黄连素对人骨关节炎软骨细胞增殖、凋亡及炎性因子分泌的影响

Effect of berberine on the proliferation,apoptosis and inflammatory factor secretion of human chondrocytes osteoarthritis

目的观察黄连素(BER)对人骨关节炎软骨细胞(HC-OA)增殖、凋亡及炎性因子分泌的影响。方法选取HC-OA细胞进行培养,空白对照组进行常规培养,实验组在此基础上加入不同浓度BER进行处理。采用CCK-8试剂盒检测细胞增殖能力变化;实时定量PCR检测凋亡基因BCL-2、BAX、Caspase 3表达情况;应用酶联免疫吸附法(ELISA)检测炎症因子白细胞介素-1β(IL-1β)和肿瘤坏死因子-α(TNF-α)水平。结果BER处理HC-OA细胞72 h后,相比空白对照组,不同浓度的BER均能显著促进HC-OA增殖,且浓度越高促增殖活性越显著(P<0.05)。相比空白对照组,BER能显著下调细胞凋亡基因BAX、Caspase 3表达,上调抗凋亡基因BCL-2表达(P<0.05)。相比空白对照组,经LPS诱导后细胞分泌IL-1β、TNF-α水平显著上调;相比LPS组,LPS+BER组细胞分泌IL-1β、TNF-α水平显著下调(P<0.05)。结论BER能在体外显著干预凋亡基因和抗凋亡基因表达,促进HC-OA增殖、抑制其凋亡,减少炎性因子分泌。

Objective To observe the effect of berberine(BER)on the proliferation,apoptosis and inflammatory factor secretion of human chondrocytes osteoarthritis(HC-OA).Methods HC-OA cells were selected for culture,the blank control group was routinely cultured,and the experimental group was treated with different concentrations of BER on this basis.CCK-8 kit was used to detect the change of cell proliferation ability;real-time quantitative PCR was used to detect the expression of apoptosis genes of BCL-2,BAX and Caspase 3;enzyme linked immunosorbent assay(ELISA)was used to detect the inflammatory factors of interleukin-1β(IL-1β)and tumor necrosis factor-α(TNF-α)levels.Results After HC-OA cells were treated with BER for 72 h,compared with the blank control group,different concentrations of BER could significantly promote the proliferation of HC-OA,and the higher the concentration,the more significant the proliferation activity(P<0.05).Compared with the blank control group,BER could significantly down-regulate the expression of apoptosis genes of BAX and Caspase 3,and up-regulate the expression of anti-apoptosis gene of BCL-2(P<0.05).Compared with the blank control group,the levels of IL-1βand TNF-αsecreted by cells were significantly up-regulated after LPS induction;compared with LPS group,the levels of IL-1βand TNF-αsecreted by cells in the LPS+BER group were significantly down-regulated(P<0.05).Conclusion BER can significantly interfere with the expression of apoptotic genes and anti-apoptotic genes in vitro,promote the proliferation of HC-OA,inhibit its apoptosis and reduce the secretion of inflammatory factors.