去氢茯苓酸对湿疹大鼠的治疗作用及对免疫反应的影响

Therapeutic effect of dehydropachymic acid on rats with eczema and its effect on immune response

为探讨去氢茯苓酸(dehydropachymic acid,DPA)对湿疹大鼠的治疗作用及对免疫反应的影响,构建二硝基氯苯(dinitrochlorobenzene,DNCB)诱导的湿疹大鼠模型,应用不同浓度的DPA(10、40和80 mg/kg)治疗大鼠14 d,期间复方甘草酸苷(compound glycyrrhizin,CG)(15 mg/kg)作为阳性对照药物。H-E染色观察大鼠皮肤组织学形态;ELISA检测各组大鼠血清IL-4和IFN-γ的水平及血清IgA、IgM和IgG水平。共培养刀豆蛋白A(Concanavalin A,ConA)诱导的淋巴细胞与人角质形成细胞HaCaT,应用40μg/mL DPA处理细胞。MTT法检测HaCaT细胞的增殖能力,TUNEL法检测HaCaT细胞的凋亡情况。H-E染色结果显示,DPA以剂量依赖性方式减轻湿疹大鼠的皮损症状。DPA降低了湿疹大鼠的血清IL-4、IgA、IgM和IgG水平,并提高了IFN-γ的水平(均P<0.05)。中剂量(40 mg/kg)和高剂量(80 mg/kg)DPA对湿疹大鼠的改善作用与15 mg/kg剂量CG药效相当。DPA未影响HaCaT细胞的增殖(P>0.05);然而,DPA抑制了淋巴细胞共培养HaCaT细胞的凋亡(P<0.05)。由此,DPA通过调节Th1/Th2平衡影响机体免疫功能,减轻湿疹相关病变。

In order to evaluate the therapeutic effect of dehydropachymic acid(DPA) on eczema rats and its effect on immune response,a rat model of eczema induced by dinitrochlorobenzene(DNCB) was established.Different concentrations of DPA(10,40,and 80 mg/kg) were used to treat eczema rats for fourteen days,and compound glycyrrhizin(CG)(15 mg/kg) was used as the positive treatment control.H-E staining was used to observe the histopathological morphology of rat skin tissues.ELISA was used to detect the levels of serum IL-4 and IFN-γ as well as IgA,IgM and IgG.Concanavalin A(ConA)-activated lymphocytes were co-cultured with human keratinocyte HaCaT,followed by treatment of 40 μg/mL DPA.The proliferation of HaCaT cells was detected by MTT method,and the apoptosis of HaCaT cells was detected by TUNEL method.H-E staining outcomes showed that DPA relieved the skin lesions of eczema rats in a dose-dependent manner.DPA decreased the levels of serum IL-4,IgA,IgM and IgG in rats with eczema,and increased the level of IFN-γ(all with P<0.05).The reliefs of eczema in middle-dose(40 mg/kg) and high-dose(80 mg/kg) DPA-treated rats were equivalent to that of 15 mg/kg CG-treated positive control rats.DPA did not affect the proliferation of HaCaT cells(P>0.05).However,DPA inhibited the apoptosis of HaCaT cells co-cultured with lymphocytes(P<0.05).Therefore,DPA regulates the immune function of the eczema rats by improving the Th1/Th2 balance,thereby alleviating eczema-related lesions.