Piezo 1 mediates apoptosis of fibroblast-like synovial cells in rheumatoid arthritis via MAPK/ERK5 signaling pathway

doi:10.12307/2021.114

Abstract

Abstract:

BACKGROUND: Previous studies have mostly focused on immune factors involved in the pathogenesis of rheumatoid arthritis. Recently, increasing attention has been paid to the role of biomechanical factors in the occurrence and development of rheumatoid arthritis.
OBJECTIVE: To explore the mechanism by which Piezo1 mediates apoptosis of fibroblast-like synovial cells in rheumatoid arthritis through MAPK/ERK5 signaling pathway.
METHODS: The tissue block method was used to culture synovial cells of rheumatoid arthritis. The stretch stress model of cells in vitro was constructed by Flexcell 5000T cell stretch stress system, and piezo1 siRNA gene interference vector and overexpression plasmid were constructed. According to the preliminary experimental results and treatment plan, fibroblast-like synovial cells of rheumatoid arthritis were divided into six groups: siRNA interference group, overexpression plasmid group, blank control group, siRNA interference+BIX02188 group, overexpression plasmid+BIX02188 group, and blank control+BIX02188 group. The expression of Piezo1, ERK5 and apoptosis-associated genes were detected by RT-PCR, the intracellular calcium content was detected by Fluo-3 AM probe, and the apoptotic level was detected by AV-PI kit.
RESULTS AND CONCLUSION: The intracellular Ca2+ content of the siRNA interference+BIX02188 group was significantly lower than that of the siRNA interference group (P < 0.05); the intracellular Ca2+content of the overexpression plasmid+BIX02188 group was significantly lower than that of the overexpression plasmid group (P < 0.05). The relative expression of Piezo1 and ERK5 mRNA in the siRNA interference group was significantly lower than that in the blank control group (P < 0.05), and that in the overexpression plasmid group was significantly higher than that in the blank control group (P < 0.05). The expression of Piezo1 mRNA did not change after BIX02188 inhibition; however, the relative expression of ERK5 mRNA in the siRNA interference+BIX02188 group was significantly lower than that in the siRNA interference group (P < 0.05), and the expression of ERK5 mRNA in the overexpression plasmid+BIX02188 group was significantly lower than that in the overexpression plasmid group (P < 0.05). The early apoptotic rate, the late apoptotic rate and the total apoptotic rate in the siRNA interference group were significantly lower than those in the blank control group (P < 0.05), whereas the early apoptotic rate, the late apoptotic rate and the total apoptotic rate in the overexpression plasmid group were significantly higher than those in the blank control group (P < 0.05). To conclude, the overexpression of Piezo 1 protein can promote the apoptosis of rheumatoid arthritis fibroblast-like synovial cells, and the apoptotic signal mediated by the MAPK/ERK5 signaling pathway can act as the potential target gene for treating rheumatoid arthritis.

Key words: synovial cells, rheumatoid arthritis, mechanical sensitive ion channels, apoptosis, calcium ion, pathway, stretch stress

CLC Number:

Qu Xiangyang, Song Qinyong. Piezo 1 mediates apoptosis of fibroblast-like synovial cells in rheumatoid arthritis via MAPK/ERK5 signaling pathway[J]. Chinese Journal of Tissue Engineering Research, 2021, 25(26): 4156-4161.

Figures/Tables 7

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