伸筋草提取物改善小鼠银屑病样皮损的作用机制

Effect And Mechanism of Lycopodium Clavatum Extract in Ameliorating Psoriatic Lesions of Mice

摘要

旨在探讨伸筋草提取物(LCE)对咪喹莫特(IMQ)诱导小鼠银屑病样皮损的改善作用及分子机制.采用IMQ建立小鼠银屑病模型,设置对照组、模型组、甲氨蝶呤组及LCE低、中、高剂量组,以银屑病面积与严重度指数(PASI)评分评价皮损程度,检测脾脏指数;苏木素伊红染色(HE)观察皮肤组织病理变化;实时荧光定量PCR(RT-qPCR)检测皮损组织p65、IκBα、p38、IL-17、IL-23、IL-6 mRNA表达;采用蛋白免疫印迹法(western blot)检测皮损组织p-p65、p65、p-IκBα、IκBα、p-p38、p38蛋白表达.结果显示:与模型组比较,LCE可显著改善红斑、鳞屑及浸润,降低PASI评分和脾脏指数(P<0.05);减轻表皮增厚与炎症细胞浸润;显著下调p65、IκBα、p38及炎症因子mRNA水平(P<0.05、0.01、0.001),并降低p-p65/p65、p-IκBα/IκBα、p-p38/p38蛋白比值(P<0.05、0.01),且呈剂量依赖性.结论:伸筋草提取物可有效改善IMQ诱导的小鼠银屑病样皮损,其机制可能与抑制NF-κB和p38 MAPK信号通路、降低炎症因子表达有关.

Abstract

The study aimed to investigate the therapeutic effects and underlying mechanisms of Lycopodium clavatum extract (LCE) on imiquimod (IMQ)-induced psoriasiform mouse models. IMQ-induced psoriasiform mice models were constructed. Psoriasis Area and Severity Index (PASI score) was used to evaluate the erythema, scaling, and infiltration of dorsal lesions in mice of each group after LCE intervention. The changes of spleen index of mice were recorded. Haematoxylin-eosin (HE) staining was used to examine the histological changes of skin lesions. Quantitative real-time PCR (RT-qPCR) was used to determine the levels of p65, IκBα, p38, IL-17, IL-23 and IL-6 mRNA in skin lesion. Western Blot (WB) was used to determine the levels of phosphorylated p65 (p-p65), p65, phosphorylated IκBα (p-IκBα), IκBα, phosphorylated p38 (p-p38) and p38 proteins in skin lesions. The experimental results demonstrated that LCE intervention could effectively improve erythema, scaling, and inflammatory infiltration, reduce PASI scores, and decrease the spleen indexes of psoriasiform mice (P<0.05). LCE significantly reduced the mRNA expression levels of p65, IκBα, p38, IL-17, IL-23, and IL-6 in skin lesion (P<0.05, 0.01, 0.001), as well as the protein expression levels of pp65/p65, p-IκBα/IκBα and p-p38/p38 in skin lesion (P<0.05, 0.01). The research indicated that LCE could effectively alleviates psoriasiform skin lesions of mice in a dose-dependent manner, and its protective effect might be related to the inhibition of nuclear factor-κB (NF-κB) and mitogen activated protein kinase (MAPK) signalling pathways.

关键词

伸筋草提取物 / 银屑病 / 咪喹莫特 / NF-κB / p38 MAPK / 炎症因子 / 信号通路

Key words

Lycopodium clavatum extract / psoriasis / immune factor / nuclear factor-κB / mitogen-activated protein kinase / signalling pathway

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基金资助

国家自然科学基金青年科学基金项目(81703409)