穿心莲内酯对脑出血大鼠脑水肿及AQP4、NKCC1表达的影响  

Effect of Andrographolide on Cerebral Edema and the Expression of AQP4 and NKCC1 in Rats with Cerebral Hemorrhage

作　　者：姜燕强 陈俊丽 陈利平 段国华 张昆 刘亚利 刘英 JIANG Yanqiang;CHEN Junli;CHEN Liping;DUAN Guohua;ZHANG Kun;LIU Yali;LIU Ying(Wu′an First People′s Hospital,Handan 056300,Hebei,China)

机构地区：[1]武安市第一人民医院,河北邯郸056300

出　　处：《中西医结合心脑血管病杂志》2022年第11期1974-1980,共7页Chinese Journal of Integrative Medicine on Cardio-Cerebrovascular Disease

基　　金：河北省医学科学研究课题(No.20201590)。

摘　　要：目的探讨穿心莲内酯(AND)对脑出血大鼠脑水肿及水通道蛋白4(AQP4)、钠-钾-氯协同转运蛋白1(NKCC1)的影响及其机制。方法采用随机数字表法将160只SD大鼠分为假手术组、模型组、穿心莲内酯50 mg/kg组和尼莫地平10 mg/kg组,每组40只。采用自体血注射法制备脑出血大鼠模型,造模完成后,尼莫地平10 mg/kg组给予2 mg/mL尼莫地平溶液(灌胃体积5 mL/kg)灌胃,每日1次;穿心莲内酯50 mg/kg组给予10 mg/mL穿心莲内酯溶液(灌胃体积5 mL/kg)灌胃,每日1次;假手术组和模型组给予生理盐水灌胃5 mL/kg,每日1次。各组疗程7 d。采用改良神经功能缺损评分标准(mNSS)进行神经功能缺失评分,通过多田公式计算脑血肿体积,失重法检测脑含水量,伊文思蓝(EB)法测定血脑屏障通透性;通过透射电子显微镜(TEM)观察血脑屏障超微结构变化;免疫荧光法和蛋白免疫印迹(Western Blot)法检测脑组织AQP4和NKCC1表达。结果模型组大鼠神经功能缺失评分、脑血肿体积、脑含水量、血脑屏障通透性均明显高于假手术组(P<0.01);穿心莲内酯50 mg/kg组和尼莫地平10 mg/kg组神经功能缺失评分、脑血肿体积、血脑屏障通透性均明显低于模型组(P<0.01);且穿心莲内酯50 mg/kg组神经功能缺失评分、脑血肿体积、脑含水量、血脑屏障通透性均明显低于尼莫地平10 mg/kg组(P<0.05或P<0.01)。通过TEM观察发现,模型组大鼠血脑屏障可见基底膜薄厚不均匀、膨大、断裂现象,紧密连接结构(TJPs)疏松,内皮细胞皱缩等超微结构病变;与模型组比较,穿心莲内酯50 mg/kg组和尼莫地平10 mg/kg组血脑屏障超微结构病变不同程度减轻,其中穿心莲内酯50 mg/kg组效果更为明显。通过免疫荧光法和Western Blot法检测发现,模型组大鼠脑组织AQP4、NKCC1蛋白表达量明显高于假手术组(P<0.01);穿心莲内酯50 mg/kg组和尼莫地平10 mg/kg组AQP4、NKCC1蛋白表达量明显低于模型组(P<0.01);且Objective To investigate the effects of andrographolide(AND)on cerebral edema and the expression of aquaporin 4(AQP4)and sodium potassium chloride cotransporter 1(NKCC1)in rats with cerebral hemorrhage and its mechanism.Methods One hundred and sixty Sprague-Dawley(SD)rats were divided into sham operation group,model group,AND50 mg/kg group,and nimodipine 10 mg/kg group,with 40 rats in each group by random number table method.The intracerebral hemorrhage rat model was prepared by autologous blood injection.After modeling,nimodipine 10 mg/kg group was given 2 mg/mL nimodipine solution(gavage volume 5 mL/kg),once a day.AND50 mg/kg group was given 10 mg/mL andrographolide solution(gavage volume 5 mL/kg),once a day.Sham operation group and model group were given 5 mL/kg normal saline intragastric administration,once a day.Each group was treated for 7 days.Neurological deficit score was evaluated by modified neurological deficit scoring standard(mNSS),cerebral hematoma volume was calculated by Tada formula,brain water content was measured by weightlessness method,blood brain barrier(BBB)permeability was measured by Evans blue(EB)method.Ultrastructural changes of BBB were observed by transmission electron microscopy(TEM).The expression of AQP4 and NKCC1 in brain tissues were detected by immunofluorescence and Western Blot.Results The neurological deficit score,cerebral hematoma volume,cerebral water content,and BBB permeability in model group were significantly higher than those in the sham operation group(P<0.01).The neurological deficit score,cerebral hematoma volume,and BBB permeability of AND50 mg/kg group and nimodipine 10 mg/kg groups were significantly lower than those of the model group(P<0.01).The neurological deficit score,cerebral hematoma volume,cerebral water content,and BBB permeability in AND50 mg/kg group were significantly lower than those in the nimodipine 10 mg/kg group(P<0.05 or P<0.01).TEM observation showed that the basal membrane of model group was uneven,enlarged,broken,tight junction structures(

关 键 词：脑出血 脑水肿 穿心莲内酯 血脑屏障 水通道蛋白4 钠-钾-氯协同转运蛋白1 

分 类 号：R74[医药卫生—神经病学与精神病学]