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The destructive role of soluble Robo4 secreted by the M1-polarized-microglia during cerebral ischemia-reperfusion in blood-brain barrier integrity

HUANG Jin-Long¹, LI Chen¹, YANG Liang-Liang¹, GAO Yang¹, ZHAO Pu-Yuan¹, YANG Zhi-Gang^1,2,*

¹Zhongshan Hospital, Fudan University, Shanghai 200032, China；²National Clinical Research Center for Interventional Medicine, Shanghai 200032, China

Abstract

This project was aimed to investigate the role and the underlying mechanism of microglia polarization on blood-brain barrier (BBB) during cerebral ischemia-reperfusion. After construction of the mouse model of cerebral ischemia-reperfusion, upregulated IL-6 and TNF-α in peripheral blood and increased IL-6 and iNOS in ischemia tissues were confirmed. The supernatant expression of TNF-α and IL-6, as well as IL-6, iNOS and CD86 mRNA, was significantly increased in the of Bv-2 cells after oxygen-glucose deprivation/reoxygenation (OGD/R) model was constructed in vitro. For further understanding the expression pattern of RNAs, the next-generation RNA sequencing was performed and upregulation of Robo4 (roundabout guidance receptor 4) was found both in M1-polarized and OGD/R treated Bv-2 cells, which was also confirmed by RT-qPCR. Extracellular soluble Robo4 (sRobo4) protein also increased in the supernatant of M1-polarized and OGD/R treated Bv-2 cells. Treating bEND3 cells with the Robo4 recombinant protein, M1-polarized Bv-2 cell supernatant or OGD/R Bv-2 cell supernatant decreased trans-endothelial electrical resistance (TEER), suggesting the injury of BBB. In addition, Robo4 was also highly expressed in the serum of patients who experienced acute ischemia stroke and mechanical thrombectomy operation. All the results suggest that increased secretion of Robo4 by M1-polarized-microglia during cerebral ischemia-reperfusion is most likely one of the causes of BBB injury, and Robo4 may be one of the therapeutic targets for BBB functional protection. 

Key words: cerebral ischemia-reperfusion; blood-brain barrier; microglia; M1 polarization; sRobo4

Received: 　　Accepted:

Corresponding author: 杨志刚　　E-mail: yang.zhigang1@zs-hospital.sh.cn

DOI: 10.13294/j.aps.2022.0052

Citing This Article：

HUANG Jin-Long, LI Chen, YANG Liang-Liang, GAO Yang, ZHAO Pu-Yuan, YANG Zhi-Gang. The destructive role of soluble Robo4 secreted by the M1-polarized-microglia during cerebral ischemia-reperfusion in blood-brain barrier integrity. Acta Physiol Sin 2022; 74 (4): 513-524 (in Chinese with English abstract).