德国洋甘菊精油对脂多糖诱导的人角质形成细胞炎性反应的调控作用研究

Study on the Regulatory Effect of Essential Oil in German Chamomile on the Inflammatory Response of HaCaT Cells Induced by LPS

目的研究德国洋甘菊精油(MCEO)对脂多糖(LPS)诱导的人角质形成细胞(HaCaT细胞)炎症损伤的保护作用及作用机制。方法德国洋甘菊精油预处理HaCaT细胞24 h后,用CCK-8和Western Blot法确定德国洋甘菊精油最佳使用浓度为50μg·mL^(-1),以此进行后续实验。实验分为3组:空白组、模型组和精油处理组(50μg·mL^(-1)预处理24 h)。除空白组外,其余组以10μg·mL^(-1)的脂多糖刺激1 h进行模型复制。通过qRTPCR检测炎性细胞因子IL-6和IL-1β表达水平;Western Blot和免疫荧光法观察丝裂原活化蛋白激酶p38(p38 MAPK)和哺乳动物雷帕霉素靶蛋白(mTOR)信号通路蛋白表达水平。结果与空白组比较,模型组HaCaT细胞中IL-6和IL-1βmRNA表达增加(P<0.0001),p-p38 MAPK和p-mTOR蛋白表达水平明显升高(P<0.05,P<0.001);与模型组比较,精油处理组明显下调了脂多糖诱导的HaCaT炎性细胞因子IL-6 mRNA表达(P<0.01)及p-p38 MAPK和p-mTOR蛋白表达水平(P<0.05,P<0.0001)。结论德国洋甘菊精油可通过p38 MAPK和mTOR两条信号通路共同调控来降低脂多糖诱导的炎性损伤,为银屑病或其他炎症性皮肤病的预防和治疗提供新的思路。

Objective To study the protective effect and mechanism of essential oil in German chamomile(MCEO)on lipopolysaccharide(LPS)-induced inflammatory injury of human keratinocytes(HaCaT)cells.Methods CCK-8 and Western Blot were used to confirm the optimal concentration of MCEO(50μg·mL^(-1))after pretreatment of HaCaT cells with MCEO for 24 h.The experiment was divided into 3 groups:blank group,model group and essential oil group(pretreated with 50μg·mL^(-1) of MCEO for 24 h).Except the blank group,the other groups were stimulated with 10μg·mL^(-1) of LPS for 1 h.The expression levels of inflammatory cytokines IL-6 and IL-1βwere detected by qRT-PCR.The protein expressions of p38 MAPK and mTOR signaling pathway were observed by Western Blot and immunofluorescence assay.Results Compared with the blank group,the mRNA expressions of IL-6 and IL-1βin HaCaT cells in the model group were increased(P<0.0001),and the protein expressions of p-p38 MAPK and pmTOR were significantly increased(P<0.05,P<0.001).Compared with the model group,MCEO significantly down-regulated LPS-induced mRNA expression of IL-6(P<0.01)and protein expressions of p-p38 MAPK and pmTOR(P<0.05,P<0.0001)in HaCaT.Conclusion MCEO can reduce LPS-induced inflammatory injury through the co-regulation of p38 MAPK and mTOR signaling pathways.This experiment is expected to provide new ideas for the prevention and treatment of psoriasis or other inflammatory skin diseases.