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目的研究小檗碱衍生物HB-13对人永生化角质形成细胞(HaCaT细胞)分泌γ干扰素(IFN-γ)的影响,探讨HB-13的抗炎机制。方法采用四甲基偶氮唑蓝法确定HB-13作用于HaCaT细胞的安全浓度;细胞经P物质刺激后,加入不同浓度的HB-13分别孵育12、24、48h,应用酶联免疫吸附法(ELISA)检测培养上清液中IFN-γ的含量,应用实时荧光定量PCR法检测细胞内IFN-γmRNA表达情况。结果 HB-13浓度为2.5、5、10μg/ml对HaCaT细胞毒性作用不显著,并可促进P物质诱导的IFN-γ的分泌。结论促进表皮细胞分泌IFN-γ可能是HB-13在皮肤局部发挥抗炎作用的机制之一。
Objective To investigate the effects of HB-13, a derivative of berberine, on the secretion of IFN-γ by human immortalized keratinocytes (HaCaT cells) and the anti-inflammatory mechanism of HB-13. Methods The safe concentration of HB-13 on HaCaT cells was determined by MTT method. After the cells were stimulated with substance P, different concentrations of HB-13 were incubated for 12, 24 and 48 hours, respectively. Enzyme-linked immunosorbent assay (ELISA) was used to detect the content of IFN-γ in culture supernatant. The expression of IFN-γmRNA was detected by real-time fluorescence quantitative PCR. Results The cytotoxicity of HB-13 at concentrations of 2.5, 5 and 10 μg / ml on HaCaT cells was not significant, and the secretion of IFN-γ induced by substance P was promoted. Conclusions Promoting the secretion of IFN-γ by epidermal cells may be one of the mechanisms by which HB-13 exerts an anti-inflammatory effect on the skin.