燥湿和胃方对脾胃湿热证大鼠HPA轴、炎症因子及免疫功能的影响

Effect of Zaoshi Hewei Formula on HPA Axis, Inflammatory Factors and Immune Function in Rats with Dampness Heat Syndrome of Spleen and Stomach

目的探讨燥湿和胃方对脾胃湿热证大鼠HPA 轴、炎症因子及免疫功能的影响。方法采用“环境因素+肥甘饮食+致炎因子”的方法复制脾胃湿热证动物模型。SPF 级SD 大鼠,♂,随机分为6 组,即正常组,模型组,三九胃泰组,燥湿和胃方高、中、低剂量组(4,2,1 g·kg^-1),每组10 只。采用放射免疫法检测促肾上腺皮质激素释放激素(corticotropinreleasing homone,CRH)、促肾上腺皮质激素(adrenocorticotropic hormone,ACTH)、皮质醇(cortisol,Cor)的含量;采用免疫组化法检测大鼠胃黏膜组织中环氧合酶-2(cyclooxygenase,COX-2)表达水平;采用ELISA 法检测肿瘤坏死因子-α(TNF-α)、白介素1β(IL-1β)、干扰素-γ(interferen-gamma,IFN-γ)、白介素4(IL-4)含量。结果与正常组相比,模型组大鼠血清中CRH、ACTH、Cor、TNF-α、IL-1β、IFN-γ、IL-4、IFN-γ/IL-4 值明显升高(P<0.05)。与模型组比较,燥湿和胃方各剂量组均显著降低大鼠血清中上述指标的含量(P<0.05)。与燥湿和胃方高剂量组相比,低剂量组大鼠血清中CRH、ACTH、Cor含量明显较高(P<0.05);中剂量组大鼠血清CRH、ACTH、Cor 含量稍高,但差异没有统计学意义,IL-1β、IL-4 与其相当,差异没有统计学意义,TNF-α、COX-2的改善程度明显较弱(P<0.05),IFN-γ、IFN-γ/IL-4 值均明显偏高(P<0.05)。结论燥湿和胃方能有效改善脾胃湿热模型大鼠的疾病活动状态,该作用的发挥可能与调整HPA 轴CRH、ACTH、Cor 的含量有关,可能通过降低HPA 轴相关指标,降低机体炎症反应,改善免疫状态,恢复免疫系统Th1/Th2的动态平衡。

OBJECTIVE To study the effects of Zaoshi Hewei Formula on HPA axis,inflammatory factors and immune function in rats with dampness heat syndrome of spleen and stomach.METHODS The animal model of damp heat syndrome of spleen and stomach was replicated by "environmental factors + fat diet + inflammatory factors".SPF SD male rats were randomly divided into 5 groups: normal group,model group,Zaoshi Hewei Formula high,middle and low dose groups(4,2, 1 g·kg^-1).The content of adrenocorticotropic hormone releasing hormone (corticotropin releasing homone,CRH),corticotropin (adrenocorticotropic hormone,ACTH) and cortisol(Cor) were detected by radioimmunoassay.Immunohistochemistry was used to detect cyclic oxygenase-2 (cyclooxygenase,COX-2) in rat gastric mucous membrane.The expression level of tumor necrosis factor-α(TNF-α),rat interleukin 1β(IL-1β),interferon-γ(interferen-gamma,IFN-γ) and rat interleukin 4 (IL-4) were detected by ELISA.RESULTS Compared with the normal group,the serum levels of CRH,ACTH,Cor,TNF-α,IL-1β,IFN-γ,IL-4 and IFN-γ/IL-4 in the rat serum in the model group were significantly increased(P<0.05).Compared with the model group,the above indexes in the serum of rats were significantly reduced by Zaoshi Hewei formula(P<0.05).Compared with the high dose of Zaoshi Hewei Formula group,the content of CRH,ACTH and Cor in the low dose group was significantly higher(P<0.05).The content of each index in the middle dose group was slightly higher than that of the high dose group,and there was no significant difference between the groups.The improvement of TNF-α and COX-2 was significantly lower than that of high dose group (P<0.05),but there was no significant difference of the IL-4 index between the middle dose group and high dose group,and the values of IFN-γ/IL-4 in the middle dose group were significantly higher than that of high dose group (P<0.05).CONCLUSION Zaoshi Hewei Formula can effectively improve the state of disease activity in rat model of dampness heat syndrome of spleen and stomach,which may be related to adjusting the content of CRH,ACTH and Cor of HPA axis by reducing the related indexes of HPA axis,reducing the inflammatory response of the body,improving the immune state and restoring the dynamic balance of Th1/ Th2 in the immune system.