机构地区:[1]山西大同大学脑科学研究所,山西大同037009 [2]山西中医药大学国家中医药管理局多发性硬化益气活血重点研究室/神经生物学研究中心,山西晋中030619 [3]山西医科大学生理学系/第一临床医学院,山西太原030001 [4]大同市第四人民医院,山西大同037008
出 处:《细胞与分子免疫学杂志》2023年第8期701-707,共7页Chinese Journal of Cellular and Molecular Immunology
基 金:山西省高等学校科技创新项目(2020L0484);山西省基础研究计划(20210302123476,20210302123478);山西省卫健委医学科技领军团队(2020TD05);基于炎性反应的重大疾病创新药物山西省重点实验室开放课题(202105D121011);山西中医药大学青年科学家培育项目(2021-PY-QN-09)。
摘 要:目的 探讨敲低Rho相关螺旋卷曲激酶2(ROCK2)基因对淀粉样前体蛋白/早老素1(APP/PS1)双转基因小鼠认知功能的影响及其机制。方法 APP/PS1双转基因小鼠随机分为AD模型组(AD组)、 ROCK2基因敲低组(shROCK2组)、 ROCK2基因敲低对照组(shNC组),同龄野生型C57BL/6小鼠作为野生型对照(WT组)。Morris水迷宫和Y迷宫实验测试小鼠认知功能,尼氏染色检测神经元形态,免疫荧光组织化学染色检测磷酸化动力蛋白相关蛋白1(p-Drp1)和线粒体融合蛋白1(Mfn1)的表达,Western blot法检测海马组织ROCK2、裂解型胱天蛋白酶3(c-caspase-3)、 B细胞淋巴瘤因子2(Bcl2)、 Bcl2相关蛋白X(BAX)、 p-Drp1、线粒体分裂蛋白1(Fis1)、视神经萎缩因子1(OPA1)、 Mfn1和Mfn2的蛋白表达。结果 与AD组小鼠相比,shROCK2组小鼠ROCK2表达明显减少;其认知功能明显改善,海马CA3和DG区神经元数量增加,尼氏体染色深;c-caspase-3和BAX表达降低,同时Bcl2表达升高;线粒体分裂相关蛋白p-Drp1和Fis1的表达减少,而线粒体融合相关蛋白OPA1、 Mfn1和Mfn2的表达增加。结论 敲低ROCK2可以显著改善APP/PS1小鼠的认知功能,并抑制神经细胞凋亡,其机制可能与促进线粒体融合并抑制其分裂有关。Objective To explore the effect of knocking down Rho-associated coiled-coil kinase(ROCK2)gene on the cognitive function of amyloid precursor protein/presenilin-1(APP/PS1)double transgenic mice and its mechanism.Methods APP/PS1 double transgenic mice were randomly divided into AD model group(AD group),ROCK2 gene knock-down group(shROCK2 group),ROCK2 gene knock-down control group(shNCgroup),and wild-type C57BL/6 mice of the same age served as the wild-type control(WT group).Morris water maze and Y maze were employed to test the cognitive function of mice.Neuron morphology was detected by Nissl staining.Immunofluorescence histochemical staining was used to detect the expression of phosphorylated dynamin-related protein 1(p-Drp1)and mitochondrial fusion 1((Mfn1).Western blot analysis was used to detect the expression ROCK2,cleaved-caspase-3(c-caspase-3),B-cell lymphoma 2(Bcl2),Bcl2-related protein X(BAX),p-Drpl,mitochondrial fission 1(Fis1),optic atrophy 1(OPA1),Mfn1 and Mfn2.Results Compared with AD group mice,the expression of ROCK2 in shROCK2 group mice was significantly reduced;the cognitive function was significantly improved with the number of neurons in the hippocampal CA3 and DG areas increasing,and nissl bodies were deeply stained;the expression of c-caspase-3 and BAX was decreased,while the expression of Bcl2 was increased;the expression of mitochondrial division related proteins p-Drpl and Fis1 were decreased,while the expression of mitochondrial fusion-related proteins OPA1,Mfn1 and Mfn2 were increased.Conclusion Knock-down of ROCK2 gene can significantly improve the cognitive function and inhibit the apoptosis of nerve cells of APP/PS1 mice.The mechanism may be related to promoting mitochondrial fusion and inhibiting its division.
关 键 词:阿尔茨海默病 淀粉样前体蛋白/早老素1(APP/PS1)小鼠 Rho相关螺旋卷曲激酶2(ROCK2) 认知障碍 凋亡 线粒体动力学
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