基于miR-155-SOCS1-JAK2/STAT3与Treg细胞通路的相关性研究蜈蚣败毒饮治疗银屑病机制

Study of the Mechanism of Wugong Baidu Yin(蜈蚣败毒饮) in the Treatment of Psoriasis Based on the Correlation Between miR-155-SOCS1-JAK2/STAT3 and Treg Cell Pathway

目的研究中药制剂蜈蚣败毒饮对银屑病模型的皮损疗效,在血清学层面上分析其对于miR-155-SOCS1-JAK2/STAT3介导下Treg细胞主导通路IL-2-Foxp3-IL-10/TGF-β的干预作用,并分析二者的相关性。方法将30只大鼠随机分为生理盐水空白组,生理盐水模型组,甲氨蝶呤西药对照组,蜈蚣败毒饮低、中、高剂量组,除空白组外采用咪喹莫特法进行建模,再以相应的药液进行灌胃。采用鼠银屑病皮损面积和严重程度指数(mice psoriasis area and severity index,MPASI)评分法评价灌胃前后银屑病模型鼠的皮损指数,并计算出其变化量(△MPASI),采用聚合酶链式反应(polymerase chain reaction,PCR)法检测血清中微小核糖核酸-155(miR-155)mRNA、细胞因子信号抑制物1(suppressor of cytokine signaling 1,SOCS1)mRNA、JAK2 mRNA、STAT3 mRNA的表达量,采用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测血清中白细胞介素-2(interleukin-2,IL-2)、叉头框蛋白3(forkhead box protein 3,Foxp3)、血清中白细胞介素-10(interleukin-10,IL-10)、转化生长因子-β(transforming growth factor-β,TGF-β)的含量,同时,整体评估△MPASI与各因子表达之间的相关性,从而系统评价皮损疗效与两大通路的关系。结果甲氨蝶呤及蜈蚣败毒饮各剂量组均能够降低银屑病鼠模型的皮损指数(P<0.05或P<0.01),以甲氨蝶呤和蜈蚣败毒饮高剂量的疗效最为显著(P<0.01);甲氨蝶呤和蜈蚣败毒饮高剂量组可以显著下调miR-155 mRNA(P<0.01),上调SOCS1 mRNA(P<0.01),抑制JAK2/STAT3的经典通路(P<0.01);除IL-2外(P>0.05),蜈蚣败毒饮高剂量组和甲氨蝶呤可以显著上调血清中Foxp3、IL-10、TGF-β的含量(P<0.01);血清中△MPASI与miR-155、两面神激酶2(janus kinase 2,JAK2)、信号转导及转录激活蛋白3(signal transducer and activator of transcription 3,STAT3)呈负相关(P<0.05),△MPASI与SOCS1、Foxp3、IL-10、TGF-β呈正相关(P<0.05)。结论蜈蚣败毒饮的作用途径可能不依赖于IL-2的刺激,其可能通过干预miR-155-SOCS1-JAK2/STAT3通路,通过旁路刺激影响Treg细胞的转录和分泌因子而发挥银屑病治疗的作用。

Objective To study the effect of Wugong Baidu Yin(蜈蚣败毒饮)on psoriasis model,to analyze the intervention effect of Wugong Baidu Yin on Treg cell dominant pathway IL-2-Foxp3-IL-10/TGF-βmediated by miR-155-SOCS1-JAK2/STAT3 at the serological level,and analyze the correlation between them.Methods 30 mice were randomly divided into blank group,model group,methotrexate group,low,medium and high dose groups of Wugong Baidu Yin,except the blank group,Imiquimod method was used for modeling,then the corresponding solution was given by gavage.MPASI method was used to evaluate the skin lesion index of psoriasis model mice before and after intragastric administration,and calculate the change(△MPASI),the expressions of miR-155 mRNA,SOCS1 mRNA,JAK2 mRNA and STAT3 mRNA in serum were detected by PCR method,the levels of IL-2,Foxp3,IL-10 and TGF-βin serum were detected by ELISA method,at the same time,the correlation between△MPASI and the expression of each factor was evaluated,so as to systematically evaluate the relationship between the curative effect of skin lesions and the two pathways.Results Methotrexate group and each dose group of Wugong Baidu Yin could reduce the skin lesion index of psoriasis model mice(P<0.05 or P<0.01),the high dose of Wugong Baidu Yin and methotrexate were the most effective(P<0.01);the high dose of Wugong Baidu Yin and methotrexate group could significantly down regulate miR-155 mRNA(P<0.01),up regulate SOCS1 mRNA(P<0.01),and inhibit JAK2/STAT3 classical pathway(P<0.01);except IL-2(P>0.05),the high dose of Wugong Baidu Yin and methotrexate could significantly up regulate the contents of Foxp3,IL-10 and TGF-βserum(P<0.01);there was a negative correlation between△MPASI and miR-155,JAK2,STAT3 in serum(P<0.05),meanwhile,there was a positive correlation between△MPASI and SOCS1,Foxp3,IL-10,TGF-βin serum(P<0.05).Conclusion The mechanism of action of Wugong Baidu Yin may not depend on the stimulation of IL-2,it may interfere with miR-155-SOCS1-JAK2/STAT3 pathway and affect the transcription and secretion of Treg cells through the pathway stimulation to play the role of psoriasis treatment.