中药苦参水提物对慢性湿疹模型豚鼠炎症反应的影响

Effects of Sophora flavescens Extract on Inflammatory Reaction on Guinea Pig Models with Chronic Eczema

目的探讨中药苦参水提物对豚鼠慢性湿疹模型治疗效果。方法豚鼠24只,采用二硝基氯苯(2,4-dinitrochlorobenzene,DNFB)建立皮肤湿疹模型,随机分为3组,分别用生理盐水(模型对照组)、复方醋酸地塞米松软膏(阳性对照组)和苦参水提取物(药物治疗组),对湿疹部位皮肤连续涂抹给药14 d,以左、右耳质量之差来评价皮肤炎症肿胀度;流式细胞术检测外周血辅助性T(T helper,Th)细胞Th17(CD4+IL-17A^+T)的比例;qPCR检测耳片组织中维甲酸相关的孤儿受体γ(retinoic acid-related orphan receptorγ,RORγt)和IL-17AmRNA的相对表达量;Western Blot检测RORγt和IL-17A蛋白的表达;ELISA检测血清中促炎细胞因子白细胞介素-6(interleukin 6,IL-6)、肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)及抑炎细胞因子IL-10。结果药物治疗组与模型对照组相比豚鼠皮肤肿胀度明显减轻,差异具有统计学意义(P <0.05);与模型对照组相比,药物治疗组不仅降低外周血炎症细胞Th17所占比例,并且能显著降低转录因子RORγt)和IL-17AmRNA相对表达量(P <0.05,P <0.01);同时,RORγt和IL-17A蛋白也均明显下降(P <0.05)。血清中细胞促炎细胞因子IL-6和TNF-α明显降低(P <0.05),而抑炎细胞因子IL-10则显著上升(P <0.05)。结论苦参水提物能够调节Th17细胞的分化从而抑制炎症反应,有效治疗湿疹。

Objective To investigate the effects of extract from Chinese medicine Sophora flavescens on inflammatory reaction in Guinea pig models of chronic eczema. Methods The chronic eczema Guinea pig models were established by 2,4-dinitrochlorobenzene (DNFB) induction,and randomly divided into model control group,positive group and treatment group. Eczema skin was continuously administered with drugs for 14 days. The weight difference between the left and right ears was used to evaluate swelling, which reflects the degree of inflammation in the models. The percentage of Th17 cells in PBMC were analyzed by flow cytometry. The qPCR was used to measure the expression of RORγt and IL-17A mRNAs in ear tissue. Western Blot method was used to detect the expression levels of transcription factor RORγt and IL- 17A in tissue. The pro- inflammatory cytokines IL- 6, TNF- α and antiinflammatory cytokine IL-10 were detected by ELISA. Results Compared with model group,the treatment group of Sophora flavescens extract could significantly reduce the swelling on the eczema (P < 0.05). Similarly,the Sophora flavescens treatment not only significantly decreased the percentage of Th17 cells in PBMC (P < 0.05),but also reduced the mRNA levels of transcription factor RORγt and IL- 17A. Both RORγt and IL- 17A proteins were significantly reduced compared with model control. Furthermore, the levels of IL- 6 and TNF- α in serum of treatment groups were obviously down-regulated (P < 0.05,P < 0.01). However,the concentration of IL-10 in treatment group was obviously increased (P < 0.05). Conclusion Sophora flavescens extract could regulate the differentiation of Th17 cells and suppress the inflammatory reaction,intervene in the development of chronic eczema.