丹参酮ⅡA对缺氧缺血性脑病新生大鼠海马组织中miR-132表达的调节作用及其机制  

Effects of tanshinone ⅡA on miR-132 expression in hippocampus tissue of neonatal rats with hypoxic-ischemic encephalopathy and its mechanism

作　　者：张洋[1] 陈美月 崔莹[1] 刘娜[1] ZHANG Yang;CHEN Meiyue;CUI Ying;LIU Na(Department of Pediatrics,Second Department of First Hospital,Jilin University,Changchun 130031,China)

机构地区：[1]吉林大学第一医院二部儿科,吉林长春130031

出　　处：《吉林大学学报（医学版）》2022年第4期905-914,共10页Journal of Jilin University：Medicine Edition

基　　金：吉林省科技厅卫生科研人才计划项目(2019SCZ045)。

摘　　要：目的:探讨丹参酮ⅡA(TanⅡA)对新生大鼠缺氧缺血性脑病的干预作用,并阐明其作用机制。方法:40只SD大鼠随机分为假手术组、模型组、TanⅡA组和miR-132抑制剂(miR-132 antagomir)组,每组10只。大鼠双重结扎颈总动脉并放入缺氧舱建立缺血缺氧性脑病模型。TanⅡA组大鼠造模后腹腔注射TanⅡA注射液(24 mg·kg^(-1)),双侧脑室注射2μg miR-132阴性对照质粒(NC);假手术组和模型组大鼠腹腔注射等量生理盐水;miR-132 antagomir组大鼠造模后腹腔注射TanⅡA注射液(24 mg·kg^(-1)),双侧脑室注射2μg miR-132 antagomir;各组大鼠连续注射7 d。实时荧光定量PCR(RT-qPCR)法和荧光原位杂交检测各组大鼠海马组织中miR-132表达水平和表达强度,采用改良神经功能缺失评分评价各组大鼠神经功能,HE染色观察各组大鼠海马组织病理形态表现,TUNEL法检测各组大鼠海马组织中神经元细胞凋亡率,酶联免疫吸附测定(ELISA)法检测各组大鼠海马组织中单胺类神经递质水平,高尔基染色检测各组大鼠海马组织中树突棘密度,Western blotting法检测各组大鼠海马组织中突触后致密物95(PSD-95)、生长相关蛋白43(GAP-43)、微管相关蛋白2(MAP-2)和脑源性神经营养因子(BDNF)蛋白表达水平。结果:与假手术组比较,模型组大鼠海马组织中miR-132表达水平和表达强度明显降低(P<0.05);与模型组比较,TanⅡA组大鼠海马组织中miR-132表达水平和表达强度明显升高(P<0.05);与TanⅡA组比较,miR-132 antagomir组大鼠海马组织中miR-132表达水平和表达强度明显降低(P<0.05)。与假手术组比较,模型组大鼠神经功能缺失评分明显升高(P<0.05);与模型组比较,TanⅡA组大鼠神经功能缺失评分明显降低(P<0.05);与TanⅡA组比较,miR-132 antagomir组大鼠神经功能缺失评分明显升高(P<0.05)。HE染色,假手术组大鼠海马组织无损伤;与假手术组比较,模型组大鼠海马组织损伤严重;与模型组比较,Objective:To investigate the intervention effect of tanshinone ⅡA(TanⅡA)in the neonatal rats with hypoxic-ischemic encephalopathy,and to clarify its mechanism.Methods:Forty SD rats were randomly divided into sham operation group,model group,TanⅡA group and miR-132 antagomir group,with 10 rats in each group.The rat models of hypoxic-ischemic encephalopathy were established by double ligation of the common carotid artery and being put into anoxic chamber.The rats in TanⅡA group were intrapeitoneally injected with TanⅡA injection(24 mg·kg^(-1)),and 2μg miR-132 negative control(NC)was injected into the bilateral ventricles.The rats in sham operation group and model group were intraperitoneally injected with the same amount of normal saline.After modeling,the rats in miR-132 antagomir group were intraperitoneally injected with tanshinoneⅡA injection(24 mg·kg^(-1)),and 2μg miR-132 antagomir was injected into the bilateral ventricles;the tars in various groups were injected for 7 d.The expression levels and fluorescence intensities of miR-132 in hippocampus tissue of the rats in various groups were detected by real-time fluorescence quantitative PCR(RT-qPCR)and fluorescence in situ hybridization.Modified neurological deficit score was used to evaluate the neural function of the rats in various groups.HE staining was used to observe the pathomorphology of hippocampus tissue of the rats in various groups.TUNEL assay was used to detect the apoptotic rates of neurons in hippocampus tissue of the rats in various groups.The levels of monoamine neurotransmitters in hippocampus tissue of the rats in various groups were detected by enzyme-linked immunosorbent assay(ELISA).The densities of dendritic spines in hippocampus tissues of the rats in various groups were detected by Golgi staining.The protein expression levels of postsynaptic density-95(PSD-95),growth associated protein-43(GAP-43),microtubule-associated protein-2(MAP)-2 and brain-derived neurotrophic factor(BDNF)in hippocampus tissue of the rats in variou

关 键 词：丹参酮ⅡA磺酸钠 缺氧缺血性脑病 微小RNA-132 树突棘密度 突触前蛋白 

分 类 号：R-332[医药卫生] R748