Astragaloside inhibits astrocyte activation and inflammatory response induced by inflammation

doi:10.12307/2024.726

Chinese Journal of Tissue Engineering Research ›› 2024, Vol. 28 ›› Issue (31): 5022-5028.doi: 10.12307/2024.726

Astragaloside inhibits astrocyte activation and inflammatory response induced by inflammation

Yu Jingwen1, Guo Minfang1, Zhang Bingxin1, Mu Bingtao1, Meng Tao1, Zhang Huiyu1, Ma Cungen1, 2, Yin Jinzhu3, Song Lijuan2, 3, Yu Jiezhong1, 2, 4

1Institute of Brain Science/Key Laboratory of Molecular Cellular Immunology in Datong City/Department of Neurology of First Affiliated Hospital, Shanxi Datong University, Datong 037009, Shanxi Province, China; 2Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China;

Received:2023-09-06 Accepted:2023-11-01 Online:2024-11-08 Published:2024-01-22
Contact: Yu Jiezhong, MD, Professor, Institute of Brain Science/Key Laboratory of Molecular Cellular Immunology in Datong City/Department of Neurology of First Affiliated Hospital, Shanxi Datong University, Datong 037009, Shanxi Province, China; Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; The Fifth People’s Hospital of Datong, Datong 037009, Shanxi Province, China Song Lijuan, MD, Associate professor, Key Research Laboratory of Benefiting Qi for Acting Blood Circulation Method to Treat Multiple Sclerosis of State Administration of Traditional Chinese Medicine/Research Center of Neurobiology, Shanxi University of Chinese Medicine, Jinzhong 030619, Shanxi Province, China; Department of Neurosurgery, Tongmei General Hospital/Key Laboratory of Prevention and Treatment of Neurological Diseases, Shanxi Provincial Health Commission, Datong 037003, Shanxi Province, China
About author:Yu Jingwen, Master, Senior experimentalist, Institute of Brain Science/Key Laboratory of Molecular Cellular Immunology in Datong City/Department of Neurology of First Affiliated Hospital, Shanxi Datong University, Datong 037009, Shanxi Province, China
Supported by:
Shanxi Basic Research Program, No. 20210302123337 (to YJW); Shanxi Basic Research Program, No. 20210302123478 (to ZHY); University Level Research Project of Shanxi Datong University, No. 2022K17 (to YJW); Innovation and Entrepreneurship Training Program of Shanxi College Students, No. XDC2022119 (to ZBX); Medical Science and Technology Leading Team of Shanxi Provincial Health Commission, No. 2020TD05 (to MCG); Zhang Zhongjing Inheritance and Innovation Project of National Administration of Traditional Chinese Medicine, No. GDY-KJS-2022-048-1 (to SLJ); 2022 Shanxi Science and Technology Innovation Young Talent Team, No. 202204051001028 (to SLJ); 2022 Traditional Chinese Medicine Research Project Plan of Shanxi Provincial Health Commission, No. 2022ZYYC090 (to MCG); Young Scientist Training Project of Shanxi University of Chinese Medicine, No. 2021-PY-QN-09 (to SLJ); 2022 Annual Science and Technology Innovation Team of Shanxi University of Chinese Medicine, No. 2022TD2010 (to MCG, SLJ)

Abstract

Abstract: BACKGROUND: Astrocytes play an important role in the pathology of central nervous system diseases. The phenotypic and functional changes in astrocytes suggest that it may be an effective therapeutic target for central nervous system diseases. Our previous studies have confirmed that astragaloside can inhibit the lipopolysaccharide-induced astrocyte inflammatory response. Whether astragaloside can regulate the phenotype and function of astrocytes through Notch-1 and its downstream signaling pathway remains unclear.
OBJECTIVE: To explore the effect of astragaloside on astrocyte activation and inflammatory response induced by inflammation and its possible mechanism.
METHODS: Cerebral cortex astrocytes derived from neonatal C57BL/6 mouse were cultured in vitro. CCK-8 assay was used to determine the optimum concentration of astragaloside and Notch active inhibitor DAPT. The astrocytes were divided into five groups: PBS group, lipopolysaccharide group, lipopolysaccharide + astragaloside group, lipopolysaccharide + DAPT group and lipopolysaccharide + DAPT + astragaloside group. The secretion level of inflammatory factors was detected by ELISA, and the level of nitric oxide was detected by Griess method. The astrocytes and splenic mononuclear cells were co-cultured in Transwell chamber to observe the migration of CD4T cells. The expression of astrocyte activation marker GFAP, A1 marker C3 and A2 marker S100A10 as well as Notch 1 and Jag-1 was detected by immunofluorescence staining. The expressions of CFB, C3, S100A10, PTX3, Notch-1, Jag-1, and Hes were detected by western blot assay.
RESULTS AND CONCLUSION: (1) According to the results of CCK8 assay, the final concentration of astragaloside was selected as 25 μmol/L and the final concentration of DAPT was 50 μmol/L for follow-up experiments. (2) Compared with PBS group, interleukin-6, interleukin-12 and nitric oxide secretion levels in the lipopolysaccharide group were significantly increased (P < 0.05, P < 0.05, P < 0.01). Compared with the lipopolysaccharide group, interleukin-6 (all P < 0.05), interleukin-12 (P > 0.05, P < 0.05, P < 0.05) and nitric oxide (P < 0.05, P < 0.01, P < 0.01) secretion significantly reduced in the lipopolysaccharide + astragaloside group, lipopolysaccharide +DAPT group, lipopolysaccharide + DAPT + astragaloside group. (3) Compared with the PBS group, the expression of GFAP that is the marker of activated astrocytes and the migration of CD4 T cells were significantly increased in the lipopolysaccharide group (P < 0.01). Compared with the lipopolysaccharide group, astrocyte activation was significantly inhibited and CD4 T cell migration was significantly reduced in the lipopolysaccharide + astragaloside, lipopolysaccharide +DAPT, lipopolysaccharide + DAPT + astragaloside group (P < 0.05, P < 0.05, P < 0.01). (4) Compared with the PBS group, the expressions of A1 markers C3 and CFB in the lipopolysaccharide group were increased (P < 0.01, P < 0.05), and the expressions of A2 markers S100A10 and PTX3 were decreased (P < 0.01, P < 0.05). Compared with the lipopolysaccharide group, C3 (all P < 0.01) and CFB (both P < 0.05) were significantly reduced and S100A10 (all P < 0.01) and PTX3 (P < 0.05, P < 0.05 and P > 0.05) were increased in the lipopolysaccharide + astragaloside, lipopolysaccharide +DAPT, lipopolysaccharide + DAPT + astragaloside group. (5) Compared with the PBS group, the expressions of Jag-1, Notch-1 and Hes in the lipopolysaccharide group were significantly increased (all P < 0.01). Compared with the lipopolysaccharide group, the expressions of Jag-1 (all P < 0.01), Notch-1 (all P < 0.01) and Hes (P < 0.05, P < 0.01 and P < 0.01) were significantly reduced in the lipopolysaccharide + astragaloside, lipopolysaccharide +DAPT, lipopolysaccharide + DAPT + astragaloside group. (6) The results indicate that astragaloside can promote the transformation of astrocytes from A1 to A2 by regulating Notch-1 signaling pathway, reduce the secretion of inflammatory factors and the migration of CD4 T cells, and thus inhibit astrocyte activation and inflammatory response.

Key words: astragaloside, lipopolysaccharide, astrocyte, Notch-1/Hes signaling pathway, neuroinflammatory response, DAPT

CLC Number:

Yu Jingwen, Guo Minfang, Zhang Bingxin, Mu Bingtao, Meng Tao, Zhang Huiyu, Ma Cungen, Yin Jinzhu, Song Lijuan, Yu Jiezhong. Astragaloside inhibits astrocyte activation and inflammatory response induced by inflammation[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(31): 5022-5028.

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References

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Astragaloside inhibits astrocyte activation and inflammatory response induced by inflammation

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