基于网络药理学和实验验证探讨香连化浊方对慢性萎缩性胃炎的作用机制

Mechanism of Xianglian Huazhuo Prescription Against Chronic Atrophic Gastritis Based on Network Pharmacology and Experimental Verification

目的基于网络药理学及动物实验探讨香连化浊方治疗慢性萎缩性胃炎(CAG)的作用机制,为临床推广应用提供科学依据。方法基于网络药理学预测得到香连化浊方治疗CAG的可能作用靶点和通路,采用水杨酸钠、N-甲基-N′-硝基-N-亚硝基胍(MNNG)及饥饱失常多因素诱导大鼠CAG模型,给予香连化浊方和摩罗丹干预60 d。给药结束后处死大鼠,苏木素-伊红(HE)法观察各组胃黏膜组织形态学变化;酶联免疫吸附测定法(ELSIA)检测大鼠血清中白细胞介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、白细胞介素-1β(IL-1β)、血管内皮生长因子(VEGF)的含量;免疫组织化学法(IHC)检测各组胃黏膜B细胞淋巴瘤-2(Bcl-2)家族抗凋亡因子(Bad)、细胞抗凋亡因子Bcl-2蛋白表达含量。结果最终获得香连化浊方潜在活性成分241种,核心靶点53个。香连化浊方可影响细胞增殖凋亡、炎症反应、DNA代谢过程的调控、细胞对氧化还原的反应等多个生物过程,影响磷脂酰肌醇3-激酶/蛋白激酶B(PI3K/Akt)信号通路、TNF信号通路、丝裂原活化蛋白激酶(MAPK)信号通路、癌症及癌症相关等多个信号通路。动物模型验证结果,香连化浊方能够降低CAG大鼠血清中IL-6、TNF-α、IL-1β、VEGF水平;降低胃组织中Bad、Bcl-2蛋白表达水平。结论香连化浊方可通过参与细胞增殖、凋亡和炎症反应等生物过程,调控PI3K/Akt信号通路,改善CAG胃黏膜损伤。

Objective To explore the mechanism of Xianglian Huazhuo prescription in the treatment of chronic atrophic gastritis(CAG)based on network pharmacology and animal experiments,so as to provide scientific basis for clinical application.Method The possible targets and pathways of Xianglian Huazhuo prescription in the treatment of CAG were obtained based on the prediction of network pharmacology.The CAG rat model was induced by sodium salicylate,N-methyl-N'-nitro-N-nitrosoguanidine(MNNG)and hunger and satiety disorder.Then the CAG rats were treated with Xianglian Huazhuo prescription and morodan for 60 days.After administration,the rats were sacrificed,and the content of interleukin-6(IL-6),tumor necrosis factor-α(TNF-α),interleukin-1β(IL-1β)and vascular endothelial growth factor(VEGF)in serum was determined by enzyme linked immunosorbent assay(ELISA).In addition,the protein expression of Bad and Bcl-2 in gastric mucosa was detected by immunohistochemistry(IHC).Result A total of 241 active components of Xianglian Huazhuo prescription and 53 core targets were obtained.Xianglian Huazhuo prescription affected multiple biological processes,such as cell proliferation and apoptosis,inflammatory reaction,regulation of DNA metabolism,and cell response to redox,as well as phosphatidylinositol 3-kinase/protein kinase B(PI3K/Akt),TNF,mitogen-activated protein kinase(MAPK),cancer and cancer-related signaling pathways.The animal model verification showed that Xianglian Huazhuo prescription lowered the levels of IL-6,TNF-α,IL-1βand VEGF in serum of CAG rats,and reduced the protein expression of Bad and Bcl-2 in gastric tissue.Conclusion Xianglian Huazhuo prescription could regulate PI3K/Akt signal pathway and improve gastric mucosal injury in CAG by participating in biological processes such as cell proliferation,apoptosis and inflammation.