复方青黛饮含药血清对体外HaCaT细胞增殖和凋亡的影响

Effects of Compound Indigo Naturalis Decoction Serum on Proliferation and Apoptosis of HaCaT Cells In Vitro

目的体外观察复方青黛饮大鼠含药血清对人永生化角质形成细胞株HaCaT细胞增殖、凋亡和细胞周期的影响,并探讨其作用机制。方法根据中药血清药理学方法,用SD大鼠制备实验血清,采用MTT法、流式细胞术、免疫荧光Hoechst染色,观察含药血清对细胞生长抑制率、凋亡率、细胞周期分布及细胞形态学的影响。结果体外培养HaCaT细胞24h和48h后,复方青黛饮高、中、低三个剂量不同浓度的含药血清组,对HaCaT细胞的生长均有不同程度的抑制,与空白血清组相比,差异有统计学意义(P<0.05),其中高剂量组抑制作用最明显。复方青黛饮含药血清组细胞凋亡率和G0/G1比例明显增加,与空白血清组相比,差异有统计学意义(P<0.05),在镜下呈现凋亡的特征性形态改变。结论复方青黛饮大鼠含药血清对体外培养的角质形成细胞株HaCaT细胞有生长抑制和诱导凋亡作用,并对细胞周期有一定的影响。

Objective To investigate the mechanism of compound indigo naturalis decoction in treating psoriasis, we observed the effect of inactivated mouse serum containing compound indigo naturalis decoction on the proliferation, apoptosis, and cell cycle of HaCaT cell in vitro. Methods The experimental animal serum was taken from SD rats and was inactivated before the experiment on the base of the serum pharmacology method. Then the serum at different concentration of compound indigo naturalis decoction serum was added to HaCaT cell. The control was set meanwhile. The cell growth was evaluated by methyl thiazolyl tetrazolium （MTT） assay. The cell cycle distribution and cell apoptosis were measured by flow cytometry. The cell morphology was observed by Hoechst staining under fluorescence microscope. Results After co-culture for 24h, 48h, compared with the blank group, the cell proliferation of HaCaT was inhibited significandy in high dose, medium dose, low dose groups （ P 〈 0.05 ）. Compared with that of the blank group, the cell apoptotic rate and cell percentage in G0/G1 phase obviously increased in high dose, medium dose, low dose groups（P 〈 0. 05）. At the mean time, the distinctive apoptotic morphology was observed. Conclusion We concluded that compound indigo naturalis decoction serum not only inhibited the proliferation of HaCaT cells, but also significantly induce apoptosis. Affecting cell cycle might contribute to the mechanism.