双氢青蒿素和氧化苦参碱对HaCaT细胞增殖及细胞因子表达的影响

Effects of dihydroartemisinin and oxymatrine on the proliferation and cytokine expression of HaCaT cells

[目的]探讨双氢青蒿素和氧化苦参碱对人角质形成细胞HaCaT细胞增殖及细胞因子表达的影响.[方法]用0,10,20,30,40 mg/L质量浓度双氢青蒿素和0,50,100,200,400 mg/L质量浓度的氧化苦参碱处理HaCaT细胞,用MTT法检测HaCaT细胞活力,RT-PCR法检测HaCaT细胞炎症相关细胞因子的表达水平,ELISA法检测细胞培养上清液中白细胞介素(IL)-17A和IL-23的表达水平.[结果]双氢青蒿素明显抑制HaCaT细胞增殖(P<0.05),且具有时间和浓度依赖性,而氧化苦参碱对HaCaT细胞增殖未见有明显抑制作用.双氢青蒿素和氧化苦参碱均显著增加HaCaT细胞抑炎性细胞因子IL-4和IL-10的表达(P<0.05),显著降低促炎性细胞因子人干扰素-γ,IL-17,IL-23的表达(P<0.05),同时氧化苦参碱显著增加抑炎性细胞因子IL-6的表达(P<0.05).ELISA法检测结果显示,双氢青蒿素和氧化苦参碱均明显降低细胞培养上清中IL-17A和IL-23的表达水平(P<0.05).[结论]双氢青蒿素和氧化苦参碱可能通过IL-23/IL-17轴影响HaCaT细胞炎性细胞因子的表达,认为有望成为治疗银屑病的新的中药单体.

OBJECTIVE To investigate the effects of dihydroartemisinin and oxymatrine on the proliferation and cytokine expression of human keratinocyte HaCaT cells. METHODS HaCaT cells were treated with dihydroartemisinin at mass concentrations of 0, 10, 20, 30, 40 mg/L and oxymatrine at mass concentrations of 0, 50, 100, 200, 400 mg/L. The activity of HaCaT cells was detected by MTT assay. RT-PCR was used to detect the expressions of inflammatory-related cytokines in HaCaT cells, and ELISA was used to detect the levels of IL-17 A and IL-23 in cell culture supernatant. RESULTS Dihydroartemisinin significantly inhibited the proliferation of HaCaT cells in time and concentration dependent manner(P< 0.05), while oxymatrine did not significantly inhibit the proliferation of HaCaT cells. Both dihydroartemisinin and oxymatrine significantly increased the expressions of anti-inflammatory cytokines IL-4 and IL-10(P< 0.05), and significantly decreased the expressions of pro-inflammatory cytokines IFN-γ, IL-17, IL-23(P< 0.05). While oxymatrine significantly increased the expression of anti-inflammatory cytokine IL-6(P< 0.05). ELISA results showed that both dihydroartemisinin and oxymatrine obviously reduced the expression levels of IL-17 A and IL-23 in cell culture supernatant(P< 0.05). CONCLUSION Dihydroartemisinin and oxymatrine might affect the expression of inflammatory cytokines in HaCaT cells through the IL-23/IL-17 axis, they were expected to become new monomers of traditional Chinese medicine in the treatment of psoriasis.