摘要
目的蜕皮甾酮(Ecdysterone,EDS)是来源于中药牛膝的一类甾酮类化合物。笔者检测了蜕皮甾酮对肿瘤坏死因子(TNF)-α刺激后HaCaT角质形成细胞的炎症因子产生的影响及相关可能机制。方法采用CCK-8细胞活力测定法检测了蜕皮甾酮对细胞增殖的影响,研究蜕采用酶联免疫吸附试验(ELISA)检测了蜕皮甾酮对TNF-α刺激后HaCaT炎症因子产生的影响;采用Western印迹法对NF-κB通路和MAPK通路的相关蛋白进行了检测;采用免疫荧光法检测了TNF-α刺激后HaCaT细胞核因子(NF)-κB蛋白核转位。结果在所选浓度内,蜕皮甾酮对HaCaT细胞增殖无明显影响;蜕皮甾酮抑制了TNF-α诱导的HaCaT细胞胸腺和活化调节趋化因子(TARC),人巨噬细胞来源的趋化因子(MDC/CCL22),调节激活正常T-细胞表达分泌因子(RANTES/CCL5),白细胞介素(IL)-8的表达。蜕皮甾酮能剂量依赖性降低TNF-α诱导的HaCaT细胞胞核内P65的水平,抑制P65和核转位,提高胞浆内IκB的水平;蜕皮甾酮能剂量依赖性抑制TNF-α诱导的HaCaT细胞MAPK通路中P38,ERK,JNK蛋白的磷酸化。结论蜕皮甾酮可能通过调节NF-κB通路和MAPK通路的活化抑制了TNF-α刺激后HaCaT炎症因子产生。这为蜕皮甾酮应用于炎症性皮肤疾病提供了一定的依据。
Objective Ecdysterone (EDS) is a kind of chemical substance derived from Achyranthes Bidentata. We examine the effects of ecdysterone on the production of inflammatory factors in HaCaT cells induced by tumor necrosis factor (TNF-α) stimulation and investigate the possible mechanisms. Methods CCK-8 assay was used to detect the effects of ecdysterone on HaCaT cells proliferation. Enzyme linked immunosorbent assay (ELISA) was used to detect the effect of ecdysterone on the production of inflammatory factors in HaCaT induced by TNF -α stimulation. Western blotting was used to detect the expression of these proteins involved in NF-κB pathway and MAPK pathway. Meanwhile,the nuclear translocation of NF-κB P65 was detected by immunofluorescence in HaCaT cells induced by TNF -α stimulatetion. Results In the selected concentration,ecdysterone had no significant effects on the proliferation of HaCaT cells. Ecdysterone inhibited the expression of TARC,MDC,RANTES and IL-8 in HaCaT cells induced by TNF-α stimulation. Ecdysterone reduced the level of P65 in cytoplasm,raised the level of IκB in nucleus and inhibited nuclear translocation of P65 in HaCaT cells induced by TNF-α stimulation in a dose-dependent manner;Ecdysterone inhibited the phosphorylation of P38,ERK and JNK proteins in HaCaT cells induced by TNF -α stimulation in a dose -dependent manner. Conclusion Ecdysterone inhibited the production of inflammatory factors in HaCaT cells induced by TNF-α stimulation by regulating the activation of the NF-κB pathway and the MAPK pathway. It provides some evidences for the application of ecdysterone in inflammatory skin diseases.
作者
胡惠清
李静
方坤
卢彩红
吕保先
龙剑文
Hu Huiqing;Li Jing;Fang Kun;Lu Caihong;LU Baoxian;Long Jianwen(Ninth Hospital of Wuhan,Wuhan 438000,China;The First Clinical Medical School of Hubei University of Chinese Medicine,Wuhan 430061,China)
出处
《中国中西医结合皮肤性病学杂志》
CAS
2019年第1期1-5,共5页
Chinese Journal of Dermatovenereology of Integrated Traditional and Western Medicine
