Action mechanism by which fibronectin type III domain-containing protein 5 inhibits macrophage pyroptosis

doi:10.12307/2024.196

Abstract

Abstract: BACKGROUND: Fibronectin type III domain-containing protein 5 (FNDC5) is a muscle factor that can regulate glucose and lipid metabolism, and exert anti-inflammatory, anti-oxidative effects and improvement in insulin resistance. Moreover, FNDC5 could control various cell pyroptosis.
OBJECTIVE: To explore the effect and potential mechanism of FNDC5 on macrophage pyroptosis.
METHODS: (1) After completing the construction of the lentivirus virus overexpressing FNDC5 or silencing FNDC5, the THP-1 cells were transfected with the lentivirus vector. The result of transfection was detected by the expression of green fluorescence, qPCR, and western blot assay. (2) Phorbol ester induced THP-1 cells to differentiate into macrophages. Oxidized low-density lipoprotein (ox-LDL) was added to induce the cell pyroptosis model. There were six groups, i.e., NC group, ox-LDL group, ox-LDL+MOCK1 group, ox-LDL+Ov-FNDC5 group, ox-LDL+MOCK2 group, and ox-LDL+shFNDC5 group. (3) The level of cell pyroptosis was evaluated by Hoechst 33342/propidium iodide fluorescence double staining and lactate dehydrogenase release assay. The expression levels of related molecules in THP-1 cells were analyzed by qPCR and western blot assay. The interleukin-18 and interleukin-1β in cell supernatant were detected by ELISA.
RESULTS AND CONCLUSION: Compared with the ox-LDL+MOCK1 group, overexpression of FNDC5 significantly reduced the pyroptosis rate of macrophages and the release levels of lactate dehydrogenase, interleukin-1β and interleukin-18, significantly inhibited the mRNA expression of NF-κB p65, NF-κB p50, NLRP3, ASC, Caspase-1, and GSDMD, and significantly inhibited the protein expression of NF-κB p65, NF-κB p50, NLRP3, ASC, cleaved Caspase-1 and GSDMD-N. Compared with the ox-LDL+MOCK2 group, the silence of FNDC5 showed the opposite result. These findings suggest that FNDC5 attenuates pyroptosis in macrophages by inhibiting the NF-κB/NLRP3 pathway.

Key words: macrophage, fibronectin type III domain-containing protein 5, FNDC5, NF-κB, NLRP3, cell pyroptosis

CLC Number:

Zhao Guangjian, Liu Danan, Zhou Bo, Wang Yao. Action mechanism by which fibronectin type III domain-containing protein 5 inhibits macrophage pyroptosis[J]. Chinese Journal of Tissue Engineering Research, 2024, 28(25): 4005-4012.

Figures/Tables 12

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