杯苋甾酮通过上调miR-204表达抑制IL-1β诱导的软骨细胞凋亡的实验研究

doi:10.3969/j.issn.1006-6233.2021.03.004

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Abstract Objective: To investigate the effects of Cyasterone on the expression of microRNA (miR)-204 and the apoptosis of chondrocytes induced by interleukin (IL)-1β. Methods: Human normal chondrocytes C28/I2 were cultured in vitro,and set up ①control group (no treatment),IL-1β group (50 mg/L IL-1β),low-dose Cyasterone group (2.5 mg/L Cyasterone + 50 mg/L IL-1β),medium dose Cyasterone group (5 mg/L Cyasterone + 50 mg/L IL-1β) and high dose Cyasterone group (10 mg/L Cyasterone + 50 mg/L IL-1β); ②normal control group (no transfection), negative control group (transfect NC-siRNA) and miR-204 siRNA group (transfect miR-204 siRNA),three groups were adminstered 50 mg/L IL-1β and 10mg/L Cyasterone. The proliferation of C28/I2 cells was detected by CCK-8 method; the apoptosis of C28/I2 cells was detected by flow cytometry; the levels of IL-6,IL-8 and tumor necrosis factor-α (TNF-α) in the supeRNAtant of C28/I2 cells were detected by enzyme-linked immunosorbent assay (ELISA); the expression of miR-204 in C28/I2 cells was detected by real-time fluorescent quantitative PCR (qRT-PCR); in addition,the protein expressions of Cytochrome C,Bcl-2-related X protein (Bax) and Caspase-3 in C28/I2 cells were detected by Western blot (WB). Results: Compared with those in control group,the cell viability and miR-204 expression level of C28/I2 cells in IL-1β group were significantly lower (P<0.05),and the apoptosis rate,the expression levels of IL-6,IL-8 and TNF-α,the protein expression levels of Cytochrome C,Bax and Caspase-3 were significantly higher (P<0.05); moreover,compared with those in the IL-1β group,with the addition of Cyasterone and the increase of its dosage,the cell viability and the expression level of miR-204 in C28/I2 cells increased gradually (P<0.05),and the apoptosis rate,the expression levels of IL-6,IL-8 and TNF-α,the protein expression levels of Cytochrome C,Bax and Caspase-3 decreased gradually (P<0.05). Compared with negative control group,the apoptosis rate of miR-204 siRNA group were significantly increased (P<0.05). Conclusion: Cyasterone may inhibit the IL-1β-induced apoptosis of chondrocytes by up-regulating the expression of miR-204.

Key words： Cyasterone MicroRNA-204 Interleukin-1β Osteoarthritis Chondrocyte

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http://www.hbyxzzs.cn/EN/10.3969/j.issn.1006-6233.2021.03.004 OR http://www.hbyxzzs.cn/EN/Y2021/V27/I3/368

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