凉血活血复方对体外培养HaCaT细胞增殖和凋亡的影响

Effects of Liangxue Huoxue Complex Prescription on Proliferation and Apoptosis of HaCaT Cells in Vitro

目的观察中药凉血活血复方水醇粗提液对人永生化表皮细胞(HaCaT)的增殖抑制效应和诱导细胞凋亡作用的影响,探讨该复方治疗银屑病的可能机制。方法将不同浓度的凉血活血复方水醇粗提液分别作用于体外培养的HaCaT细胞,采用MTT法检测凉血活血复方水醇粗提液对细胞的增殖抑制作用以及药物的有效浓度;采用倒置显微镜、透射电子显微镜观察细胞处理前后的形态学改变:流式细胞术检测细胞周期的变化及凋亡比率。结果凉血活血复方以时间和浓度依赖性方式抑制HaCaT细胞增殖,作用24、48、72h其IC50分别为155.83 mg/mL、71.57mg/mL,41.27 mg/mL。细胞形态学和流式细胞仪检测发现药物以浓度依赖性的方式干扰细胞周期、诱导细胞凋亡,细胞分裂周期阻皆滞于G2/M期。结论凉血活血复方可能通过抑制角质形成细胞的增殖、诱导其凋亡而治疗银屑病。

Objective To investigate the effects and effective concentration of the ＂traditional Chinese medicine, the crude extract of Liangxue Huoxue complex prescription with water and alcohol, on proliferation and apoptosis of keratinocyte HaCaT cells. Methods Cultured HaCaT cells in vitro were exposed to the crude extract of Liangxue Huoxue complex prescription at different concentrations. Inhibition on cell proliferation was measured by MTT assay. Morphological changes were observed under inverted microscope and transmission electron microscope. Cell cycle and apoptosis were analyzed by flow cytometry. Results The crude extract of Liangxue Huoxue complex prescription inhibited the proliferation of HaCaT cells in a time and concentration-dependent mode. The IC50 of the crude extract of Liangxue Huoxue complex prescription acted to HaCaT cell for 24h, 48h and 72h were 155.83mg/mL, 71.57 mg/mL and 41.27 mg/mL respectively. Cell morphological changes and flow cytometry analysis demonstrated that the crude extract of Liangxue Huoxue complex prescription could arrest HaCaT cells in G2/M phase and induce apoptosis in a concentration-dependent manner. Conclusion The crude extract of Liangxue Huoxue complex prescription might inhibit keratinocytes proliferation and induce apoptosis in the treatment of psoriasis.