蛇黄乳膏防治湿疹作用机理研究

Studies the Mechanism of Shehuang Cream on Eczema

目的:在确定蛇黄乳膏药理作用的基础上,进一步探索其作用机理,为其临床应用提供理论依据及实验依据。方法:选用40只雄性小鼠,对照组、模型组、蛇黄乳膏组和艾洛松组各10只,2 cm×2 cm范围腹部脱毛,正常组不致敏和激发,其余各组腹部脱毛区域行1%2,4-二硝基氟苯(DNFB)涂抹致敏,次日行1次加强致敏。以后每隔6 d行BDNF均匀涂抹各只小鼠右耳部激发,共激发3次。期间在其脱毛处,正常组和模型组乳膏基质涂抹,其余各组于脱毛处行对应乳膏1 mg·g-1均匀涂抹。采用HE染色后在显微镜下观察皮肤组织病理变化。采用ELISA检测血清IFN-γ、IL-4含量。免疫组化及Real-Time qPCR法分别检测小鼠皮损区域皮肤组织FLG、Caspase-14蛋白及基因表达。结果:与正常组相比,模型组表皮肥厚,角化过度,真皮层伴炎细胞浸润,存在慢性湿疹病理表现,蛇黄乳膏组和艾洛松组湿疹样病理表现得到改善。与正常组相比,模型组小鼠血清IFN-γ含量较高(P<0.05),IL-4含量较低(P<0.05);与模型组相比,蛇黄乳膏组和艾洛松组IFN-γ含量均降低(P<0.05),IL-4含量均升高(P<0.05)。与正常组相比,模型组小鼠皮损组织FLG、Caspase-14蛋白和mRNA表达量均较低(P<0.05);与模型组相比,蛇黄乳膏组和艾洛松组表现相似,皮损组织FLG、Caspase-14蛋白和mRNA表达量均上调(P<0.05)。结论:蛇黄乳膏可能通过调节IFN-γ、IL-4从而改善皮肤屏障FLG、Caspase-14表达发挥抗湿疹功效。

Objective:On the basis of determining the pharmacological action of ShehuangCream,the mechanism of action is further explored to provide a solid theoretical and experimental basis for its clinical application.Methods:40 male Swiss mice were randomly divided into control,model,Shehuang Cream and eloson groups,belly hair were removed 2×2 cm2 and 1%2,4-dinitrofluorobenzene(DNFB)smeared for sensitization in mice(except the controls),and the next day reduplicated it to strengthen the sensitization at the next day.6,12 and 18 days later the BDNF was applied evenly to the right ear of each mouse.Meanwhile mice were respectively smeared evenly the cream vehicle,Shehuang Cream or eloson on the hair removal area in mice.The histopathology of skin were checked under a microscope and the serum level of IFN-γand IL-4 were detected by Elisa.Immunohistochemistry and Real-Time qPCR method was used to detect the expression of FLG,Caspase-14 protein and gene in skin lesions of mice.Results:Compared with the normal group,the model group showed hyperkeratosis,dermis with inflammatory cell infiltration,and chronic eczema pathological manifestations.Eczema-like pathology was improved in the Shehuang Cream group and the eloson group.Compared with the normal group,the mice in the model group had higher serum levels of IFN-γ(P<0.05)and lower levels of IL-4(P<0.05).Compared with the model group,the content of IFN-γdecreased in both the Shehuang Cream group and eloson group mice(P<0.05),and the content of IL-4 increased(P<0.05).Compared with the normal group,the protein and mRNA expression of FLG,Caspase-14 in the model group were lower than that in the normal group(P<0.05).Compared with the model group,the protein and mRNA expression of FLG,Caspase-14 in the lesions were all increased in Shehuang Cream group and eloson group mice(P<0.05).Conclusion:ShehuangCream may play an anti-eczema effect by regulating IFN-γand IL-4 to improve the expression of skin barrier FLG and Caspase-14.