郁金散对大肠湿热证大鼠HPA轴的影响机制

基于分子对接并辅以生物学验证探讨郁金散治疗大肠湿热证HPA轴损伤的机制。采用高效液相色谱法(High Performance Liquid Chromatography,HPLC)检测郁金散的主要活性成份,用分子对接观察其活性成分与HPA轴相关激素、蛋白之间的结合力,通过网络药理学分析构建药物靶标网络,预测郁金散活性成分作用的主要靶标,并建立大肠湿热证大鼠模型,进行生物学验证。计算各组大鼠肾上腺指数,观察肾上腺组织变化特征,检测关键靶标及其下游激素的表达量。分子对接与网络分析可以推测CRH可能是调控HPA轴亢进的潜在蛋白,盐酸小檗碱、没食子酸、大黄素可能是郁金散调节HPA轴亢进的活性成分。生物学验证实验结果表明,与空白对照组相比,模型组和自愈组大鼠肾上腺指数及血清中CRH、ACTH、CORT的含量显著或极显著升高(P<0.05或P<0.01),模型组大鼠肾上腺出现一定的病理变化。经郁金散治疗后,以上各指标及病理变化均有所改善,其中以郁金散高剂量组治疗效果最佳,验证了分子对接的部分预测结果。研究证实郁金散可能是通过下调CRH的水平调节大肠湿热证大鼠HPA轴功能亢进,为进一步开展郁金散治疗HPA轴亢进的机制研究提供了新思路和新方法。

Mechanism of Yujin powder on HPA axis in rats with Large Intestine Damp-heat syndrome

Abstract: Study on mechanism of Yu Jin power(YJP) in treating hyperfunction of HPA axis with large intestine damp-heat syndrome that based on molecular docking and biological verification.The main active components of YJP were determined by High Performance Liquid Chromatography. We observed the binding force between the active components and HPA axis related hormones and proteins by molecular docking. Drug target network was constructed by network pharmacology analysis to predict the main target of active components of YJP, and the rat model of large intestine damp-heat syndrome was established for biological verification. The adrenal gland index of rats in each group was calculated, the change characteristics of its tissue were observed, and the expression levels of key targets and their downstream hormones were detected. Molecular docking and network analysis suggested that CRH may be a potential protein to regulated HPA axis , and berberine , gallic acid, emodin may be the active components of YJP . The results of biological verification experiment showed that compared with normal control group, the adrenal gland index and the contents of serum CRH, ACTH and CORT in Model group and Self-healing group were significantly or extremely significantly increased (P< 0.05 or P< 0.01). Some pathological changes were observed in adrenal gland of model group. After treatment with YJP, all the above indexes and pathological changes were improved, and the high-dose Group of YJP had the best therapeutic effect. This study confirmed that YJP may regulate the HPA axis hyperfunction in rats with large intestinal damp-heat syndrome by reducing the level of CRH, which provides new ideas and new methods for further research on the mechanism of YJP in the treatment of HPA axis hyperactivity.