基于Spred-1蛋白信号通路探讨清血毒颗粒合剂对银屑病样小鼠的作用机制

Study on the Mechanism of Qingxuedu Granule Mixture on Psoriasis-like Mice Based on Spred-1 Protein Signaling Pathway

目的:观察清血毒颗粒合剂对咪喹莫特(IMQ)诱导的银屑病样小鼠模型的干预作用,并探讨其可能的治疗机制。方法:将32只雄性BALB/c小鼠随机分为正常组、模型组、清血毒颗粒组、复方青黛胶囊组;采用5%IMQ诱导的银屑病样小鼠模型。连续造模、给药7 d,第8天处死取材;实验过程观察小鼠银屑病皮损面积和严重程度指数(PASI)评分变化;并对相应样本进行HE染色观察皮损区组织病理变化;Western blot及RT-PCR法检测组织样本中Spred-1/Ras/ERK通路相关蛋白及mRNA表达。结果:与模型组相比,清血毒颗粒组、复方青黛胶囊组2组PASI评分、皮肤平均表皮厚度及病理Baker评分均下降明显(P<0.05),且与复方青黛胶囊组相比清血毒颗粒组下降较更明显(P<0.05)。IMQ造模后,模型组小鼠皮损区、近皮损区、非皮损区中Spred-1 mRNA表达降低(P<0.05);复方青黛胶囊治疗后三区域中Spred-1 mRNA变化无统计学意义(P>0.05),皮损区p-Ras、p-Raf1、p-ERK1/2、VEGF蛋白及Ras、Raf1、ERK1/2、VEGF的m RNA表达降低(P<0.001);而清血毒颗粒组小鼠三区域组织Spred-1 mRNA表达升高(P<0.05),皮损区p-Ras、p-Raf1、p-ERK1/2、VEGF蛋白及Ras、Raf1、ERK1/2、VEGF的m RNA表达降低(P<0.001)。结论:清血毒颗粒合剂可能通过促进Spred-1蛋白的表达,抑制Ras/ERK通路关键蛋白的磷酸化表达,继而改善IMQ诱导的银屑病样小鼠模型症状。

Objective:To observe the intervention effect and therapeutic mechanism of Qingxuedu Granules mixture on the psoriasis-like mouse model induced by imiquimod.Methods:Thirty-two male BALB/c mice were randomly divided into normal group,model group,Qingxuedu granule group,and compound Qingdai capsule group.A psoriasis-like mouse model induced by 5%IMQ.Continuous modeling and drug treatment for 7 days.Take a sample on the 8 th day.During the experiment,the PASI score changes and the pathological changes of the skin lesions were observed.Western blot and RT-PCR methods were used to detect Spred-1/Ras/ERK pathway related proteins and mRNA expression in tissue samples.Results:The PASI score,average skin thickness and pathological Baker score of the two groups in the Qingxuedu granule group and the compound Qingdai capsule group were lower than those in the model group(P<0.05).The Qingxuedu Granules group had a more significant decrease than the compound Qingdai capsule group(P<0.05).After IMQ modeling,the expression of Spred-1 mRNA in the skin lesion area,near-lesion area,and non-lesion area of the model group mice was reduced(P<0.05).There was no statistically significant change in Spred-1 mRNA in the three areas after treatment with compound Qingdai capsule(P>0.05).However,the expression of p-Ras,p-Raf1,p-ERK1/2,VEGF protein and the mRNA expression of Ras,Raf1,ERK1/2,and VEGF in the lesion area decreased(P<0.001).In the Qingxuedu Granules group,the expression of Spred-1 mRNA in the three regions of the mice was increased(P<0.05),and the lesions were p-Ras,p-Raf1,p-ERK1/2,VEGF protein and Ras,Raf1,ERK1/2.The mRNA expression of VEGF decreased(P<0.001).Conclusion:Qingxuedu Granules mixture can improve the symptoms of IMQ-induced psoriasis-like mouse model by promoting the expression of Spred-1 protein and inhibiting the phosphorylation expression of key proteins in the Ras/ERK pathway.