摘要
目的:探究阿魏酸钠(SF)通过miR-216b-3p/Nrf2通路对缺氧缺血性脑病(HIE)胚胎大鼠大脑的干预作用以及减轻氧化应激损伤的作用机制。方法:将成年雌性SD大鼠和雄鼠,按照3∶1比例合笼获得怀孕的雌鼠。然后将孕鼠分为假手术组(sham)、缺氧缺血性脑病模型组(HIE)、SF低剂量组(HIE+SF-L)和SF高剂量组(HIE+SF-H)。通过无创止血钳夹闭子宫两侧动脉和卵巢血管法制备胚胎大鼠HIE模型。SF治疗组腹膜腔注射SF。采用HE染色观察胚胎大鼠大脑皮质的病理变化;免疫荧光观察核因子红细胞系2相关因子2(Nrf2)、过氧化还原酶1(PRDX1)的表达情况;Western Blot检测胚胎大鼠脑组织中Nrf2和PRDX1蛋白的表达;real time RT-PCR检测miR-216b-3p的表达以及Nrf2和PRDX1的mRNA表达;WST-8法和TBA法检测脑组织中的超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量。结果:与sham组对比,HIE组胚胎大鼠的大脑损伤加重,病理改变明显;HIE组中Nrf2蛋白表达和mRNA水平降低(P<0.05),PRDX1的蛋白水平和mRNA水平显著上调(P<0.05),miR-216b-3p表达水平显著升高(P<0.05);并且Nrf2的平均荧光强度显著降低(P<0.05),PRDX1的平均荧光强度显著增加(P<0.05);SOD活性显著下调,MDA含量显著增加(P<0.05)。与HIE组对比,各剂量SF组的大脑皮质病理结构明显改善,脑损伤减轻;Nrf2和PRDX1的蛋白表达、mRNA水平以及平均荧光强度均显著上升(P<0.05);miR-216b-3p表达水平均显著下调,以SF高剂量组下调最为显著(P<0.05);并且SOD活性显著上调,MDA含量显著下调(P<0.05)。结论:SF通过miR-216b-3p/Nrf2信号通路在HIE的发生发展过程中发挥重要作用,并减轻胚胎大鼠受到的氧化应激损伤。
Objective:To Investigate the interventional effects of sodium ferulate(SF)on the brain of hypoxic-ischemic encephalopathy(HIE)embryonic rats via miR-216b-3p/Nrf2 pathway and the mechanism of action of SF in reducing oxidative stress damage.Method:Adult female SD rats and male rats were caged together in a 3∶1 ratio to obtain Pregnant female rats.The pregnant rats were then divided into sham group(sham),model group(HIE),SF low-dose group(HIE+SF-L)and an SF high-dose group(HIE+SF-H).The HIE model was prepared by non-invasive hemostatic forceps clamping the arteries on both sides of the uterus and the ovarian vessels.SF was injected intraperitoneally into the SF treatment group.HE staining was used to observe the pathological changes in the cerebral cortex of embryonic rats;immunofluorescence was used to observe the expression of nuclear factor-erythroid 2-related factor 2(Nrf2),and peroxiredoxin 1(PRDX1);Western Blot detection of Nrf2,and PRDX1 protein expression in embryonic rat brain tissue;real time RT-PCR to detect the expression of miR-216b-3p and the mRNA expression of Nrf2 and PRDX1;determination of superoxide dismutase(SOD)activity and malondialdehyde(MDA)content in brain tissue by WST-8 and TBA methods.Results:Compared with the Sham group,the embryonic rats in the HIE group showed increased brain damage and significant pathological changes;Nrf2 was significantly down-regulated in protein level and mRNA level in the HIE group(P<0.05),PRDX1 was significantly up-regulated in protein level and mRNA level(P<0.05),and miR-216b-3p expression levels were significantly increased(P<0.05);and the average fluorescence intensity of Nrf2 was significantly decreased(P<0.05)and the mean fluorescence intensity of PRDX1 was significantly increased(P<0.05);SOD activity was significantly down-regulated and MDA content was significantly increased(P<0.05).Compared with the HIE group,the cortical pathological structure was significantly improved and brain damage was reduced in the SF group at all doses;and protein expression,mRNA levels and mean fluorescence intensity of Nrf2 and PRDX1 were significantly increased(P<0.05);miR-216b-3p expression levels were significantly down-regulated,with the most significant down-regulation in the SF high-dose group(P<0.05);And SOD activity was significantly up-regulation and MDA content was significantly down-regulation(P<0.05).Conclusion:SF plays an important role in the development of HIE through miR-216b-3p/Nrf2 signaling pathway and reduces oxidative stress damage in embryonic rats.
作者
黄一伟
郜萌
牟宸希
徐伟靖
孙逊
陈天浩
徐辉
HUANG Yiwei;GAO Meng;MU Chenxi;XU Weijing;SUN Xu;CHEN Tianhao;XU Hui(Jiamusi University,School of Basic Medicine,Jiamusi 154007,China;Jiamusi University,Key Laboratory of Microecology-Immune Regulation Network and Related Diseases,Jiamusi 154007,China;Jiamusi University,College of Public Health,Jiamusi 154007,China)
出处
《神经解剖学杂志》
CAS
CSCD
2023年第5期529-536,共8页
Chinese Journal of Neuroanatomy
基金
黑龙江省教育厅基本科研业务费基础研究项目(2019-KYYWF-1337)。
关键词
阿魏酸钠
缺氧缺血性脑病
氧化应激
核转录因子红系2相关因子2
过氧化物还原蛋白1
大鼠
sodium ferulate(SF)
hypoxic-ischemic encephalopathy(HIE)
oxidative stress
nuclear factor-erythroid 2-related factor 2(Nrf2)
peroxiredoxin 1(PRDX1)
rat
