Abstract: Background Gastric cancer，as one of the common malignant tumors of the digestive tract，has a complex pathogenesis and limited therapeutic means，and the mechanism of Modified Qifang Weitong Granules for the treatment of gastric cancer is still not completely clear. Objective To investigate whether Modified Qifang Weitong Granules inhibit the proliferation of gastric cancer through glycolysis mediated by phosphatidylinositol 3-kinase（PI3K）/protein kinase B（AKT）/hypoxia-inducible factor 1 subunit ɑ（HIF-1ɑ）pathway. Methods From March 2024 to June 2025，50 nude mice were selected for HGC-27 gastric cancer cell transplantation tumor model preparation，and 40 were finally included in the study. Forty nude mice were randomly divided equally into control group，Modified Qifang Weitong Granules low ，medium and high dose group，and 10 rats in each group. They were respectively given intragastric administration of the drug solutions at doses of 15.28g·kg-1·d-1，30.55 g·kg-1·d-1，61.1 g·kg-1·d-1. The low，medium and high dose groups were given the corresponding dose of drug solution by gavage，and the control group was given an equal volume of saline by gavage once a day. After 14d of drug administration，the weight of transplanted tumor was weighed to calculate the tumor suppression rate，the transplanted tumor tissues were taken for Hematoxylin-Ihon（HE） staining and immunohistochemistry analysis，the Microscale assay kit detected the production of Adenosine triphosphate（ATP） and lactate in the transplanted tumor tissues，and real-time quantitative polymerase chain reaction（RT-qPCR） detected the expression of HK2 and LDHA mRNA in the transplanted tumor tissues. Network pharmacology analyzed the key targets and pathways of Modified Qifang Weitong Granules in regulating glycolysis in gastric cancer cells，and detected the changes in the expression of proteins related to the PI3K/AKT/HIF-1ɑ pathway by Western blot. Results After 14d of intervention，the transplanted tumor mass of the Modified Qifang Weitong Granules low，medium and high dose groups were lower than that of the control group（P<0.05），and the tumor suppression rates were 11.90%，33.61% and 52.70%，respectively； HE staining showed that different degrees of tumor cell necrosis and cytosolic fragmentation could be seen in the low，medium and high dose groups of Modified Qifang Weitong Granules compared with the control group；immunohistochemistry showed that the expression of GLUT1 and PFK1 proteins in the transplanted tumors of the low-，medium and high-dose groups of Jiawei Qifang Gastroparesis Granules was lower than that of the control group（P<0.05）；The results of the Microscale assay kit and RT-qPCR showed that the expression of ATP，lactate，HK2 and LDHA mRNA in the transplanted tumors of the low，medium and high dose groups of Modified Qifang Weitong Granuleswas lower than that of the control group （P<0.05）； the network pharmacological analysis showed that the PI3K/AKT/HIF-1ɑ signaling pathway was closely related to the regulation of glycolysis in gastric cancer cells by Modified Qifang Weitong Granules. Western Blot analysis showed that the p-PI3K/PI3K protein ratio and HIF-1ɑ protein expression in the transplanted tumor tissues of the Modified Qifang Weitong Granules low，medium and high dose groups were lower than those in the control group. Conclusion Modified Qifang Weitong Granule can inhibit the growth of transplanted tumors in HGC-27 homozygous nude mice，the mechanism of which is related to the inhibition of the activation of the PI3K/AKT/HIF-1ɑ pathway，the reduction of the expression of GLUT1，PFK1，HK2 and LDHA，the reduction of ATP and lactic acid production，and the regulation of glycolysis.

Key words: Gastric Cancer, Modified Qifang Weitong Granules, Glycolysis, PI3K/AKT/HIF-1ɑ, Animal Experimentation

摘要： 背景 胃癌作为常见的消化道恶性肿瘤之一，其发病机制复杂且治疗手段有限，加味七方胃痛颗粒治疗胃癌的机制仍不完全明确。目的 探讨加味七方胃痛颗粒通过磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（AKT）/缺氧诱导因子1α（HIF-1ɑ）介导肿瘤细胞糖酵解抑制胃癌增殖的作用机制。方法 2024年3月—2025年6月，选择50只裸鼠建立HGC-27胃癌细胞移植瘤模型，最后纳入研究40只。随机分为对照组、加味七方胃痛颗粒低、中、高剂量组，每组 10 只，分别予 15.28 g·kg-1·d-1、30.55 g·kg-1·d-1 和 61.1 g·kg-1·d-1 剂量药液灌胃，对照组给与等体积生理盐水灌胃，1 次 /d。给药 14 d 后，称量移植瘤重量计算抑瘤率，进行移植瘤组织苏木精 - 伊红（HE）染色及免疫组化分析，微量法检测移植瘤组织中三磷酸腺苷（ATP）、乳酸的生成量，实时荧光定量聚合酶链式反应（RTqPCR）检测移植瘤组织中己糖激酶 2（HK2），乳酸脱氢酶 A（LDHA） mRNA 的表达情况。采用网络药理学分析加味七方胃痛颗粒调控胃癌细胞糖酵解的关键靶点及通路，并通过 Western Blot 检测 PI3K/AKT/HIF-1ɑ 通路相关蛋白表达的变化。结果干预 14 d 后，加味七方胃痛颗粒低、中、高剂量组移植瘤质量均低于对照组（P<0.05），抑瘤率分别为 11.90%、33.61%、52.70%；HE 染色显示加味七方胃痛颗粒低、中、高剂量组较对照组可见不同程度的肿瘤细胞坏死及胞核碎裂；免疫组化结果显示加味七方胃痛颗粒低、中、高剂量组移植瘤中葡萄糖转运蛋白 1（GLUT1），6- 磷酸果糖激酶 1（PFK1）蛋白表达均低于对照组（P<0.05）；微量法和RT-qPCR检测结果显示加味七方胃痛颗粒低、中、高剂量组移植瘤中ATP、乳酸生成量及HK2、LDHA mRNA 表达均低于对照组（P<0.05）；网络药理学分析显示 PI3K/AKT/HIF-1ɑ 信号通路与加味七方胃痛颗粒调控胃癌细胞糖酵解密切相关，Western blot 结果显示加味七方胃痛颗粒低、中、高剂量组移植瘤组织中 p-PI3K/PI3K 蛋白比值及 HIF-1ɑ 蛋白表达均低于对照组（P<0.05）。结论 加味七方胃痛颗粒可以抑制 HGC-27 荷瘤裸鼠移植瘤的生长，其机制与抑制 PI3K/AKT/HIF-1ɑ 通路的活化，降低GLUT1、PFK1、HK2、LDHA 的表达，减少 ATP、乳酸的生成，调控糖酵解有关。

关键词: 胃癌, 加味七方胃痛颗粒, 糖酵解, PI3K/AKT/HIF-1ɑ, 动物实验