CAJ | 학술논문

目的：在大鼠局灶性脑缺血模型中观察低氧反应元件（HRE）介导的神经营养因子-3（NT-3）缺血／低氧调控表达及其对缺血再灌注脑损伤的保护作用。方法将重组反转录病毒（RV-5HRE-NT3及RV-5HRE-EGFP）或0.9％氯化钠溶液（对照组）注射入大鼠脑内3 d后，用大脑中动脉线栓阻塞法（ tMCAO）制作大鼠局灶性脑缺血再灌注损伤模型。用免疫组织化学染色法和Western blot法检测NT-3的表达，用TTC染色法检测脑梗死体积，用TUNEL染色法检测细胞凋亡，用Reglodi评分法检测大鼠神经功能情况。结果大鼠tMCAO 3 d后RV-5HRE-NT3组中NT-3阳性细胞数明显高于对照组和RV-5HRE-EGFP组，且着色较深；tMCAO后第1、3、7和14天，RV-5HRE-NT3组中的NT-3蛋白表达均明显高于对照组和RV-5HRE-EGFP组（P＜0.05）。在RV-5HRE-NT3组中，tMCAO 3 d大鼠脑梗死体积百分比明显减小（ P＜0.05）， tMCAO 3和7 d后缺血半暗带凋亡细胞百分比也明显减少（ P＜0.05）。 RV-5HRE-NT3注射可以改善大鼠tMCAO后的神经功能障碍。结论 HRE介导的NT-3低氧反应性表达上调可以减轻大鼠局灶性脑缺血再灌注脑损伤。
목적：재대서국조성뇌결혈모형중관찰저양반응원건（HRE）개도적신경영양인자-3（NT-3）결혈／저양조공표체급기대결혈재관주뇌손상적보호작용。방법장중조반전록병독（RV-5HRE-NT3급RV-5HRE-EGFP）혹0.9％록화납용액（대조조）주사입대서뇌내3 d후，용대뇌중동맥선전조새법（ tMCAO）제작대서국조성뇌결혈재관주손상모형。용면역조직화학염색법화Western blot법검측NT-3적표체，용TTC염색법검측뇌경사체적，용TUNEL염색법검측세포조망，용Reglodi평분법검측대서신경공능정황。결과대서tMCAO 3 d후RV-5HRE-NT3조중NT-3양성세포수명현고우대조조화RV-5HRE-EGFP조，차착색교심；tMCAO후제1、3、7화14천，RV-5HRE-NT3조중적NT-3단백표체균명현고우대조조화RV-5HRE-EGFP조（P＜0.05）。재RV-5HRE-NT3조중，tMCAO 3 d대서뇌경사체적백분비명현감소（ P＜0.05）， tMCAO 3화7 d후결혈반암대조망세포백분비야명현감소（ P＜0.05）。 RV-5HRE-NT3주사가이개선대서tMCAO후적신경공능장애。결론 HRE개도적NT-3저양반응성표체상조가이감경대서국조성뇌결혈재관주뇌손상。
Objective To investigate the neuroprotective effects of neurotrophin-3 (NT-3) expression controlled by five copies of the hypoxia-responsive elements after focal cerebral ischemia .Methods Three groups of rats re-ceived RV-5H-NT3, RV-5H-EGFP or saline injection .Three days after gene transfer , the rats underwent 90 min of transient middle cerebral artery occlusion ( tMCAO) , followed by 1-28 days of reperfusion .Immunohistostaining and western blotting were performed to detect ischemia/hypoxia-regulated expression of NT-3 controlled by HRE . The volume of brain infarction and the apoptosis were analysised by TTC and TUNEL staining .The neurological scoring was determined by neurological behavior tests .Results Three days after tMCAO , brain NT-3 expression was significantly increased in the RV-5HNT3-transduced animals compared with the RV-5H-EGFP or saline group (P<0.05), and brain infarct volume was smaller in the RV-5H-NT3-transduced group than the RV-5H-EGFP or saline group ( P<0.05 ) .The percentage of TUNEL-positive cells was reduced in RV-5 H-NT3-transduced brains compared with the RV-5 HEGFP or saline group 3 and 7 days after tMCAO ( P<0.05 ) .Furthermore , the neurolog-ical status of RV-5H-NT3-transduced rats was better than that of RV-5H-EGFP-or saline-transduced animals from 1 day to 4 weeks after tMCAO ( P<0.05 ) .Conclusions HRE may modulate NT-3 expression in the ischemic brain tissue and that the up-regulated NT-3 may effectively improve neurological status following tMCAO due to de-creased initial damage .