摘要
目的研究清脑益元汤对大鼠脑缺血损伤后线粒体融合蛋白2表达的影响,探究清脑益元汤对缺血性脑损伤神经元保护作用的部分机制。方法将96只健康SD大鼠按随机数字表法分为4组:空白组(n=6)、假手术组(n=30)、模型组(n=30)、清脑益元汤组(n=30),除空白组外,假手术组、模型组及清脑益元组按缺血损伤后1 d、3 d、7 d、14 d、28 d五个取材时间点分为5个亚组。预给药7 d后,采用改良的Longa线栓法制备大鼠急性脑缺血损伤模型,造模成功后取大鼠缺血侧皮质区脑组织,采用免疫蛋白印迹(Western blot)和实时荧光定量PCR(Real-Time PCR)法检测大鼠缺血侧皮质区Mfn2蛋白及mRNA表达。结果(1)神经功能缺损评分:空白组和假手术组大鼠的神经功能评分为0分,两组比较差异无统计学意义(P>0.05);与模型组相比,清脑益元组在各个时间点的神经功能缺损症状均有改善(P<0.01);(2)Western blot法:空白组和假手术组Mfn2蛋白表达水平在同一时间点比较无明显差异(P>0.05);与假手术组同一取材时间点相比,模型组Mfn2蛋白表达水平降低(P<0.01);与模型组同一取材时间点比较,清脑益元汤组Mfn2蛋白表达水平显著上升(P<0.01);(3)Real-Time PCR法:空白组和假手术组Mfn2mRNA表达水平在同一时间点比较无明显差异(P>0.05);与假手术组同一取材时间点相比,模型组Mfn2mRNA表达水平显著降低(P<0.01);与模型组同一取材时间点相比,清脑益元汤组Mfn2mRNA表达水平升高(P<0.01)。结论清脑益元汤可通过上调脑缺血损伤后Mfn2表达,抑制线粒体损伤,减少神经细胞凋亡,进而发挥脑保护的作用。
Objective To study the effect of Qingnao Yiyuan Decoction(清脑益元汤)on the expression of mitochondrial fusion protein 2(Mfn2)after cerebral ischemic injury in rats and explore the protective mechanism of Qingnao Yiyuan Decoction on neurons after ischemic brain injury.Methods Ninety-six healthy SD rats were randomly divided into 4 groups:blank group(n=6),sham operation group(n=30),model group(n=30)and Qingnao Yiyuan Decoction group(n=30).The sham operation group,model group and Qingnao Yiyuan Decoction group were divided into 5 subgroups according to the time points of 1 d,3 d,7 d,14 d and 28 d after ischemic injury.After 7 days of preadministration,the rat model of acute cerebral ischemic injury was established by modified Longa thread embolism method.After the model was successfully established,the brain tissue of the ischemic side of the rat was taken out.The expressions of Mfn2 protein and mRNA in ischemic cortex of rats were detected by immunoblotting(Westernblot)and real-time fluorescence quantitative PCR(Real-TimePCR).Results(1)Neurological deficit score:The neurological function score of rats in the blank group and the sham operation group was 0,and there was no significant difference between the two groups(P>0.05).Compared with the model group,the symptoms of neurological deficit in Qingnao Yiyuan Decoction group were improved at all time points(P<0.01).(2)Westernblot method:There was no significant difference in the expression of Mfn2 protein between the blank group and the sham operation group at the same time point,but the expressionof Mfn2 protein in the model group was lower than that in the sham operation group at the same time point.Compared with the model group at the same time point,the expression of Mfn2 protein in Qingnao Yiyuan Decoction group was significantly higher than that in the model group(P<0.01).(3)Real-TimePCR method:There was no significant difference in the expression of Mfn2 mRNA between the blank group and the sham operation group at the same time point,but the Mfn2 mRNA ex
作者
俞睿
祝美珍
向昱臻
刘倩菁
苏萍
韦亮
孙诗杰
YU Rui;ZHU Meizhen;XIANG Yuzhen;LIU Qianjing;SU Ping;WEI Liang;SUN Shijie(Guangxi University of Traditional Chinese Medicine,Nanning 530200,Guangxi,China)
出处
《辽宁中医杂志》
CAS
2021年第1期174-178,共5页
Liaoning Journal of Traditional Chinese Medicine
基金
国家自然科学基金(81460725)
广西特色实验动物病证模型重点实验室项目(J14049)
广西一流学科建设重点项目(2018XK004)
广西中医药大学研究生科研创新重点项目课题(YCSZ2019003)。
关键词
脑缺血
清脑益元汤
线粒体
线粒体融合蛋白2
cerebral ischemia
Qingnao Yiyuan Decoction(清脑益元汤)
mitochondria
mitochondrial fusion protein 2
