天麻素对脑缺血再灌注损伤大鼠神经修复及水通道蛋白4的影响

Effects of gastrodin on nerve repair and AQP4 after ischemic re-injection of brains in rats

目的:探讨天麻素(GAS)对大鼠脑缺血再灌注损伤(CIRI)后神经修复及水通道蛋白4(AQP4)的影响。方法:将健康雄性SD大鼠75只随机分为假手术组(sham)、大脑中动脉栓塞组(MCAO)、低剂量GAS组(GAS-L)、中剂量GAS组(GAS-M)及高剂量GAS组(GAS-H),每组为15只。将GAS溶解于生理盐水配制成不同剂量进行灌胃,sham组及大脑中动脉栓塞(MCAO)组用等剂量的生理盐水灌胃,1次/d,连续灌胃7 d,建立MCAO模型后每组按上述剂量继续灌胃7 d后测神经功能评分及体重,生化检测肝、肾功能;HE及尼氏染色观察缺血半暗带区病理形态学变化;免疫组化观察血管界膜胶质细胞纤维酸性蛋白(GFAP)的表达;免疫荧光观察AQP4蛋白的表达;luxol fast blue(LFB)观察神经纤维的变化;real time RT-PCR检测β-catenin、GSK-3βmRNA的表达。结果:GAS灌胃后对各组大鼠肝、肾功能无统计学差异;与sham组比较,MCAO组体重显著下降(P<0.05),神经功能评分显著降低(P<0.05),神经元损伤严重、缺血半暗带组织水肿、血管界膜及神经纤维严重破坏,AQP4的表达显著增多(P<0.01),β-catenin、GSK-3βmRNA的表达显著上升(P<0.05);与MCAO组相比,GAS-L组、GAS-M组及GAS-H组体重下降较缓慢且与浓度递增呈负相关(P<0.05),神经功能评分显著升高(P<0.05),缺血半暗带组织水肿减轻、神经元损伤下降,血管胶质界膜逐渐完整,神经纤维损伤逐渐修复,AQP4蛋白表达显著降低(P<0.01),β-catenin、GSK-3βmRNA的表达显著下降(P<0.05),且它们均呈量效关系。结论:GAS抑制Wnt/β-catenin信号通路与AQP4表达对大鼠脑缺血再灌注损伤发挥神经保护作用。

Objective:The effects of gastrodin(GAS)on nerve repair and AQP4 after ischemic re-injection damage(CIRI)in rats were discussed.Methods:Seventy-five healthy male SD rats were randomly divided into a pseudo-surgical group(sham),middle cerebral artery embolization group(MCAO),a low-dose GAS-L group(GAS-L),a medium-dose GAS-M group,and a high-dose GAS-H group,each with 15.Gas dissolved in physiological saline is formulated into GAS-L(15 mg/kg),GAS-M(30 mg/kg)and GAS-H(60 mg/kg)for gastric perfusion,arterial thrombosis in the sham group and brain The plug(MCAO)group used the same dose of physiological saline to fill the stomach,1 time/d,continuous gastric irrigation 7 d,after the establishment of the MCAO model,each group continued to fill the stomach at the above dose 7 d after nerve function score and weight,biochemical detection of liver and kidney function He and Nissl observed pathomorphic changes in ischemic semi-dark belt regions,immunohistilation observed the expression of the vascular membrane GFAP protein,immunofluorescence observed the expression of AQP4 protein,luxol fast blue(LFB)observed changes in nerve fibers,real-time RT-PCR detected the expression ofβ-catenin,GSK-3βmRNA.Results:There were no statistical differences in liver and kidney function(P<0.05)in rats after gas perfusion(P<0.05),compared with the sham group,the weight of the MCAO group decreased significantly(P<0.05),the neurological function score decreased significantly(P<0.05),and neuronal damage was severe ischemic semi-dark band tissue edema,severe damage to vascular membranes and nerve fibers,significant increase in expression of AQP4(P<0.01),significant increase in expression ofβ-catenin,GSK-3βmRNA(P<0.05)Compared with the MCAO group,the gas-L group,gas-M group and GAS-H group lost weight more slowly and were negatively correlated with the increase in concentration(P<0.05),and the neurofunction score increased significantly(P<0.05),ischemic semi-dark band tissue edema is reduced,neuronal damage is decreased,vascular glial membrane is gradually complete,nerve fiber damage is gradually repaired,AQP4 protein expression is significantly reduced(P<0.01),β-catenin,GSK-3βthe expression of mRNA decreased significantly(P<0.05),and they were all quantitatively effective.Conclusion:GAS inhibition of Wnt/β-catenin signaling pathways and AQP4 expression played a neuroprotective role in ischemic re-perfusion damage in rat brain.