石军颗粒治疗肝性脑病的作用机制研究

Study on the mechanism of Shijun granules in the treatment of hepatic encephalopathy

目的 探讨石军颗粒治疗肝性脑病(HE)的作用机制。方法 首先,通过数据库筛选并预测石军颗粒化学成分相关靶点和HE相关靶点;然后,构建石军颗粒成分靶点网络图以及石军颗粒和肝性脑病靶点蛋白-蛋白相互作用(PPI)网络图,并进行PPI网络融合以获得石军颗粒治疗HE的作用靶点;随后,进行KEGG通路富集分析;最后,建立PC12细胞损伤模型和HE相关大鼠模型,实验分为正常组、模型组、对照组和给药组,在细胞和动物水平对石军颗粒的作用机制进行验证。结果 筛选得到石军颗粒20个化学成分和99个作用靶点,获得305个HE相关靶点。之后进行网络图构建和PPI网络融合,获得207个石军颗粒治疗HE的候选靶点。KEGG通路富集分析结果显示,石军颗粒可能是通过MAPK、PI3K-Akt和Neurotrophin等信号通路发挥治疗HE的作用。最后,通过体内和体外实验验证明确石军颗粒通过调控MAPK和PI3K-Akt信号通路发挥作用。同时,石军颗粒降低了HE大鼠血清中谷丙转氨酶、谷草转氨酶、碱性磷酸酶、谷氨酰转移酶、球蛋白的含量和动脉血氨浓度。结论 石军颗粒通过调控MAPK和PI3K-Akt信号通路改善肝性脑病患者的肝功能,降低由氨诱导的脑细胞肿胀和氧化应激,发挥治疗肝性脑病的作用。同时,动物实验表明,石军颗粒可有效改善HE大鼠肝功能,降低动脉血氨浓度。

Objective To explore the molecular mechanisms of Shijun granules in the treatment of hepatic encephalopathy(HE). Methods First,the chemical constituents and targets in Shijun granules and HE related targets were screened and predicted. Then,the component target network diagram of Shijun granules and the protein-protein interaction(PPI)network diagram of Shijun granules and hepatic encephalopathy target were constructed,and the PPI network was fused to obtain the target of Shijun granules in the treatment of HE. Then KEGG enrichment pathway analysis was carried out. Finally,a PC12 cell injury model and a rat model related to HE were established. The experiments were divided into the normal group,the model group,the control group and the administration group,to verify the mechanism of action of Shijun granules at the cellular and animal levels. Results A total of 20 chemical constituents and 99 action targets of Shijun granules,and 305 hepatic encephalopathy disease targets were obtained. Network graph construction and PPI network fusion were carried out to obtain 207 candidate targets of Shijun granules for the treatment of HE. The results of KEGG pathway enrichment analysis indicated that Shijun granules may play a role in the treatment of HE by regulating some pathways including MAPK signaling pathway,PI3K-Akt signaling pathway,and neurotrophin signaling pathway. The in vivo andf in vitro experimental results indicated that Shijun granules play a role in regulating MAPK and PI3K-Akt signaling pathways. In addition,Shijun granules reduced ALT,AST,ALP,GGT,G content and blood ammonia concentration in HE rats. Conclusion Shijun granules can improve liver function in patients with HE and reduce brain cell swelling and oxidative stress induced by ammonia by regulating the MAPK and PI3K-Akt signaling pathways,and play a role in the treatment of HE. Meanwhile,animal experiments show that Shijun granules can effectively improve the liver function of rats with HE and reduce the blood ammonia concentration.