调控JAKS/STATS通路探讨健脾祛湿法治疗寻常型银屑病的机制

Mechanism of Jianpi Qushi Decoction in Treating Psoriasis by Regulating JAKS/STATS Pathway

目的通过健脾解毒汤及拆方干预脾虚湿盛寻常型银屑病小鼠,观察转录因子(STAT)3、磷酸化转录因子(p-STAT)3、STAT5、p-STAT5的表达水平,发现健脾祛湿法治疗银屑病的作用机制。方法健康雄型美国癌症研究所(ICR)小鼠70只,4~6周龄;体质量18~25 g,平均20.4 g。将小鼠分为7组,分别为空白对照组(A组)、阿维A胶囊组(B组)、健脾汤组(C组)、解毒汤组(D组)、健脾解毒汤组(E组)、脾虚模型组(F组)、脾虚+银屑病组(G组),每组10只。除空白对照组外,其余各组进行大黄液灌胃,建立脾虚模型。然后除空白对照组和脾虚模型组外,其余各组再进行咪喹莫特软膏诱导银屑病。在造模45 d后,处死小鼠,对小鼠皮损组织进行收集,通过实验荧光定量PCR(RTqPCR)验证STAT3和STAT5的表达水平,蛋白免疫印迹(Western blot)分别验证STAT3,p-STAT3,STAT5,p-STAT5蛋白表达水平,比较不同药物干预后,各指标变化。计量资料以均数±标准差(Mean±SD)表示,组间比较采用单因素方差分析,两两比较采用最小显著性差异法(LSD)。结果E组STAT3和STAT5在mRNA水平的表达量明显低于C组和D组(P<0.05),在蛋白水平上,STAT3和STAT5在各组之间差异无统计学意义(P>0.05)。E组p-STAT3和p-STAT5的蛋白表达水平明显低于C组和D组,差异有统计学意义(P<0.05)。结论健脾解毒汤可以明显调节STAT3,STAT5,p-STAT3,p-STAT5的表达水平,可能是健脾祛湿法治疗银屑病的机制之一。

Objective In order to observe the expression levels of signal transducer and activator of transcription 3(STAT3);phosphorylated signal transducer and activator of transcription 3(p-STAT3);signal transducer and activator of transcription 5(STAT5);phosphorylated signal transducer and activator of transcription 5(p-STAT5)in mice with spleen deficiency and dampness-filled psoriasis vulgaris by Jianpi Jiedu decoction and disinfecting decoction,and to discover the mechanism of Jianpi Qushi therapy for psoriasis.Methods Seventy healthy male(ICR)mice were aged from four to six weeks.The average weight was 20.4 g,with a range of 18 to 25 g.Mice were divided into seven groups:blank control group(group A),spleen deficiency model group(group B),spleen deficiency+psoriasis group(group C),Awei A capsule group(group D),Jiedu decoction group(group E),Jianpi decoction group(group F)and Jianpi Jiedu decoction group(group G),with 10 mice in each group.In addition to the blank control group,the other groups were given rhubarb liquid gavage to establish spleen deficiency models.Then imiquimod ointment was used to induce psoriasis except blank control group and spleen deficiency model group.After forty days of modeling,mice were sacrificed and skin tissue was collected.The expression levels of STAT3 and STAT5 were verified by RT-qPCR,and the protein expression levels of STAT3,p-STAT3,STAT5 and p-STAT5 were verified by Western blotting,respectively,and the changes of various indicators were compared after different drug interventions.Measurement data were expressed as mean±standard deviation(Mean±SD),one-way anova was used for comparison anmong groups,and least significant difference method(LSD)was used for pairwise comparison.Results The mRNA levels of STAT3 and STAT5 in Jianpi Jiedu decoction group were significantly lower than those in Jianpi Jiedu decoction group and Jiedu decoction group(P<0.05).The protein expression levels of p-STAT3 and p-STAT5 in Jianpi Jiedu decoction group were significantly lower than those in Jianpi Jiedu decoction group and Jiedu decoction group,with significant statistical significance(P<0.05).Conclusion Jianpi Jiedu decoction could significantly regulate the expression levels of STAT3,STAT5,p-STAT3 and p-STAT5,may be one of the mechanisms of Jianpi Qushi therapy for psoriasis.