目的探讨银杏内酯、银杏黄酮对蛛网膜下腔出血(SAH)及脑淋巴阻滞(CLB)后大鼠脑组织促凋亡基因Fas配体(FasL)表达的影响。方法将96只Wistar大鼠分为正常对照组、SAH组、SAH+CLB组、SAH+CLB+生理盐水组(NS组),以及银杏内酯高、低剂量组和银杏黄酮高、低剂量组。用结扎颈部淋巴管、摘除淋巴结及注入自体动脉血法制作CLB和SAH大鼠模型。制作CLB模型后1 d诱导SAH模型。用免疫荧光法和逆转录聚合酶链反应(RT-PCR)法检测大鼠脑组织FasL蛋白及mRNA的表达。结果各模型组大鼠脑组织FasL蛋白及mRNA表达明显高于正常对照组(P〈0.05~0.01),其中以SAH+CLB组、NS组表达最高。SAH组、银杏内酯和银杏黄酮高剂量组大鼠脑组织FasL蛋白及mRNA的表达水平明显低于SAH+CLB组、NS组(均P〈0.01),其中两高剂量组表达又明显低于低剂量组(均P〈0.01),两低剂量组大鼠脑组织FasL蛋白阳性细胞数与SAH+CLB组及NS组差异无统计学意义。结论 SAH+CLB能诱导脑组织FasL表达水平显著增高,银杏内酯和银杏黄酮对其有抑制作用,尤以高剂量组效果更明显。
Objective To investigate the effects of ginkgolide and ginkgetin on expression of pro-apoptotic Fas ligand(FasL) in cerebral tissue of subarachnoid hemorrhage(SAH) and cerebral lymphatic blockade(CLB) rats.Methods 96 Wistar rats were divided into normal control group,SAH group,SAH+CLB group,SAH+CLB+normal saline group(NS group),high and low dose ginkgolide groups,high and low dose ginkgetin groups.CLB model was made by ligation of cervical lymphatics and extirpation of lymphonode,and SAH model was made by injection of auto-arterial blood.SAH model was made 1 d after the establishment of CLB model.The expressions of FasL protein and mRNA in cerebral tissue were determined by immunohistochemistry and reverse transcription-polymerase chain reaction(RT-PCR) respectively.Results The expression of FasL protein and mRNA in cerebral tissue of the model rats were significantly higher than normal controls(P〈0.05~0.01).It was the highest in SAH + CLB group and NS group.The expression of FasL protein and mRNA in brain of SAH group and ginkgolide and ginkgetin high dose groups were significantly lower than SAH+CLB group and NS group(all P〈0.01).And the expression in the two high dose groups were significantly lower than in the two low dose groups(all P〈0.01).However,the FasL protein positive cells of the two low dose groups had no significant difference with SAH+CLB group and NS group.Conclusions SAH+CLB can increase the expression of FasL in cerebral tissue significantly.Ginkgolide and ginkgetin can inhibit it,especially in high dose group.