基质金属蛋白酶组织抑制因子1通过丝裂原活化蛋白激酶途径促进小胶质细胞M2极化，以改善缺血后的早期脑损伤。

TIMP1 promotes microglia M2 polarization through MAPK pathway to ameliorate early brain injury after ischemia.

作者信息

Zhao Kangkang, Huang Zizhao

机构信息

Department of Neurosurgery, Yulin Hospital, The First Hospital Affiliated to Xi'an Jiaotong University, Yulin, 719000, Shaanxi, China.

出版信息

Hereditas. 2025 Jul 2;162(1):119. doi: 10.1186/s41065-025-00491-8.

DOI:10.1186/s41065-025-00491-8

PMID:40604895

Abstract

OBJECTIVE

To explore the molecular regulatory mechanisms and biomarkers in regulating early brain injury (EBI) and inflammatory response after ischemic stroke (IS).

METHODS

Gene expression profiles of GSE148350, GSE35338, and GSE58294 were analyzed to screen the core genes involved in EBI after IS. Middle cerebral artery occlusion and reperfusion (MCAO/R) model and oxygen and glucose deprivation and reoxygenation (OGD/R) model were used to construct in vivo and in vitro IS models. In MCAO/R model, the effects of tissue inhibitor of metalloproteinase-1 (TIMP1) were investigated by Zea longa score, brain water content assessment and histological analysis. In OGD/R model, after TIMP1 was overexpressed in BV2 cells, M1 and M2 polarization markers (iNOS and Arg-1) in BV2 cells were detected by Western blot, and the effects of BV2 on the viability and apoptosis of HT22 cells were evaluated by cell counting kit-8 and flow cytometry, respectively. Additionally, the effects of TIMP1 overexpression on MAPK pathway in BV2 cells were also detected by Western blot.

RESULTS

Two core genes, TIMP1 and vimentin (VIM) were screened from 254 differentially expressed genes in IS. TIMP1 was closely associated with the dysregulation of immune cell infiltration. TIMP1 overexpression significantly mitigated MCAO/R-induced neurological dysfunction, brain edema, neuronal apoptosis and inflammatory response in rats. In vitro, it was revealed that TIMP1 overexpression in BV2 cells increased viability and inhibited apoptosis of HT22 cells. In BV2 cells, TIMP1 overexpression promoted the expression of Agr-1 and inhibited the expression of iNOS. In addition, overexpression of TIMP1 inhibited OGD/R-induced increases in the phosphorylation levels of p38, JNK and ERK proteins in BV2 cells.

CONCLUSION

This study identified a post-IS EBI regulator, TIMP1. TIMP1 promotes M2 polarization of microglia and ameliorate neurological injury after IS by inactivating MAPK signaling pathway.

摘要

目的

探讨调节缺血性脑卒中（IS）后早期脑损伤（EBI）及炎症反应的分子调控机制和生物标志物。

方法

分析GSE148350、GSE35338和GSE58294的基因表达谱，以筛选IS后EBI相关的核心基因。采用大脑中动脉闭塞再灌注（MCAO/R）模型和氧糖剥夺再复氧（OGD/R）模型构建体内和体外IS模型。在MCAO/R模型中，通过Zea longa评分、脑含水量评估和组织学分析研究金属蛋白酶组织抑制剂-1（TIMP1）的作用。在OGD/R模型中，在BV2细胞中过表达TIMP1后，通过蛋白质免疫印迹法检测BV2细胞中M1和M2极化标志物（iNOS和Arg-1），并分别通过细胞计数试剂盒-8和流式细胞术评估BV2对HT22细胞活力和凋亡的影响。此外还通过蛋白质免疫印迹法检测TIMP1过表达对BV2细胞中MAPK通路的影响。

结果

从IS的254个差异表达基因中筛选出两个核心基因，即TIMP1和波形蛋白（VIM）。TIMP1与免疫细胞浸润失调密切相关。TIMP1过表达显著减轻MCAO/R诱导的大鼠神经功能障碍、脑水肿、神经元凋亡和炎症反应。在体外，研究发现BV2细胞中TIMP1过表达可提高HT22细胞的活力并抑制其凋亡。在BV2细胞中，TIMP1过表达促进Agr-1的表达并抑制iNOS的表达。此外，TIMP1过表达抑制OGD/R诱导的BV2细胞中p38、JNK和ERK蛋白磷酸化水平升高。