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目的观察中药清热活血解毒复方对人永生化角质形成细胞株(HaCaT)增殖、凋亡和细胞周期的影响,探讨其治疗银屑病可能的作用机制。方法将不同浓度的清热活血解毒复方水提物分别作用于体外培养的HaCaT细胞,采用CCK-8法检测药物对细胞增殖的抑制作用及药物的有效浓度;采用倒置显微镜观察药物处理前后细胞的形态学改变;流式细胞术检测细胞周期的变化及凋亡率。结果清热活血解毒复方以时间和浓度依赖性方式抑制HaCaT细胞增殖,作用24、48 h其半抑制浓度(IC50)分别为23.40、20.24 mg/m L。细胞形态学和流式细胞仪检测发现药物以浓度依赖性方式干扰细胞周期,诱导细胞凋亡,细胞分裂周期阻滞于G0/G1期。结论清热活血解毒复方可能通过抑制HaCaT细胞增殖、诱导凋亡、影响细胞周期而发挥治疗银屑病的作用。
Objective To observe the effects of traditional Chinese medicine Qingre Huoxue Jiedu compound on proliferation, apoptosis and cell cycle of human immortalized keratinocyte (HaCaT) cells and to explore its possible mechanism of action in treating psoriasis. Methods Different concentrations of water extracts from heat-clearing, blood-activating and detoxifying treatment were applied to cultured HaCaT cells respectively. The inhibitory effect of drugs on cell proliferation and the effective concentration of drugs were detected by CCK-8 assay. The morphological changes of cells were observed with inverted microscope Learning changes; flow cytometry to detect changes in cell cycle and apoptosis rate. Results The heat-clearing and blood-activating and detoxifying compound inhibited the proliferation of HaCaT cells in a time-and-concentration-dependent manner. The half-inhibitory concentrations (IC50) at 24 and 48 h were 23.40 and 20.24 mg / m L, respectively. Cytometry and flow cytometry showed that the drug interfered with the cell cycle in a concentration-dependent manner, induced apoptosis, and the cell division cycle was arrested in G0 / G1 phase. Conclusion Qingre Huoxue Jiedu compound may play a role in the treatment of psoriasis by inhibiting the proliferation of HaCaT cells, inducing apoptosis and affecting the cell cycle.