CAJ | 학술논문

目的了解缺血缺氧性脑损伤(HIBD)新生大鼠神经元胞浆Smac/Diablo、caspase-9的变化及其相互关系,同时观察参附注射液的干预效果.方法 HIBD模型制作参照Rice等方法.新生7dSD大鼠按照完全随机数字表法分为假手术组、生理盐水对照组与参附治疗组,再按照造模后不同观察时间点又分为3 h、6 h、12 h、24 h、3 d、7 d6个亚组,每亚组8只.参附治疗组于造模后即刻腹腔注射参附注射液10 mL/kg,1次/d,连续用药3 d.生理盐水对照组用生理盐水替代参附注射液,用法用量相同;假手术组仅切开颈正中皮肤和分离右侧颈总动脉,不结扎,不缺氧.Western blotting检测各时间点各组大鼠神经元胞浆中Smac/Diablo、活化caspase-9蛋白的表达.结果假手术组Smac/Diablo、caspase-9蛋白表达微弱.生理盐水对照组与参附治疗组2种蛋白造模后3 h即明显增加,与假手术组比较,差异有统计学意义(P＜0.05);24 h时2种蛋白表达达到峰值,3 d后明显下降,7 d时接近假手术组.其中参附治疗组Smac/Diablo蛋白表达量12 h、24 h、3 d、7 d时较生理盐水对照组明显下降,差异有统计学意义(P＜0.05);参附治疗组caspase-9蛋白12 h、24 h、3 d时较生理盐水对照组明显下降,差异有统计学意义(P＜0.05).结论 (1)HIBD新生大鼠脑皮层神经元存在Smac/Diablo蛋白胞浆转位,同时伴有胞浆活化caspase-9表达增加:参附可减少HIBD大鼠Smac/Diablo蛋白的胞浆转位,减少caspase-9的活化.(2)Smac/Diablo、caspase-9参与了HIBD新生大鼠迟发性神经元损伤的病理机制.
목적 료해결혈결양성뇌손상(HIBD)신생대서신경원포장Smac/Diablo、caspase-9적변화급기상호관계,동시관찰삼부주사액적간예효과.방법 HIBD모형제작삼조Rice등방법.신생7dSD대서안조완전수궤수자표법분위가수술조、생리염수대조조여삼부치료조,재안조조모후불동관찰시간점우분위3 h、6 h、12 h、24 h、3 d、7 d6개아조,매아조8지.삼부치료조우조모후즉각복강주사삼부주사액10 mL/kg,1차/d,련속용약3 d.생리염수대조조용생리염수체대삼부주사액,용법용량상동;가수술조부절개경정중피부화분리우측경총동맥,불결찰,불결양.Western blotting검측각시간점각조대서신경원포장중Smac/Diablo、활화caspase-9단백적표체.결과 가수술조Smac/Diablo、caspase-9단백표체미약.생리염수대조조여삼부치료조2충단백조모후3 h즉명현증가,여가수술조비교,차이유통계학의의(P＜0.05);24 h시2충단백표체체도봉치,3 d후명현하강,7 d시접근가수술조.기중삼부치료조Smac/Diablo단백표체량12 h、24 h、3 d、7 d시교생리염수대조조명현하강,차이유통계학의의(P＜0.05);삼부치료조caspase-9단백12 h、24 h、3 d시교생리염수대조조명현하강,차이유통계학의의(P＜0.05).결론 (1)HIBD신생대서뇌피층신경원존재Smac/Diablo단백포장전위,동시반유포장활화caspase-9표체증가:삼부가감소HIBD대서Smac/Diablo단백적포장전위,감소caspase-9적활화.(2)Smac/Diablo、caspase-9삼여료HIBD신생대서지발성신경원손상적병리궤제.
Objective To investigate the expressions of Smac/Diablo and caspase-9 and their corelations in the cytoplasm of cerebral cortex of neonatal rats with hypoxic-ischemic brain damage (HIBD), and the possible interference effects of Shenfu injection on them. Methods Postnatal 7-d SD rats were randomly divided into Shenfu treatment group (group SF), normal saline group (group C) and sham-operated group (group S). Rat models of HIBD were established according to Rice's method.Based on the observing times after the success of model making, each group (n=48) was equally divided into 6 subgroups (3, 6, 12 and 24 h, and 3 and 7 d observing groups). Rats in the group S were not performed ligation of the right common carotid artery and not underwent hypoxia but only performed the separation of right common carotid artery. Shenfu injection was administered by intraperitioneal injection right after H/I insult and then once daily at a dosage of 10 mL/kg for 3 d into the rats of group SF. Saline was also administered into the rats of group C by the same methods and at the same dosage. Cytoplasm proteins were extracted from the cerebral cortex tissue, and then, Western blotting was employed to detect the protein levels of Smac/Diablo and activated caspase-9 in cytoplasm. Results Western blotting showed that the expression levels of Smac/Diablo and activated caspase-9 in cytoplasm of group S were very low; the expression levels of Smac/Diablo and activated caspase-9 in cytoplasm of group SF and group C were increased markedly since the 3rd h of the success of model making as compared with those in group S (P＜0.05): these in the 2 groups reached their peak levels 24 h after H/I insult, gradually decreased 3 d after H/I insult, and almost returned to the original levels 7 d after H/I insult. The expressions of Smac/Diablo in cytoplasm of group SF 12, 24 h and 3 d after H/I insult was significantly lower than that of group C (P＜0.05); the expression of activated caspase-9 in cytoplasm of group SF 12,24 h and 3 d after H/I insult was significantly decreased as compared with that of group C (P＜0.05).Conclusion Smac/Diablo cytoplasm translocation and increased expression level of caspase-9 appear in the cytoplasm of cerebral cortex cells after H/I injury; Shenfu injection can inhibit the expressions of Smac/Diablo and caspase-9; Smac/Diablo and caspase-9 involve in the pathophysiological mechanisms of neuronal damage after H/I.