CAJ | 학술논문

目的了解银杏内酯B(GKB)对癫痫大鼠海马中低氧诱导因子-1α(HIF-1α)与磷脂酰肌醇-3激酶/丝氨酸-苏氨酸蛋白激酶(PI3K/Akt)的影响,并探讨在这个过程中两者之间可能存在的关系.方法 21日龄SD大鼠96只随机分为5组:生理盐水组(NS组)、戊四氮(PTZ)致癫痫持续状态(SE)模型组(P组)、GKB治疗组(G+P组)、GKB治疗+PI3K抑制剂wortmannin干预组(G+P+W组)、wortmannin干预组(P+W组).于G+P组建模后1h、4h、8h、24 h,其他组建模后4h、8h处死大鼠取出脑组织,应用免疫组化方法及Western blot检测HIF-1α、p-Akt蛋白的表达.结果 (1)G+P组1h、4h、8h、24 h各时间点比较p-Akt蛋白表达差异有统计学意义(P＜0.01),4h表达达高峰(0.85±0.03);同时各时间点的HIF-1α蛋白表达差异有统计学意义(P＜0.01),8h表达达高峰(1.00±0.13).(2)8 h时间点各组间HIF-1α比较:NS组有少量表达,P组、G+P组表达量较NS组均明显升高(P＜0.01),且G+P组表达量高于P组(P＜0.01),G+P+W组、P+W组表达量明显降低,但G+P+W组高于P+W组(P＜0.01).(3)4h时间点各组间p-Akt比较:NS组有少量表达,P组、G+P组表达量较NS组均明显升高,且G+P组表达量高于P组(P＜0.01),G+P+W组、P+W组表达量均明显降低.结论在银杏内酯B作用下,癫痫持续状态大鼠海马内PI3K/Akt信号通路受到激活,并参与了海马内HIF-1α表达的调控.
목적 료해은행내지B(GKB)대전간대서해마중저양유도인자-1α(HIF-1α)여린지선기순-3격매/사안산-소안산단백격매(PI3K/Akt)적영향,병탐토재저개과정중량자지간가능존재적관계.방법 21일령SD대서96지수궤분위5조:생리염수조(NS조)、무사담(PTZ)치전간지속상태(SE)모형조(P조)、GKB치료조(G+P조)、GKB치료+PI3K억제제wortmannin간예조(G+P+W조)、wortmannin간예조(P+W조).우G+P조건모후1h、4h、8h、24 h,기타조건모후4h、8h처사대서취출뇌조직,응용면역조화방법급Western blot검측HIF-1α、p-Akt단백적표체.결과 (1)G+P조1h、4h、8h、24 h각시간점비교p-Akt단백표체차이유통계학의의(P＜0.01),4h표체체고봉(0.85±0.03);동시각시간점적HIF-1α단백표체차이유통계학의의(P＜0.01),8h표체체고봉(1.00±0.13).(2)8 h시간점각조간HIF-1α비교:NS조유소량표체,P조、G+P조표체량교NS조균명현승고(P＜0.01),차G+P조표체량고우P조(P＜0.01),G+P+W조、P+W조표체량명현강저,단G+P+W조고우P+W조(P＜0.01).(3)4h시간점각조간p-Akt비교:NS조유소량표체,P조、G+P조표체량교NS조균명현승고,차G+P조표체량고우P조(P＜0.01),G+P+W조、P+W조표체량균명현강저.결론 재은행내지B작용하,전간지속상태대서해마내PI3K/Akt신호통로수도격활,병삼여료해마내HIF-1α표체적조공.
Objective To explore the effects of Ginkgolide B on the expression of hypoxia inducible factor 1 α (HIF-1α) and P I3K/Akt pathway in the hippocampus of developing rats after pentylenetetrazol(PTZ)-induced status epilepticus,and to investigate the correlation between HIF-1α expression and PI3K/Akt pathway.Methods Ninety-six SD rats aged 21 days were randomly divided into normal saline group(group NS),status epilepticus group (group P),GKB treatment groups (group G+P),GKB +wortmannin treated group (group G+P+W),wortmannin treated group(group P+W).The brain tissue were harvested from the rats at 4 and 8 hours after the inducement,but in the group G+P at 1 h,4 h,8 h,24 h.Immunohistochemistry and Western blot were used respectively to detect HIF-1α and p-Akt protein expression.Results (1) For the group G+P,there were statistical differences in the expression levels of p-Akt protein between 1 h,4 h,8 h and 24 h(P＜0.01),The p-Akt protein reached the peak level at 4 hours (0.85±0.03),there were statistical differences in the expression levels of HIF-1α protein between 1 h,4 h,8 h and 24 h(P＜0.01),the HIF-1α expression reached the peak level at 8 hours(1.00±0.13).(2) The expression of HIF-1α in all the groups at 8 hours time point:the expression levels of HIF-1α in the group P and group G+P were significantly higher than those in the group NS (P＜0.01) and the expression levels of HIF1α in the group G+P were higher than those in the group P(P＜0.01).Using wortmannin,the PI3K/Akt specific inhibitor,HIF-1α protein expression in the group G+P+W and P+W was significantly decreased when compared with the group G+P and P (P＜0.01).(3)The expression of p-Akt in all the groups at 4 hours time point:the expression levels of p-Akt in the group P and group G+P were significantly higher than those in the group NS (P＜0.01) and the expression levels of p-Akt in the group G+P were higher than those in the group P (P＜ 0.01).Using wortmannin,p-Akt protein expression in the group G+P+W and P+W was significantly decreased when compared with the group G+P and P (P＜0.01).Conclusion GKB can activate PI3K/Akt signaling pathway,and the pathway is involved in regulating the expression of HIF-1α.