摘要
目的探讨miR-26抑制帕金森患者泛素-蛋白酶体系统(UPS)对含α突触核蛋白(α-Syn)降解作用。方法将120只成年雄性C57BL/6小鼠随机分为对照组与模型组,每组各60只。通过1-甲基-4-苯基-1,2,3,6-四氢吡啶(MPTP)法建立帕金森病小鼠模型,采用反转录酶-聚合酶链锁反应(RT-PCR)检测模型组小鼠中脑黑质区miR-26和α-Syn的mRNA水平,并分析二者的相关性。通过1-甲基-4-苯基-吡啶离子(MPP+)法建立帕金森病细胞模型,RT-PCR检测miR-26的表达水平,并转染miR-26模拟物或miR-26抑制物或加入MG132,RT-PCR和蛋白免疫印迹法检测α-Syn表达水平。结果RT-PCR检测结果显示,模型组小鼠中脑黑质区组织中miR-26、α-Syn水平均显著高于对照组,两组间比较,差异有统计学意义(P<0.05)。模型组小鼠脑黑质区组织中miR-26水平与α-Syn的mRNA水平呈正相关(r=0.7622,P<0.05)。蛋白免疫印迹法和RT-PCR检测结果显示,SH-SY5Y细胞在转染miR-26模拟物后,α-Syn的蛋白水平升高;转染miR-26抑制物后,α-Syn的蛋白水平降低;而转染miR-26模拟物或miR-26抑制物后,α-Syn的mRNA水平均无显著变化;加入MG132后,α-Syn蛋白水平升高,α-Syn的mRNA水平无显著变化。结论miR-26水平在帕金森病小鼠模型中上调,其不能直接调节α-Syn的表达,但能抑制UPS对α-Syn蛋白的降解。
Objective To investigate the inhibition of miR-26 on the degradation ofα-Synuclein(α-Syn)by the ubiquitin-proteasome system(UPS)in patients with Parkinson′s disease.Methods A total of 120 adult male C57BL/6 mice were randomly divided into control group and model group,with 60 mice in each group.The mouse model of Parkinson′s disease was established by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine(MPTP)method.The mRNA levels of miR-26 andα-Syn in the substantia nigra of mice in the model group were detected by reverse transcription-polymerase chain reaction(RT-PCR),and the correlation between the two was analyzed.The cell model of Parkinson′s disease was established by 1-methyl-4-phenyl-pyridine ino(MPP+)method,the expression level of miR-26 was detected by RT-PCR.The miR-26 mimics or miR-26 inhibitors were transfected or MG132 was added.The expression levels ofα-Syn were detected by RT-PCR and Western blot.Results The results of RT-PCR showed that the levels of miR-26 andα-Syn in the substantia nigra of the midbrain of mice in the model group were significantly higher than those in the control group,and the differences between the two groups were statistically significant(P<0.05).There was a positive correlation between miR-26 level andα-Syn mRNA level in the substantia nigra of mice in model group(r=0.7622,P<0.05).Western blot and RT-PCR results showed that the protein level ofα-Syn in SH-SY5Y cells was increased after transfection with miR-26 mimics.The protein level ofα-Syn reduced after transfection with miR-26 inhibitor.However,there was no significant change in the mRNA level ofα-Syn after transfection with miR-26 mimics or miR-26 inhibitors.After adding MG132,theα-Syn protein level increased,but theα-Syn mRNA level had no significant change.Conclusion The level of miR-26 is up-regulated in the Parkinson′s disease mouse model,which could not directly regulate the expression ofα-Syn,but could inhibit the degradation ofα-Syn by UPS.
作者
穆军山
崔晓萍
周贺
王魁花
叶建新
MU Jun-shan;CUI Xiao-ping;ZHOU He;WANG Kui-hua;YE Jian-xin(Department of Neurology,The 900th Hospital of the Joint Logistics Support Force of the Chinese People′s Liberation Army,Fuzhou 350025,China)
出处
《创伤与急危重病医学》
2021年第2期124-127,共4页
Trauma and Critical Care Medicine
基金
2017年福建省自然科学基金科技项目计划(2017J01319)。
作者简介
第一作者:穆军山(1965-),男,山东莱州人,主任医师,硕士;通信作者:叶建新,E-mail:13959199189@139.com。
