神经生长因子联合BMSCs-源外泌体通过Keap1/NQO1/Nrf2信号通路缓解脑出血大鼠神经损伤  

Nerve Growth Factor Combined with BMSCs Exosomes Alleviates Neural Injury in Rats with Intracerebral Haemorrhage through Keap1/NQO1/Nrf2 Signaling Pathway

作　　者：李方[1] 唐仕军 麦麦提依明·托合提 LI Fang;TANG Shijun;Maimaitiyiming Tuoheti(Department of Neurosurgery,Xinjiang Uygur Autonomous Region People’s Hospital,Urumqi,830001,China)

机构地区：[1]新疆维吾尔自治区人民医院神经外科,乌鲁木齐市830001

出　　处：《医学分子生物学杂志》2024年第4期341-346,共6页Journal of Medical Molecular Biology

基　　金：新疆维吾尔自治区人民医院院内项目(No.20210108)。

摘　　要：目的探讨大鼠神经生长因子(rat nerve growth factor,RtNGF)联合骨髓间充质干细胞-源外泌体(BMSCs exosomes)缓解脑出血(intracerebral haemorrhage,ICH)大鼠神经损伤的疗效及其作用机制。方法制备BMSCs exosomes。60只大鼠随机分为假手术(Sham)组,ICH组,ICH+RtNGF组,ICH+BMSCs exosomes组,ICH+RtNGF+BMSCs exosomes组,每组12只。建立ICH大鼠模型,并给予RtNGF或BMSCs exosomes单独治疗或联合治疗。制备各分组大鼠脑组织石蜡组织切片,并对其进行HE染色。qPCR法测定各分组大鼠脑组织中Keap1/NQO1/Nrf2信号通路蛋白质因子mRNA的相对表达水平。免疫组化(IHC)法测定各分组大鼠脑组织中Keap1/NQO1/Nrf2信号通路蛋白质因子的表达水平。结果与Sham组相比,ICH组大鼠脑组织中存在多处明显的组织腔隙和血凝块。ICH+RtNGF组、ICH+BMSCs exosomes组和ICH+RtNGF+BMSCs exosomes组均有所改善,且ICH+RtNGF+BMSCs exosomes组缓解神经损伤效果最好。与Sham组相比,ICH组大鼠脑组织中Keap1,NQO1和Nrf2 mRNA表达水平升高(P<0.05);上述3种蛋白质IHC相对染色评分升高(P<0.05)。与ICH组相比,ICH+RtNGF组、ICH+BMSCs exosomes组和ICH+RtNGF+BMSCs exosomes组上述检测指标水平降低(P<0.05),且与ICH+RtNGF组或ICH+BMSCs exosomes组相比,ICH+RtNGF+BMSCs exosomes组上述检测指标水平更低(P<0.05)。结论RtNGF联合BMSCs exosomes治疗ICH大鼠,可通过Keap1/NQO1/Nrf2信号通路降低氧化应激缓解神经损伤。Objective To explore the therapeutic effect and mechanisms of rat nerve growth factor(RtNGF)combined with bone marrow mesenchymal stem cell-derived exosomes(BMSCs exosomes)on neural injury of intracerebral haemorrhage(ICH)rats.Methods The BMSCs exosomes were prepared for the following experiments.Sixty rats were randomly divided into 5 groups:Sham group,ICH group,ICH+RtNGF group,ICH+BMSCs exosomes group,and ICH+RtNGF+BMSCs exosomes group,with 12 rats in each group.A ICH rat model was established and RtNGF or BMSCs exosomes were treated alone or in combination.Paraffin embedded tissue sections of rat brain tissues in each group were prepared and HE staining was performed.qPCR and Immunohistochemistry were used to determine the mRNA and protein expression levels of genes in the Keap1/NQO1/Nrf2 signaling pathway.Results Significant existence of tissue lacunes and blood clots were observed in the ICH group rather than in the Sham group.RtNGF or/and BMSCs treatments improved that situation,and the combination of RtNGF and BMSCs exosomes treatment had the best effect on alleviating neural injury.The mRNA expression levels of Keap1,NQO1 and Nrf2 in the brain tissues,and the IHC relative staining scores of the three proteins in the ICH group were increased when compared with those in the Sham group(allP<0.05).The levels of the above indicators in the ICH+RtNGF group,ICH+BMSCs exosomes group,and ICH+RtNGF+BMSCs exosomes group were decreased when compared with those in the ICH group(P<0.05),with the values the lowest in the ICH+RtNGF+BMSCs exosomes group(P<0.05).Conclusion RtNGF combined with BMSCs exosomes has significant effect on reducing oxidative stress and alleviating neural injury through Keap1/NQO1/Nrf2 signaling pathway in ICH rats.

关 键 词：脑出血 大鼠神经生长因子 BMSCs-源外泌体 Keap1/NQO1/Nrf2信号通路 神经损伤 

分 类 号：R743.34[医药卫生—神经病学与精神病学]