龙胆泻肝汤对人角质形成细胞株HaCaT细胞增殖的影响

摘要

Objective:To determine the effect of Longdanxiegan recipe on the proliferation of HaCaT cells. Methods:HaCaT cells were cultured in vitro,and then, were divided into Longdanxiegan recipe group with different concentrations(20 μg/mL,40 μg/mL,60 μg/mL,80 μg/mL and 100 μg/mL) and control group. The cell cycle was determined by FACS,and the level of DNA replication was analyzed by BrdU incorporation assay,The level of proliferation was analyzed by MTT assays and the levels of CDK4/Cyclin D1and phospho-rylated RB protein were detected by realtime PCR and Western blot. Results:Compared to the control group, in the Longdanxiegan recipe group,the ratio of G1 cells was higher,but OD score,BrdU positive cells,and the levels of p-RB and CDK4/Cyclin D was lower.Conclusion:Longdanxiegan recipe can inhibit the prolifer-ation of HaCaT cells and the mechanism may be associated with the down-regulation of CDK4/Cyclin D1.%目的: 确定龙胆泻肝汤对人角质形成细胞株HaCaT细胞增殖的影响.方法: 体外培养HaCaT细胞,分为龙胆泻肝汤处理组(20 μg/mL,40 μg/mL,60 μg/mL,80 μg/mL和100 μg/mL)和空白对照组,使用流式细胞技术检测细胞周期分布,BrdU掺入法检测DNA复制水平,MTT法检测细胞增殖情况,Realtime PCR和Western blot检测关键增殖调控蛋白(CDK4/Cyclin D1和磷酸化Rb蛋白)的表达.结果: 与对照组比较,龙胆泻肝汤处理组 G1期细胞比例升高,OD 值、BrdU 阳性细胞比例及CDK4/Cyclin D1、磷酸化Rb蛋白表达下降.结论: 龙胆泻肝汤抑制HaCaT细胞增殖,其机制可能与抑制CDK4/Cyclin D1活性有关.