基于IL-23/IL-17炎症轴探讨玉屏风颗粒治疗CU大鼠的效应机制

Mechanism of Yupingfeng Granule on CU Rats Based on IL-23/IL-17 Inflammation Axis

目的:研究玉屏风颗粒对慢性荨麻疹(CU)大鼠皮肤肥大细胞脱颗粒的影响及白细胞介素-23(IL-23),白细胞介素-17(IL-17)炎症轴的干预机制。方法:选用SPF级SD大鼠,取60只随机分为正常组,模型组,氯雷他定组(0.9 mg·kg^-1·d^-1),玉屏风颗粒高、中、剂量组(4.05,2.7,1.35 g·kg^-1·d^-1)。运用腹腔注射卵白蛋白与氢氧化铝悬液和百白破疫苗进行致敏复制CU大鼠模型。苏木素-伊红(HE)染色观察大鼠皮肤组织病理学改变;甲苯胺蓝染色观察大鼠皮肤组织肥大细胞脱颗粒现象;免疫组化(IHC)检测皮肤组织IL-23,IL-17蛋白表达;采用实时荧光定量聚合酶链式反应(Real-time PCR)法检测皮肤组织IL-23,IL-17 mRNA转录水平。结果:玉屏风颗粒可显著改善CU大鼠皮肤真皮水肿、胶原束距离增宽、炎性细胞浸润等病理表现,同时可减轻CU大鼠皮肤组织肥大细胞脱颗粒反应。与正常组比较,模型组CU大鼠皮肤的IL-23,IL-17 mRNA水平及蛋白表达评分均明显增加(P<0.05,P<0.01);与模型组比较,玉屏风颗粒能显著下调CU大鼠皮肤的IL-23 mRNA水平及蛋白表达评分(P<0.05,P<0.01),以高剂量组疗效最佳;玉屏风颗粒对IL-17无显著调控作用。结论:玉屏风颗粒可显著减轻CU大鼠皮肤组织肥大细胞脱颗粒,改善真皮水肿、浆液性渗出、炎性细胞浸润等病理表现,其机制可能与其抑制IL-23促炎细胞因子分泌而改善CU病变有关。

Objective:To study the effect of Yupingfeng granule on the degranulation of skin mast cells in chronic urticaria(CU)rats and the intervention mechanism of interleukin-23(IL-23),interleukin-17(IL-17)inflammation axis.Method:Totally 60 SPF SD rats were selected and randomly divided into normal group(normal saline),model group(normal saline),and loratadine group(0.9 mg·kg^-1·d^-1),high-dose Yupingfeng granules group(4.05 g·kg^-1·d^-1),middle-dose group(2.7 g·kg^-1·d^-1),low-dose group(1.35 g·kg^-1·d^-1).The CU rat model was reproduced through intraperitoneal injection of ovalbumin with aluminum hydroxide suspension and DTP vaccine.Histopathological changes of rat skin were observed by hematoxylin-eosin(HE)staining.Degranulation of mast cells in rat skin was determined by toluidine blue staining.IL-23 and IL-17 protein expressions in skin tissue were determined by immunohistochemistry(IHC).IL-23 and IL-17 mRNA transcription levels in skin tissue were determined by real-time fluorescence quantitative polymerase chain reaction(Real-time PCR).Result:Yupingfeng granules can significantly alleviate the pathological manifestations of dermal edema,collagen beam distance,inflammatory cell infiltration of CU rats,and reduce the degranulation reaction of skin tissue mast cells in CU rats.The IL-23,IL-17 mRNA and protein expressions of the skin of model group were significantly increased compared with the normal group(P<0.05,P<0.01).Compared with the model group,Yupingfeng granules can significantly down-regulate IL-23 mRNA and protein expressions of CU rats(P<0.05,P<0.01).Yupingfeng granules had no significant regulatory effect on IL-17.Conclusion:Yupingfeng granule can significantly reduce the degranulation of mast cells in skin tissue of CU rats,and improve the pathological manifestations,such as dermal edema,serous exudation and inflammatory cell infiltration.The mechanism may be related to inhibiting the secretion of IL-23 pro-inflammatory cytokines and improving CU lesions.