Regulatory effects of sphingosine kinase-2 on astrocyte function and EAE progression in mouse model
10.3760/cma.j.cn112309-20200213-00061
- VernacularTitle:鞘氨醇激酶-2对星形胶质细胞功能及实验性自身免疫性脑脊髓炎小鼠疾病进程的影响
- Author:
Jingjing GUO
1
;
Ying YANG
;
Qianwen CAO
;
Zijun ZHAO
;
Xiangyang LI
;
Hui HUA
;
Xiaocui LI
;
Wanpeng CHENG
;
Feng ZHOU
;
Xiaomei LIU
Author Information
1. 徐州医科大学病原生物学与免疫学教研室，江苏省免疫与代谢重点实验室 221004
- From:
Chinese Journal of Microbiology and Immunology
2020;40(10):780-786
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To investigate the regulatory effects of sphingosine kinase-2 (SphK2) on the function of activated astrocytes and the progression of experimental autoimmune encephalomyelitis (EAE) in mice.Methods:Primary mouse astrocytes were isolated from wild-type (WT) C57BL/6 mice and sphk2 gene knock-out ( sphk2 -/-) mice and stimulated in vitro with interleukin 17 (IL-17). Real-time PCR was used to measure the expression of inflammatory cytokines and chemokines at mRNA levels. Western blot and immunofluorescence were used to detect the expression of glial fibrillary acidic protein (GFAP) and the phosphorylation of signal transducer and activator of transcription 3 (p-STAT3). An EAE mouse model was constructed using myelin oligodendrocyte glycoprotein 35-55 (MOG 35-55) polypeptide. Western blot was used to detect the expression of GFAP and p-STAT3 at protein level and real-time PCR was used to detect the expression of inflammatory cytokines and chemokines at mRNA level in spinal cords. Hematoxylin-Eosin (HE) and Luxol fast blue (LFB) staining were used to observe the changes in inflammatory cell infiltration and demyelination in spinal cords. Results:Compared with the WT group, the phosphorylation of STAT3 was obviously reduced in in vitro activated mouse astrocytes of sphk2 -/- mice, and the expression of inflammatory cytokines and chemokines including monocyte chemotactic protein 1 (MCP-1), TNF-α and IL-6 at mRNA level was also significantly decreased. Compared with the WT EAE group, changes in the above-mentioned cytokines and relative proteins in sphk2 -/- EAE mice in vivo were similar to those in vitro. Moreover, inflammatory cell infiltration and demyelination were significantly reduced in spinal cords of sphk2 -/- EAE mice. However, no significant difference in in vitro or in vivo GFAP expression was observed between WT and sphk2 -/- mice. Conclusions:SphK2 might regulate the function of reactive astrocytes through STAT3 molecular pathway, thereby regulating the production of inflammatory cytokines and chemokines and participating in the pathological process of EAE.
