小胶质细胞中小泛素样修饰蛋白特异性蛋白酶3参与小鼠光化学栓塞法缺血性脑卒中早期的疾病进展

杜娟娟 叶蓁 周涛 任艳华 廖敏

解剖学报 ›› 2021, Vol. 52 ›› Issue (4) : 548-553.

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解剖学报 ›› 2021, Vol. 52 ›› Issue (4) : 548-553. DOI: 10.16098/j.issn.0529-1356.2021.04.008
神经生物学

小胶质细胞中小泛素样修饰蛋白特异性蛋白酶3参与小鼠光化学栓塞法缺血性脑卒中早期的疾病进展

  • 杜娟娟 叶蓁 周涛 任艳华 廖敏*
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Small ubiquitin-like modifier proteins specific protease 3 in microglia participating in the early progression of photothrombosic ischemic stroke mice model

  • DU Juan-juan  YE Zhen  ZHOU Tao  REN Yan-hua  LIAO Min*
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摘要

目的  探讨小泛素样修饰蛋白特异性蛋白酶3(SENP3)在光化学栓塞法缺血性脑卒中小鼠小胶质细胞中的表达变化,以及与光化学法栓塞缺血性脑卒中疾病进展的关系。  方法  实验小鼠分对照组、缺血性脑卒中1 d组和缺血性脑卒中7 d组(每组3只),应用Western blotting检测各组纹状体诱导型一氧化氮合酶(iNOS)、精氨酸酶1(ARG-1)和SENP3的表达和c-Jun氨基末端激酶(JNK)磷酸化水平;应用免疫荧光双标法检测小鼠纹状体小胶质细胞中iNOS和ARG-1的表达。   结果  与对照组相比,缺血性脑卒中1 d组SENP3的表达与JNK的磷酸化水平均显著上升,同时M1型小胶质细胞的标志物iNOS的表达水平显著上升;缺血性脑卒中7 d组M2型小胶质细胞的标记物ARG-1的表达显著增高。纹状体小胶质细胞特异性抗体1(Iba1)与iNOS、Iba1与ARG-1的免疫荧光双标结果与免疫印迹结果一致。   结论  光化学栓塞法缺血性脑卒中早期,小胶质细胞SENP3表达增加,进而影响JNK磷酸化的水平和小胶质细胞的极化,促进大脑炎症反应,参与缺血性脑卒中的疾病进展。

Abstract

Objective  To investigate the expression of small ubiquitin-like modifier proteins specific protease 3 (SENP3) in microglia of mice with ischemic stroke and its relationship with the progression of ischemic stroke.    Methods  The experimental animals were divided into control group, ischemic stroke day 1 group and ischemic stroke day 7 group(3 mice per group). The expression of inducible nitric oxide synthase (iNOS), argniase-1 (ARG-1), SENP3 and c-Jun N-terminal kinase (JNK) phosphorylation levels in the striatum were detected by immunoblotting. The expression of iNOS and ARG-1 in mouse striatum microglia was detected by immunofluorescence double labeling.    Results  Compared with the control group, the expression of SENP3 and the phosphorylation level of JNK in the ischemic stroke group increased significantly, and the expression of the marker iNOS of M1 type microglia increased significantly. The expression of the marker ARG-1 of M2 type microglia increased significantly in the day 7 group of ischemic stroke. The immunofluorescence doublelabeled result  of striatum ionized calcium binding adapter molecule 1 (Iba1) and iNOS, Iba1 and ARG-1 were consistent with the result  of immunoblotting.    Conclusion  In the early stage of ischemic stroke, the expression of SENP3 in microglia increases, which promote the cerebral inflammatory response by affecting the level of JNK phosphorylation and the polarization of microglia, and participate in the progression of ischemic stroke.

关键词

光化学栓塞法缺血性脑卒中 / 小胶质细胞 / 小泛素样修饰蛋白特异性蛋白酶3 / 免疫印迹法 / 小鼠

Key words

Photothrombosic ischemic stroke / Microglia / Small ubiquitin-like modifier proteins specific protease 3 / Western blotting / Mouse

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杜娟娟 叶蓁 周涛 任艳华 廖敏. 小胶质细胞中小泛素样修饰蛋白特异性蛋白酶3参与小鼠光化学栓塞法缺血性脑卒中早期的疾病进展[J]. 解剖学报. 2021, 52(4): 548-553 https://doi.org/10.16098/j.issn.0529-1356.2021.04.008
DU Juan-juan YE Zhen ZHOU Tao REN Yan-hua LIAO Min. Small ubiquitin-like modifier proteins specific protease 3 in microglia participating in the early progression of photothrombosic ischemic stroke mice model[J]. Acta Anatomica Sinica. 2021, 52(4): 548-553 https://doi.org/10.16098/j.issn.0529-1356.2021.04.008
中图分类号: R329.2    

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基金

温州市基础性科研项目;人类肠道共生细菌在LPS诱导的帕金森模型小鼠中的作用及机制研究

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