Chemokine C-C motif ligand 2 suppressed the growth of human brain astrocytes under Ischemic/hypoxic conditions via regulating ERK1/2 pathway

ABSTRACT

Primary Objective: Chemokine C-C motif ligand 2 (CCL2) plays a critical role in inflammation-related diseases in the central nervous system (CNS). However, the role of CCL2 in ischemic stroke remains unclear. Research Design: To investigate the role of CCL2 in ischemic stroke, we performed oxygen–glucose deprivation (OGD) on human brain astrocytes. Methods and Procedures: To assess cell proliferation, the CCK-8 assay was performed. Cell apoptosis was determined using flow cytometry. qRT-PCR and western blotting were utilized to measure gene expression. Main Outcomes and Results: Our results suggest that CCL2 and its receptor CCR2 are upregulated in OGD cells. Moreover, a CCL2 antibody significantly alleviated the ischemic/hypoxic-induced suppression of growth in human brain astrocytes. Human recombinant protein, CCL2, inhibited the growth of human brain astrocytes under normoxia conditions. These results demonstrate that CCL2 upregulation suppresses the recovery of human brain astrocytes under ischemic/hypoxic conditions. This effect was abolished by the ERK inhibitor PD98059. Therefore, CCL2/CCR2 activation may suppress the growth of human brain astrocytes through enhancing the activity of ERK1/2. Conclusions: Our results not only developed a deeper understanding of the role of CCL2 in human brain astrocytes but also provided novel insight into potential treatments for ischemic stroke.

KEYWORDS:

• Ischemic stroke
• astrocytes
• CCL2/CCR2
• ERK1/2

Acknowledgments

We sincerely acknowledge the assistance given by the Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine, Shanghai, China, and Shanghai 9th People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China. Qing Zhan and Jiangmin Zhao designed this project and wrote the manuscript; Min Yu and Ni Zheng performed the experiments; Dudu Jiang and Lijing Wang analyzed the data and edited diagrams. All authors have contributed to, read, and agreed upon the final contents of the manuscript for submission. The authors would like to thank Enago (www.enago.cn) for the English language review.

Disclosure of interest

The authors report no conflict of interest.

Supplementary material

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Additional information

Funding

The study was supported by grants from Key Specialty Construction Project of Pudong Health and Family Planning Commission of Shanghai [No. PWZxK2017-04] and Talents Training Program of Seventh People’s Hospital of Shanghai University of Traditional Chinese Medicine [No. QMX2017-03 and No. MZY2017-01].