摘要
目的:探讨苍术提取物对IL-1β诱导的骨关节炎软骨细胞RPOC增殖和凋亡的影响及作用机制。方法:IL-1β诱导RPOC细胞,苍术提取物干预后,四甲基噻唑蓝染色法(MTT)检测细胞增殖,流式细胞仪检测细胞凋亡,Western blot检测CyclinD1、P21、Bcl-2和Bax表达水平,实时荧光定量PCR检测miR-378c表达水平。转染miR-378c抑制剂至RPOC细胞,上述相同方法观察抑制miR-378c表达对IL-1β诱导的RPOC细胞增殖和凋亡的影响。结果:IL-1β可抑制RPOC细胞增殖、上调CyclinD1和Bcl-2蛋白表达,促进RPOC细胞凋亡、P21和Bax蛋白表达以及miR-378c表达。苍术提取物可促进IL-1β诱导的RPOC细胞增殖、上调CyclinD1和Bcl-2蛋白表达,抑制RPOC细胞凋亡、P21和Bax蛋白表达及miR-378c表达。抑制miR-378c表达可促进IL-1β诱导的RPOC细胞增殖、上调CyclinD1和Bcl-2蛋白表达,抑制RPOC细胞凋亡、P21和Bax蛋白表达。过表达miR-378c可逆转苍术提取物对IL-1β诱导的RPOC细胞增殖、凋亡及相关蛋白表达。结论:苍术提取物可促进IL-1β诱导的软骨细胞RPOC增殖,并抑制其凋亡,作用机制与下调细胞中miR-378c表达有关。
Objective:To investigate the effects of Rhizoma Atractylodis extract on the proliferation and apoptosis of IL-1β-induced osteoarthritis chondrocyte RPOC and its mechanism.Methods:After IL-1βinduced RPOC cells were intervened by Rhizoma Atractylodis extract,then cell proliferation was detected by MTT assay,apoptosis was detected by flow cytometry,the expression levels of CyclinD1,P21,Bcl-2 and Bax protein were detected by Western blot,and miR-378c expression level was detected by real-time fluorescent quantitative PCR.The miR-378c inhibitor was transfected into RPOC cells,and the same method described above were used to observe the effects of inhibiting of miR-378c expression on IL-1β-induced RPOC cells proliferation and apoptosis.Results:IL-1βinhibited RPOC cells proliferation and the levels of CyclinD1 and Bcl-2 protein,and promoted RPOC cells apoptosis and the levels of P21 and Bax protein as well as miR-378c.Rhizoma Atractylodis extract promoted IL-1β-induced RPOC cells proliferation and the levels of CyclinD1 and Bcl-2 protein,and inhibited RPOC cells apoptosis and the levels of P21 and Bax protein as well as miR-378c.Inhibition of miR-378c expression promoted IL-1β-induced RPOC cells proliferation and the levels of CyclinD1 and Bcl-2 protein,and inhibited RPOC cells apoptosis and the levels of P21 and Bax protein.Overexpression of miR-378c reversed the effect of Rhizoma Atractylodis extract on IL-1β-induced RPOC cells proliferation,apoptosis and related protein expression.Conclusion:Rhizoma Atractylodis extract can promote the proliferation of IL-1β-induced RPOC cells and inhibit its apoptosis.The mechanism of action is related to the down-regulation of miR-378c expression in cells.
作者
蒲博强
邢晓伟
郭祥(指导)
PU Bo-Qiang;XING Xiao-Wei;GUO Xiang(Department of Orthopedics,Haikou Affiliated Hospital of Central South University Xiangya School of Medicine,Haikou 570208,China)
出处
《中国免疫学杂志》
CAS
CSCD
北大核心
2021年第2期167-173,共7页
Chinese Journal of Immunology
基金
海南省卫生计生行业科研项目(18A200115)。
