CIRC_0044235 targeting miR-574-5p attenuates IL-1β-induced injury in human chondrocyte

doi:10.16352/j.issn.1001-6325.2023.03.419

Abstract

Abstract: Objective To investigate whether CIRC_0044235 targeting miR-574-5p could affect the levels of chondro-cyte apoptosis, oxidative stress and inflammatory factor expression induced by interleukin (IL)-1β. Methods The cells were divided into control group, IL-1β group, transfected with pcDNA, pcDNA-CIRC_0044235, anti-miR-NC, anti-miR-574-5p, pcDNA-CIRC_0044235+miR-NC, pcDNA-CIRC_0044235+miR-574-5p, interfere with IL-1β. RT-qPCR was used to detect the expression of CIRC_0044235 and miR-574-5p in cell. Flow cytometry and Western blot were used to detect apoptosis and the expression of related proteins Bcl-2 and Bax, respectively. Colorimetry was used to superoxide dismutase (SOD) and lactate dehydrogenase (LDH) activity were detected, and ELISA was used to detect the levels of IL-6 and IL-10. A dual-luciferase reporter assay was used for examining the targeted binding of CIRC_0044235and miR-574-5p.Results Compared with the control group, the expression of CIRC_0044235, the expression of Bcl-2 protein, the activity of SOD and the content of IL-10 in chondrocytes in the IL-1β group were decreased (P＜0.05), while the expression of miR-574-5p and the rate of apoptosis were decreased (P＜0.05). Bax protein expression, LDH activity and IL-6 content all increased(P＜0.05). Compared with IL-1β group or pcDNA intervention group, CIRC_0044235 expression, Bcl-2 protein expression, SOD activity and IL-10 content in chondrocytes of pcDNA-CIRC_0044235 group increased, while miR-574-5p expression, apoptosis rate, Bax protein expression, LDH activity and IL-6 content decreased (P＜0.05). CIRC_0044235 targeted miR-574-5p. miR-574-5p expression, apoptosis rate, Bax protein expression, LDH activity and IL-6 content in chondrocytes of anti-miR-574-5p group were all lower than those found in anti-miR-NC group. Bcl-2 protein expression, SOD activity and IL-10 content were higher than those in anti-miR-NC group (P＜0.05). miR-574-5p expression, apoptosis rate, Bax protein expression, LDH activity and IL-6 content in chondrocytes of pcDNA-CIRC_0044235+miR-574-5p group were higher than those of pcDNA-CIRC_0044235+miR-NC group. The Bcl-2 protein expression, SOD activity and IL-10 content were lower than those in the pcDNA-CIRC_0044235+miR-NC group (P＜0.05).Conclusions CIRC_0044235 may inhibit IL-1β-induced apoptosis, oxidative stress and inflammatory factor expression in human articular chondrocytes by targeting miR-574-5p.

Key words: osteoarthritis, CIRC_0044235, miR-574-5p, apoptosis, oxidative stress

CLC Number:

R684.3

WANG Bo, ZHANG Minghuan. CIRC_0044235 targeting miR-574-5p attenuates IL-1β-induced injury in human chondrocyte[J]. Basic & Clinical Medicine, 2023, 43(3): 419-426.

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