龙琥醒脑颗粒对脑出血模型小鼠cAMP/PKA/AQP4信号通路的调控作用

摘要

Objective:To investigate the effect of Longhu Xingnao granule (LXG) on intracerebral hemorrhage in rats and the regulation of cAMP/PKA/AQP4 signaling pathway.Methods:A mouse ICH model was established by Rynkowski method.Sixty male ICH mice were randomly divided into normal group,sham operation group,model group,LXG high-dose (20 g/kg) group and low-dose (10 g/kg) group.Mice in high-dose and low-dose groups were given LXG via gavage for two weeks after modeling,and mice in other groups were given equal volume of saline.Neurological deficits was assessed by Longa'method on day 7 and day 14 after administration.Brain edema was quantitated by dry/wet weight.The mRNA and protein expressions of cAMP,PKA and AQP4 in brain tissues were detected by qPCR and western blotting,respectively.Results:Neurological deficits grade and brain water content were markedly increased in the model group when compared with the normal group and sham operation group (P＜0.05).Compared with model group,neurological deficits grade and brain tissue water content were obviously decreased in LXG high-dose group and low-dose group (P＜0.05).The mRNA and protein expressions of cAMP,PKA and AQP4 in brain tissues were notably down-regulated in LXG high-dose group and low-dose group (P＜0.05).Conclusion:LXG attenuated neurological impairment and encephaledema in ICH mice probably via the regulation of the cAMP/PKA/AQP4 signaling pathway.%目的:探讨龙琥醒脑颗粒对脑出血(ICH)模型小鼠cAMP/PKA/AQP4信号通路的调控作用.方法:参照Rynkowski法建立小鼠ICH模型,将60只雄性ICH小鼠随机分为正常组、假手术组、模型组、龙琥醒脑颗粒高剂量组和龙琥醒脑颗粒低剂量组,每组12只,造模成功后,中药高、低剂量组小鼠分别灌胃给予20 g/kg、10 g/kg的龙琥醒脑颗粒溶液,正常对照组、假手术组和模型组小鼠则给予等量蒸馏水.采用Longa法,对各组小鼠分别于给药7d、14d后进行神经功能缺损评分;采用干湿重法评估ICH后血肿周围脑组织含水量;分别采用荧光定量PCR(qPCR)法和western blotting法检测末次给药后各组小鼠脑组织中cAMP、PKA和AQP4基因和蛋白表达.结果:与模型组比较,龙琥醒脑颗粒高、低剂量组均能显著降低神经功能缺损评分和脑组织含水量(P＜0.05),同时明显下调cAMP、PKA和AQP4基因和蛋白表达水平(P＜0.05).结论:龙琥醒脑颗粒对ICH模型小鼠cAMP/PKA/AQP4信号通路具有明显的调控作用,这可能是其抗神经功能缺损、减轻脑水肿而治疗ICH的重要机制之一.