摘要
目的探讨血管钠肽(VNP)对小鼠脑出血的神经功能和脑水肿的影响及其潜在的作用机制。方法90只8周龄雄性C57BL/6小鼠随机分为5组:假手术组(sham组)、脑出血组(ICH组)、脑出血+血管钠肽10μg/kg处理组(ICH+10μg/kg VNP)、脑出血+血管钠肽25μg/kg处理组(ICH+25μg/kg VNP)及脑出血+血管钠肽50μg/kg处理组(ICH+50μg/kg VNP)。采用纹状体注射胶原酶Ⅶ建立脑出血小鼠模型,VNP处理组分别在造模开始前1 h及造模结束后1 h,将VNP溶液腹腔注射入小鼠体内。脑出血1,3,5,7 d应用神经功能损伤评分观察小鼠脑出血的神经功能;采用干湿重法检测脑水肿程度,用以评估VNP保护作用的最佳效应。后续实验分为三组:sham组、ICH组和ICH+25μg/kg VNP(最佳效应)组,分别通过DHE染色法观察超氧化物阴离子情况以及化学荧光检测ROS含量;应用检测试剂盒分别检测MDA含量和SOD活性评估氧化应激;利用Western blot检测内质网应激相关蛋白GRP78和CHOP的水平。结果与ICH组相比,脑出血1,3,5,7 d,10μg/kg VNP处理组的神经功能缺损评分及脑水肿程度均未明显改变(P>0.05);脑出血1 d或3 d时,25μg/kg和50μg/kg VNP处理组的神经功能缺损评分均降低(P<0.05);脑出血1 d时,25μg/kg和50μg/kg VNP处理组脑水肿程度均降低(P<0.05);脑出血3 d时,25μg/kg VNP处理组脑水肿程度显著降低(P<0.01)。因此,后续实验采用25μg/kg VNP浓度,观察脑出血3 d的指标。与ICH组相比,脑出血3 d时25μg/kg VNP组脑组织超氧化物阴离子表达受抑制,ROS及MDA含量降低(P<0.01),SOD活力提高(P<0.05),内质网应激相关蛋白GRP78和CHOP的表达下调(P<0.01)。结论VNP对小鼠脑出血损伤具有重要的神经保护作用,其潜在的机制是通过抑制氧化应激和内质网应激的过度激活,进而改善小鼠神经功能和水肿程度。
Objective To investigate the effect of vasonatrin peptide(VNP)on neurological function and cerebral edema of cerebral hemorrhage in mice and its potential mechanism.Methods Ninety eight-week-old male C57BL/6 mice were randomly divided into five groups:sham operation group(sham group),cerebral hemorrhage group(ICH group),cerebral hemorrhage+vasonatrin peptide 10μg/kg treatment group(ICH+10μg/kg VNP),cerebral hemorrhage+25μg/kg VNP treatment group(ICH+25μg/kg VNP)and cerebral hemorrhage+50μg/kg VNP treatment group(ICH+50μg/kg VNP).The mice were injected with collagenaseⅦinto the striatum to establish a mouse cerebral hemorrhage model.The mice in ICH+VNP group were injected intraperitoneally with VNP solution into the mice at 1 h before and 1 h after modeling.The neurological function and the degree of brain edema were measured at day 1,3,5,and 7 after cerebral hemorrhage by dry and wet weight method to screen the best protective effect of VNP.In sham group,ICH group and ICH+25μg/kg VNP group,DHE staining was used to observe the superoxide anion,chemical fluorescence was used to detect ROS content,the detection kit was used to detect MDA content and SOD activity,and Western blot was used to detect the levels of endoplasmic reticulum stress-related proteins GRP78 and CHOP.Results Compared with ICH group,the neurological deficit score and the degree of cerebral edema showed no significant change in 10μg/kg VNP group(P>0.05).Compared with ICH group,the neurological deficit scores were reduced at day 1 and day 3 after cerebral hemorrhage in 25μg/kg VNP group and 50μg/kg VNP group(P<0.05).Compared with ICH group,the degree of cerebral edema was reduced at day 1 after cerebral hemorrhage in 25μg/kg VNP group and 50μg/kg VNP group(P<0.05).Compared with ICH group,the degree of cerebral edema was significantly reduced at day 3 after cerebral hemorrhage in 25μg/kg VNP group(P<0.01).Therefore,25μg/kg VNP was used in the following experiment to observe the indexes at day 3 after cerebral hemorrhage.Compared with
作者
常盼
朱萧玲
于军
朱肖星
王建榜
王西辉
李静
CHANG Pan;ZHU Xiaoling;YU Jun;ZHU Xiaoxing;WANG Jianbang;WANG Xihui;LI Jing(Central Laboratory,Second Affiliated Hospital,Xi’an Medical College,Xi’an 710038,China;Department of Anesthesiology,Xijing Hospital,Air Force Military Medical University)
出处
《山西医科大学学报》
CAS
2020年第12期1360-1365,共6页
Journal of Shanxi Medical University
基金
国家自然科学基金资助项目(81670365,81870172)
陕西省科学技术厅重点研发计划-一般项目(社会发展领域)(2018SF129)
陕西省教育厅自然专项科研计划项目(20JK0900)
西安市科技局医学研究项目(20YXYJ003(3))
西安医学院第二附属医院院级课题(19KY0113)。
关键词
血管钠肽
脑出血
小鼠
神经功能
氧化应激
内质网应激
vasonatrin peptide
cerebral hemorrhage
mice
neurological function
oxidative stress
endoplasmic reticulum stress
