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目的探讨地塞米松对体外培养大鼠肝细胞BRL-3A增殖和凋亡的影响。方法用不同浓度的地塞米松处理BRL-3A细胞,于12、24、48、72、120 h后用MTT检测地塞米松对BRL-3A细胞活性的影响,同时分别用Annexin V-FITC双染法、PI单染色法、实时定量-聚合酶链反应(Real-time PCR)等方法检测地塞米松对BRL-3A细胞周期和凋亡的影响。结果 MTT检测表明,地塞米松能够抑制BRL-3A细胞的活性;Annexin V-FITC双染法分析显示,地塞米松具有显著促进BRL-3A细胞凋亡的效果;PI单染色法检测细胞周期分布情况表明,地塞米松处理后的细胞与对照组相比增殖能力减弱;用Real-time PCR检测促细胞凋亡基因Caspase-3、Caspase-8、Caspase-9和促增殖基因Ccnd1和Jun的mRNA表达情况,结果显示,用地塞米松处理后的细胞中Caspase-3、Caspase-8和Caspase-9均表达上调,而Ccnd1和Jun均表达下调,说明地塞米松能促进BRL-3A细胞的凋亡。结论地塞米松可以促进BRL-3A细胞的凋亡,并抑制其增殖。
Objective To investigate the effects of dexamethasone on the proliferation and apoptosis of cultured rat hepatocytes BRL-3A in vitro. Methods BRL-3A cells were treated with different concentrations of dexamethasone. The effects of dexamethasone on the activity of BRL-3A cells were detected by MTT at 12, 24, 48, 72 and 120 h, and double staining with Annexin V-FITC PI staining and Real-time PCR were used to detect the effect of dexamethasone on cell cycle and apoptosis in BRL-3A cells. Results MTT assay showed that dexamethasone can inhibit the activity of BRL-3A cells; Annexin V-FITC double staining analysis showed that dexamethasone significantly promoted the apoptosis of BRL-3A cells; PI single staining was used to detect cell cycle distribution The results showed that the proliferation of dexamethasone-treated cells was weaker than that of the control group. The mRNAs of the pro-apoptotic genes Caspase-3, Caspase-8, Caspase-9 and Ccnd1 and Jun were detected by Real-time PCR The results showed that the expression of Caspase-3, Caspase-8 and Caspase-9 were upregulated in dexamethasone-treated cells, but both of Ccnd1 and Jun were down-regulated, indicating that dexamethasone can promote the apoptosis of BRL-3A cells . Conclusion Dexamethasone can promote the apoptosis of BRL-3A cells and inhibit its proliferation.