广西医科大学学报2020，Vol.37Issue(10)：1757-1765,9.DOI:10.16190/j.cnki.45-1211/r.2020.10.001
miR-455-3p通过靶向BMP2调节地塞米松诱导的软骨细胞自噬性死亡
Effect of miR-455-3p on dexamethasone-induced autophagic death of chondrocytes by targeting BMP2
摘要
Abstract
Objective:To investigate the effect of miR-455-3p on dexamethasone (Dex)-induced autophagic death of chondrocytes by targeting BMP2.Methods: The luciferase reporting assay verified the targeting relationship between miR-455-3p and BMP2.Chondrocytes were isolated from SD rats,and chondrocytes treated with Dex were transfected with miR-455-3p mimic or combined with pcDNA-BMP2 transfection.Chondrocytes were randomly divided into four groups: Control group,Dex group,Dex+mimic group,and Dex+mimic+BMP2 group.The mRNA levels of BMP2,Ki67 and PCNA were detected by RT-PCR.Cell apoptosis was detected by flow cytometry.Cell proliferation was detected by CCK-8 method.LC3 content was detected by immunofiuorescence.Western blotting was used to detect the expression levels of BMP2,Caspase-3,Caspase-9,LC3Ⅱ,LC3Ⅰ,Beclin1 and ATG7 proteins.Results:The results showed that there was a direct targeting relationship between miR-455-3p and BMP2.Compared with the Control group,the rnRNA and protein levels of BMP2 in the Dex group were significantly decreased.The apoptosis rate and the cleaved Caspase-3/Caspase-3,cleaved Caspase-9/Caspase-9 ratios were significantly increased.The cell proliferation multiple was significantly decreased,and the LC3 content was significantly increased.The LC3Ⅱ/LC3Ⅰ ratio,Beclinl and ATG7 protein levels were significantly increased (all P＜0.05).Compared with the Dex group,the mRNA and protein levels of BMP2 in the Dex+mimic group were significantly decreased.The apoptosis rate was significantly increased.The cleaved Caspase-3/Caspase-3 and cleaved Caspase-9/Caspase-9 ratios were significantly increased.The cell proliferation multiple was significantly decreased,and the LC3 content,LC3Ⅱ/LC3Ⅰ ratio,Beclin1 and ATG7 protein levels were significantly increased (all P＜0.05).Compared with the Dex+mimic group,the mRNA and protein levels of BMP2 in the Dex + mimic + BMP2 group were significantly increased.The apoptosis rate,cleaved Caspase-3/Caspase-3 and cleaved Caspase-9/Caspase-9 ratios were significantly decreased.The cell proliferation multiple was significantly increased,and the LC3 content,LC3Ⅱ/LC3Ⅰ ratio,Beclin 1 and ATG7 protein levels were significantly increased (all P＜0.05).Conclusion:MiR-455-3p regulates the autophagic death of chondrocytes induced by Dex by targeting BMP2 and promotes proliferation and autophagy of chondrocytes.关键词
骨关节炎/miR-455-3p/骨形态发生蛋白2/自噬Key words
osteoarthritis/miR-455-3p/bone morphogenetic protein 2/autophagy分类
医药卫生引用本文复制引用
张俊,袁映红,冯志蔚,肖涛,孔亮,蒋科..miR-455-3p通过靶向BMP2调节地塞米松诱导的软骨细胞自噬性死亡[J].广西医科大学学报,2020,37(10):1757-1765,9.基金项目
四川省教育厅科研项目(No.18SXHZ0536)This study was funded by The Education Department of Sichuan Province (No.18SX-HZ0536). （No.18SXHZ0536）
