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作 者:李欣[1,2] 陈再明 葛维维 吕曹华[3] 陈光[2,3] 张雪松 LI Xin;CHEN Zaiming;GE Weiwei;LYU Caohua;CHEN Guang;ZHANG Xuesong(Jiamusi University,Jiamusi 154000 Heilongjiang,China;Medical College of Taizhou University,Taizhou 318000 Zhejiang,China;Taizhou Second People’s Hospital,Taizhou 318000 Zhejiang,China)
机构地区:[1]佳木斯大学研究生学院,黑龙江佳木斯154000 [2]台州学院医学院,浙江台州318000 [3]台州市第二人民医院,浙江台州318000
出 处:《中药新药与临床药理》2022年第5期624-632,共9页Traditional Chinese Drug Research and Clinical Pharmacology
基 金:台州市科技计划项目(20ywa57)。
摘 要: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.
关 键 词:德国洋甘菊精油 银屑病 人角质形成细胞(HaCaT细胞) 脂多糖 炎症 丝裂原活化蛋白激酶p38(p38 MAPK) 哺乳动物雷帕霉素靶蛋白(mTOR)
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