗透压无关.
目的 探討高糖條件下促紅細胞生成素(EPO)受體mRNA及蛋白在人臍靜脈內皮細胞(UVECs)的錶達差異.方法 體外常規培養人UVECs細胞株,實驗組分彆給予22mmol/L葡萄糖作用12、24、48、72h,5.5mmol/L葡萄糖組(生理糖濃度)設為正常對照組,5.5mmol/L葡萄糖+16.5mmol/L甘露醇組為滲透壓對照組.採用逆轉錄聚閤酶鏈反應(RT-PCR)、免疫細胞化學法對高糖條件下人UVECs EPO受體mRNA和蛋白的錶達進行檢測.結果 正常對照組和滲透壓對照組相比,人UVECs EPO受體mRNA和蛋白錶達差異均無統計學意義(P>0.05).人UVECs在22mmol/L高糖作用12、24、48、72h後,EPO受體mRNA和蛋白錶達均顯著高于正常對照組(P<0.05),且隨著時間的延長,EPO受體mRNA和蛋白的錶達均逐漸增加.結論 高糖條件下人UVECs EPO受體mRNA和蛋白錶達均增彊,且呈時間依賴性.人UVECs在22mmol/L高糖條件下EPO受體mRNA及蛋白錶達的增加與滲透壓無關.
목적 탐토고당조건하촉홍세포생성소(EPO)수체mRNA급단백재인제정맥내피세포(UVECs)적표체차이.방법 체외상규배양인UVECs세포주,실험조분별급여22mmol/L포도당작용12、24、48、72h,5.5mmol/L포도당조(생리당농도)설위정상대조조,5.5mmol/L포도당+16.5mmol/L감로순조위삼투압대조조.채용역전록취합매련반응(RT-PCR)、면역세포화학법대고당조건하인UVECs EPO수체mRNA화단백적표체진행검측.결과 정상대조조화삼투압대조조상비,인UVECs EPO수체mRNA화단백표체차이균무통계학의의(P>0.05).인UVECs재22mmol/L고당작용12、24、48、72h후,EPO수체mRNA화단백표체균현저고우정상대조조(P<0.05),차수착시간적연장,EPO수체mRNA화단백적표체균축점증가.결론 고당조건하인UVECs EPO수체mRNA화단백표체균증강,차정시간의뢰성.인UVECs재22mmol/L고당조건하EPO수체mRNA급단백표체적증가여삼투압무관.
Objective Although vascular endothelial growth factor(VEGF)is a primary mediaor in diabetic retinopathy(DR).VEGF inhibition alone is insufficient for preventing retinal neovascularization.Some studies showed that erythropoietin(EPO)is a potent retinal angiogenic factor of independent of VEGF in DR.The present study is to investigate the effect of high glucose on the expression of mRNA and protein of the erythropoietin receptor(EPOR)in cultured human umbilical vein endothelial cells(UVECs)in vitro.MethodsHuman UVECs from the cell center of the hospital were cultured in vitro and passaged in DMEM containing 10% neonatal bovine serum with 22mmol/L of glucose for 12,24,48 and 72 hours in the experimental group.Cells cultured in 5.5mmol/L glucose were used as control group Ⅰ and mannitol + 22mmol/L of glucose(isotope)as control group Ⅱ.The expression of EPOR mRNA in Human UVECs were detected by semi-quantitative reverse transcriptase(RT)-polymerase chain reaction(PCR)and detected at A_(260mm)/A_(280mm).The PCR product was calculated as the A value of EPOR mRNA amplification/the A value of GAPDH mRNA amplification.The expression of EPOR protein in Human UVECs was detected by immunocytochemistry.ResultsThe A_(260mm)/A_(280mm) value of EPOR mRNA receptor expression was 0.32±0.02 in the 5.5mmol/L glucose group,and 0.34±0.02 in the mannitol+22mmol/L glucose group(P>0.05).In 12 hours,24 hours and 72 hours after the experiment,the A260mm/A280mm value of EPOR mRNA was 0.82±0.01,0.96±0.02 and 1.02±0.01,respectively,indicating a significant increase in comparison with the 5.5mmol/L glucose group.The expression of Human UVECs protein was gradually increased with passage in the experimental group.Expression of Human UVECs protein was stronger in various time points in the 22mmol/L glucose group than in the 5.5mmol/L glucose group.ConclusionHigh glucose elevates the expression of EPOR mRNA and protein in Human UVECs in a time-dependent manner.The effect of high glucose(22mmol/L glucose)on the expression of EPOR mRNA and protein in Human UVECs is not related to osmotic pressure.
