积雪草酸对TNF-α诱导的HaCaT细胞增殖的抑制作用

Inhibitory effect of asiatic acid on the HaCaT cell proliferation induced by TNF-α

目的探讨积雪草酸对TNF-α诱导的人角化细胞系HaCaT增殖的影响及其可能机制。方法分别通过MTT实验和PI染色流式细胞术检测不同浓度TNF-α对体外HaCaT细胞增殖能力和细胞周期的影响。采用免疫荧光染色法检测不同浓度积雪草酸对TNF-α诱导HaCaT细胞NF-κB核转位的影响。通过Western印迹法检测NF-κB信号通路中p-NF-κB、NF-κB的表达水平。结果 TNF-α诱导的HaCaT增殖实验结果显示,与正常对照组相比,模型组中细胞存活率明显升高(P<0.01),S期细胞比例显著升高(P<0.05),G2期细胞比例显著降低(P<0.05),p-NF-κB表达显著升高(P<0.01);与模型组相比,10、25和50μmol/L积雪草酸组细胞存活率显著下降(均为P<0.05),10和40μmol/L积雪草酸组G2期细胞比例显著升高(P<0.05),pNF-κB表达显著降低(P<0.05)。结论积雪草酸可能通过抑制NF-κB信号通路来抑制角质形成细胞的增殖,可以作为治疗银屑病的潜在药物。

Objective To investigate the effect of asiatic acid on the keratinocytes HaCaT cell proliferation induced by tumor necrosis factor-α(TNF-α)in vitro and its possible mechanism. Methods The effects of TNF-α on the proliferation and cell cycle of HaCaT cells in vitro were detected by the MTT assay and PI staining flow cytometry,respectively. The effect of different concentrations of asiatic acid on NF-κB nuclear translocation in HaCaT cells was determined by immunofluorescence staining,and Western blotting was carried out to determine the expression level of p-NF-κB and NF-κB both in the NF-κB signaling pathway. Results In the model test,compared with the normal control group,the cell viability(P<0.05),proportion of the S phase cells(P<0.05)and the p-NF-κB expression(P<0.01)were significantly increased,whereas the proportion of cells was significantly decreased(P<0.05). Compared with the model group,the cell viability and the p-NF-κB expression were significantly decreased in the 10,25 and 50 μmol/L asiatic acid group(both P<0.05),while the proportion of G2 phase cells in the 10 and 40 μmol/L asiatic acid group was significantly increased(P<0.05). Conclusion Asiatic acid could inhibit the proliferation of keratinocytes by inhibiting the NF-κB signaling pathway and may be a potential candidate for the treatment of psoriasis.