基于微小RNA-27a-3p/磷酸二酯酶3A探讨益气活血方对缺血性脑梗死小鼠微血管保护的分子机制  

Investigation on molecular mechanism of Yiqi Huoxue Recipe on microvascular protection in mice model with ischemic cerebral infarction based on micro RNA-27a-3p/phosphodiesterase 3A

作　　者：欧阳方 张小红[2] Ouyang Fang;Zhang Xiaohong(Wuhan First Hospital(Wuhan Hospital of Traditional Chinese and Western Medicine),Wuhan 430022,Hubei,China;the Sixth Hospital of Wuhan,Wuhan 430022.Hubei,China)

机构地区：[1]武汉市第一医院(武汉市中西医结合医院),湖北武汉430022 [2]武汉市第六医院,湖北武汉430022

出　　处：《中国中西医结合急救杂志》2020年第6期744-748,共5页Chinese Journal of Integrated Traditional and Western Medicine in Intensive and Critical Care

基　　金：湖北省武汉市医药卫生科研项目(WX16E07)。

摘　　要：目的观察益气活血方对缺血性脑梗死模型小鼠脑微血管的保护作用,并探讨其分子机制.方法选择50只成年雄性SPF C57小鼠,按随机数字表法分为假手术组、永久性大脑中动脉闭塞(pMCAO)模型组和益气活血方高、中、低剂量组,每组10只.采用改良的线栓法复制pMCAO卒中模型;假手术组不进行栓线操作.术后假手术组、pMCAO模型组均给予相同体积的生理盐水;益气活血方(组成:黄芪40 g、人参15 g、丹参30 g、益母草20 g、茯苓25 g、葶苈子10 g,由河北中医学院中药制剂中心制备成颗粒剂)高、中、低剂量组分别按240、120、60 mg/kg剂量灌胃.观察各组小鼠脑组织微血管形态、脑梗死体积的变化、血管内皮生长因子(VEGF)、血管生成素-1(Ang-1)、内皮素-1(ET-1)含量以及微小RNA(miRNA)-27a-3p的mRNA与磷酸二酯酶3A(PDE3A)的mRNA和蛋白表达水平.结果pMCAO模型组脑梗死体积较假手术组明显增大〔(60.25±12.05)%比(2.02±0.26)%,P<0.01〕;各给药组脑梗死体积均较pMCAO模型组明显减小,且以益气活血方高剂量组的减小程度较低剂量和中剂量组更显著〔(38.98±10.26)%比(50.26±11.44)%、(45.77±8.95)%〕.光镜下可见:假手术组脑血管排列规则,管腔血液充盈,血管周围红润;pMCAO模型组梗死区血管腔内红细胞呈线性排列,管周间隙加大并有严重水肿,半暗区血管周围出现炎性细胞浸润;益气活血方各剂量组脑微血管受损程度均较pMCAO组明显减轻.pMCAO模型组VEGF、Ang-1、ET-1含量及PDE3A的mRNA和蛋白表达水平均较假手术组明显升高〔VEGF(ng/L):6.44±1.26比1.44±0.59,Ang-1(ng/L):9.68±2.33比2.33±1.02,ET-1(ng/L):50.55±18.66比20.44±9.56,PDE3A mRNA(2^(-ΔΔCt)):6.99±0.86比1.00±0.00,PDE3A蛋白(PDE3A/GAPDH):2.84±1.49比1.00±0.00,均P<0.01〕,miR-27a-3p mRNA表达较假手术组明显降低(2^(-ΔΔCt):0.36±0.06比1.00±0.00);益气活血方高、中、低剂量组VEGF、Ang-1、ET-1含量及PDE3A的mRNA和蛋白�Objective To observe the protective effect of Yiqi Huoxue Recipe(YHR)on cerebral miero-vessels of ischemic cerebral infarction mice model and explore its molecular mechanism.Methods According to random number table method,the 50 adult male SPF C57 mice were divided into sham operation group,permanent middle cerebral artery ocelusion(pMCAO)model group and high,medium and low dose YHR groups,with 10 mice in each group.The modified thread bolt method was used to replicate the pMCAO stroke model.In the sham operated group,similar operation without suture management was performed.After operation,the sham operated group and pMCAO model group were given the same volume of normal saline;in high,medium,low dose Y HR groups,YHR(ingredients:Radix Astragali 40 g,Panax Salviae Miltiorrhizae 30 g,Herba Leonuri 20 g.Poria 25 g.Semen ginseng 15 g,Radix Lepidii 10 g.were mixed and prepared as granules by Traditional Chinese Medieine Preparation Center of Hebei University of Traditional Chinese Medicine)high-,mid-and low-dose groups were given with the doses of 240,120 and 60 mg/kg by gavage administration.The morphological changes of cerebral micro-vessels,volume of cerebral infarction,contents of vascular endothelial growth factor(V ECF),angiopoietin-1(Ang-1)and endothelin-1(ET-1),levels of mRNA and protein expressions of miRNA-27a-3p and phosphodiesterase 3A(PDE3A)were observed.Results Compared with the sham operated group,the cerebral infarction volume in pMCAO model group was signifieantly increased[(60.25±12.05)%vs.(2.02±0.26)%,P<0.01];and the cerebral infarction volumes in each drug administration group was significantly lower than that in pMCAO model group,and the degree of reduction in high dose YHR group was more significant than those in low and medium dose YHR groups[(38.98±10.26)%vs.(50.26±11.44)%.(45.77±8.95)%].Under the light microscope,the cerebral vessels in sham operated group were arranged regularly,the lumen of the vessels was flled with blood and vascular surroundings were ruddy;it could be seen that the

关 键 词：缺血性脑梗死 微血管 益气活血方 脑梗死 

分 类 号：R28[医药卫生—中药学]