摘要
目的 探讨精胺后处理保护脑缺血/再灌注损伤(I/RI)小鼠模型的疗效及其分子机制。方法 60只6~8 w龄C57BL小鼠随机分为6组(每组10只):(1)对照(Control)组;(2)假手术(Sham)组;(3)模型(MCAO)组;(4)模型+精胺治疗(MCAO+Spm)组;(5)MCAO+Spm+Adv shRNA-Drp-1组;(6)MCAO+Spm+Adv shRNA-NT组。构建大脑中动脉闭塞(MCAO)小鼠模型。TTC(2,3,5-氯化三苯基四氮唑)染色脑切片测定脑梗死体积。尼氏(Nissl)染色脑组织切片测定神经元死亡情况。ELISA测定活性氧自由基(ROS)、超氧化物歧化酶(SOD)和丙二醛(MDA)水平。腺病毒转染敲低脑组织动态相关蛋白-1 (Drp-1)。Western blot测定脑组织Drp-1、B淋巴细胞瘤基因-2相关X蛋白(Bax)、B淋巴细胞瘤基因-2(Bcl-2)、PTEN诱导激酶1(PINK1)和自噬相关蛋白(Parkin-1)的表达情况。结果 与Sham组相比,MCAO组脑梗死体积所占百分比升高(P<0.05),Nissl阳性细胞所占百分比降低(P<0.05),ROS和MDA水平升高(P<0.05),SOD水平降低(P<0.05),Drp-1、Bax、PINK1和Parkin-1的表达水平升高(P <0.05),Bcl-2表达水平降低(P<0.05)。与MCAO组相比,MCAO+Spm组脑梗死体积所占百分比降低(P<0.05),Nissl阳性细胞所占百分比增多(P <0.05),ROS和MDA水平降低(P <0.05),SOD水平升高(P <0.05),Drp-1、Bcl-2、PINK1和Parkin-1的表达水平升高(P<0.05),Bax的表达水平降低(P<0.05)。与MCAO+Spm组相比,MCAO+Spm+Adv shRNA-Drp-1组Drp-1和Bax表达水平降低,Bcl-2、PINK1和Parkin-1的表达水平升高(P <0.05)。结论 Spm后处理通过Drp-1上调线粒体自噬发挥保护I/RI损伤小鼠模型的功效。
Objective To investigate the protective effect and molecular mechanism of spermine post-conditioning on cerebral ischemia/reperfusion injury(I/RI).Methods Sixty 6-8 week-old C57BL mice were randomly divided into 6 groups(n=10).①Control group;②Sham group;③MCAO group;④MCAO+Spm group;⑤MCAO+Spm+Adv shRNA-Drp-1 group;(6)MCAO+Spm+Adv shRNA-NT group.Middle cerebral artery occlusion(MCAO)mice model was constructed.Infarct volume was measured by TTC staining.Nissl staining was used to measure neuronal death.ELISA was used to determine the levels of reactive oxygen species(ROS),superoxide dismutase(SOD)and malondialdehyde(MDA)in brain tissues.Adenovirus transfection was used to knock-down dynamin-1-like protein(Drp-1)in brain tissues.Western blot was used to determine the expressions of Drp-1,BCL2 associated X(Bax),Bcl-2,PTEN induced kinase 1(PINK1)and Parkin-1 in brain tissues.Results Compared with Sham group,the percentage of cerebral infarction volume in MCAO group was increased(P<0.05),with the percentage of Nissl positive cells decreased(P<0.05).The levels of ROS and MDA were increased(P<0.05),with the level of SOD decreased(P<0.05).The expression levels of Drp-1,Bax,PINK1 and Parkin-1 were increased(P<0.05),with the expression level of Bcl-2 decreased(P<0.05).Compared with MCAO group,the percentage of cerebral infarction volume in MCAO+Spm group was decreased(P<0.05),with the percentage of Nissl positive cells increased(P<0.05).The levels of ROS and MDA were decreased(P<0.05),with the level of SOD increased(P<0.05).The expression levels of Drp-1,Bcl-2,PINK1 and Parkin-1 were increased(P<0.05),with the expression level of Bax decreased(P<0.05).Compared with MCAO+Spm group,the expression levels of Drp-1 and Bax in MCAO+Spm+Adv shRNA-Drp-1 group were decreased(P<0.05),with the expression levels of Bcl-2,PINK1 and Parkin-1 increased(P<0.05).Conclusion Spm post-condioning protected cerebral I/RI mice model by interacting with Drp-1 and up-regulating mitophagy.
作者
李偲
冷光朋
Li Si;Leng Guangpeng(Department of Rehabilitation Medicine,Liyuan Hospital Afiliated to Tongji Medical College,Huazhong University of Science and Technology,Wuhan Hubei Province 430070,China)
出处
《脑与神经疾病杂志》
CAS
2024年第6期371-376,共6页
Journal of Brain and Nervous Diseases
基金
广西壮族自治区卫生健康委员会青年基金项目(Z20200050)。
