顽癣敌软膏调控JAK2/STAT3炎症信号通路治疗咪喹莫特诱导的小鼠银屑病作用机制研究

Study on the Mechanism of Wanxuandi Ointment in Treating Imiquimod-induced Psoriasis in Mice by Modulating JAK2/STAT3 Inflammatory Signaling Pathway

目的考察顽癣敌软膏(WXD)对银屑病的作用及机制。方法采用小鼠背部皮肤连续涂抹7 d咪喹莫特建立银屑病模型。时-效关系研究,设正常对照组,模型对照组,WXD第1、2、3、4天给药组及糠酸莫米松组,每日涂抹药物一次。量-效关系研究,设正常对照组,模型对照组,WXD 1、2、3次组及糠酸莫米松组,从造模第4天开始给药。通过银屑病皮损面积和严重程度指数(PASI)和病理观察进行药效评价;采用qRT-PCR法检测皮肤TNF-α和IL-1βmRNA的表达;ELISA法检测IL-23、IL-17A、5-HT、组胺含量;免疫组织化学法检测皮肤髓过氧化物酶(MPO)水平和p-JAK2表达;Western Blot法检测p-STAT3蛋白表达。结果时-效关系研究结果显示,WXD第1、2、3、4天给药组的PASI、棘层厚度、棘层杵状突起长度、棘层杵状突起面积/棘层面积均较模型组明显降低(P<0.05,P<0.01)。量-效关系研究结果显示,WXD 1次组效果更佳(P<0.05,P<0.01)。WXD可明显降低银屑病小鼠皮肤组织MPO、p-JAK2、p-STAT3蛋白表达及IL-23、IL-17A水平,降低皮肤TNF-α及IL-1βmRNA表达,降低血清5-HT和组胺水平(P<0.05,P<0.01)。结论顽癣敌软膏在造模同时进行预防给药和在银屑病症状明显时进行治疗给药的两种给药方式均有效,以每日给药1次治疗银屑病效果最佳,其机制与下调JAK2/STAT3信号通路抑制炎症反应有关。

Objective To investigate the effect and mechanism of Wanxuandi Ointment(WXD)on imiquimodinduced psoriasis in mice.Methods A psoriasis mice model was established by applying imiquimod to the hairless area on the back of the mice for 7 consecutive days.In the study of the time-effect relationship,mice were randomly separated into normal group,model group,groups of 1st,2nd,3rd,4th day of WXD-medication,and mometasone furoate(MF)group.The corresponding ointment was rubbed into the skin once daily.In the study of dose-effect relationship,mice were randomly separated into normal group,model group,groups of 1,2,3 time of WXD-medication,and MF group.These drugs were rubbed into the skin at the 4th day of modeling.The psoriasis area and severity index(PASI)and pathological observation were used to evaluate the effect of WXD on psoriasis.The mRNA expressions of TNF-αand IL-1βin the skin were detected by qRT-PCR.The content of IL-23,IL-17A,5-HT and histamine were detected by ELISA.The protein expressions of myeloperoxidase(MPO)and p-JAK2 in the skin were detected by immunohistochemistry.The protein expression of p-STAT3 was detected by Western Blot.Results The results of time-effect relationship showed that PASI,thickness of stratum spinosum,length of drumstick-shaped protrusion of stratum spinosum,area of drumstick-shaped protrusion of stratum spinosum/stratum spinosum area were obviously reduced in groups of 1st,2nd,3rd,4th day of WXD-medication(P<0.05,P<0.01).The results of dose-effect relationship suggested that all above mentioned indexes in group of 1 time of WXD-medication were also significantly decreased(P<0.05,P<0.01).Additionally,WXD significantly reduced the protein expression of MPO,p-JAK2,p-STAT3,IL-23 and IL-17A,and the mRNA expressions of TNF-αand IL-1βin skin tissues of psoriasis mice,as well as the serum levels of 5-HT and histamine(P<0.05,P<0.01).Conclusion WXD administration for prevention during the model establishment and therapeutic treatment of WXD in the stage of obvious psoriasis symptoms are effective,once-daily administration of WXD is considered be the best treatment for psoriasis.The possible mechanism is related to inhibiting inflammatory response by down-regulating JAK2/STAT3 signaling pathway.