CAJ | 학술논문

目的探讨发育期大鼠癫痫持续状态后海马内低氧诱导因子-1α(HIF-1α)表达与磷脂酰肌醇-3激酶(PI3K)/丝氨酸-苏氨酸蛋白激酶(Akt)信号通路之间的关系. 方法 21d龄SD大鼠54只按随机数字表法分为生理盐水组(n=24)、模型组(n=24)和渥曼青霉素干预组(n=6),其中模型组腹腔注射戊四氮(PTZ)诱导建立癫痫持续状态模型,生理盐水组腹腔注射等量生理盐水,渥曼青霉素干预组于癫痫持续状态模型建立前30 min腹腔注射0.5 mg/kg渥曼青霉素.分别于建模后1、4、8、24 h处死大鼠取出海马,应用免疫组化染色检测HIF-1α、Akt阳性细胞表达,Westernblottiong检测HIF-1α、Akt及磷酸化Akt(p-Akt)蛋白含量. 结果模型组海马内HIF-1α阳性细胞及蛋白于建模后1h即开始表达,4h明显升高,8h达高峰,24 h后下降;Akt阳性细胞及蛋白表达在各时间点间无明显变化;p-Akt蛋白于建模后1h即明显升高,4h达高峰,8h开始下降.生理盐水组各时间点HIF-1α阳性细胞及蛋白、p-Akt蛋白仅有极少量表达,其中模型组各时间点HIF-1α阳性细胞及蛋白表达均明显高于生理盐水组,差异有统计学意义(P＜0.05);Akt阳性细胞与蛋白表达与生理盐水组比较差异无统计学意义(P＞.05);建模后1、4、8h p-Akt蛋白表达明显高于生理盐水组,差异有统计学意义(P＜0.05).渥曼青霉素干预组HIF-1α、p-Akt蛋白表达于建模后4h有明显下降,与模型组比较差异均有统计学意义(P＜0.05). 结论发育期大鼠癫痫持续状态后可激活PI3K/Akt信号通路,并参与调控海马内HIF-1α的表达.
목적 탐토발육기대서전간지속상태후해마내저양유도인자-1α(HIF-1α)표체여린지선기순-3격매(PI3K)/사안산-소안산단백격매(Akt)신호통로지간적관계. 방법 21d령SD대서54지안수궤수자표법분위생리염수조(n=24)、모형조(n=24)화악만청매소간예조(n=6),기중모형조복강주사무사담(PTZ)유도건립전간지속상태모형,생리염수조복강주사등량생리염수,악만청매소간예조우전간지속상태모형건립전30 min복강주사0.5 mg/kg악만청매소.분별우건모후1、4、8、24 h처사대서취출해마,응용면역조화염색검측HIF-1α、Akt양성세포표체,Westernblottiong검측HIF-1α、Akt급린산화Akt(p-Akt)단백함량. 결과 모형조해마내HIF-1α양성세포급단백우건모후1h즉개시표체,4h명현승고,8h체고봉,24 h후하강;Akt양성세포급단백표체재각시간점간무명현변화;p-Akt단백우건모후1h즉명현승고,4h체고봉,8h개시하강.생리염수조각시간점HIF-1α양성세포급단백、p-Akt단백부유겁소량표체,기중모형조각시간점HIF-1α양성세포급단백표체균명현고우생리염수조,차이유통계학의의(P＜0.05);Akt양성세포여단백표체여생리염수조비교차이무통계학의의(P＞.05);건모후1、4、8h p-Akt단백표체명현고우생리염수조,차이유통계학의의(P＜0.05).악만청매소간예조HIF-1α、p-Akt단백표체우건모후4h유명현하강,여모형조비교차이균유통계학의의(P＜0.05). 결론 발육기대서전간지속상태후가격활PI3K/Akt신호통로,병삼여조공해마내HIF-1α적표체.
Objective To investigate the correlation between hypoxia-inducible factor-1α (HIF-1α) expression and activation of phosphatidyl inositol 3-kinase and serine/threonine kinase signal pathway in the hippocampus of developing rats after status epilepticus (SE).Methods Fifty-four SD rats aged 21 days were randomly divided into control group (n=24),SE group (n=24),wortmannin treatment group (n=6); SE rat models of the SE group were induced by intraperitoneal injection of 1％ 1,5-Pentamethylenetetrazole (PTZ); rats of the control group received injection of normal saline (NS); for wortmannin treatmnet group,the rats received intraperitoneal injection ofwortmannin 30 minutes before the inducement; the brain tissues were harvested from the rats at 1,4,8 and 24 h after the inducement,but only at 4 h in the wortmannin treatment group.The HIF-1α and Akt positive cells were detected with irnmunohistochemistry method.HIF-1α,Akt and p-Akt protein expressions were measured by Western blotting.Results In SE group,the HIF-1α expression began to occur at 1 h,significantly increased at 4 h after inducement,reached the peak level at 8 h,and began to decrease at 24 h; Akt protein positive cells showed no significant difference between each two time points; the p-Akt protein was significantly increased at 1 h,reached the peak level at 4 h and began to decrease at 8 h.However,the expression levels of HIF-1α and p-Akt protein in the control group were extremely low at each time point.So,the HIF-1α expression level in the SE vehicle group was significantly higher than that in the control group (P＜0.05); the p-Akt protein expression in SE group at 1,4 and 8 h was significantly higher than that in the control group (P＜0.05).The changes of Akt protein in the SE group were not time-dependent,and no significant difference was evident when it was compared with that of the control group (P＞0.05).Using wortmannin,the PI3K/Akt specific inhibitor,HIF-lα protein expression was significantly decreased when it was compared with the SE vehicle group (P＜0.05).Conclusion After status epilepticus in the developing rats,the PI3K/Akt signaling pathway is activated and the pathway involves in regulating the HIF-1α expression.