文献类型：期刊文献

中文题名：基于Jurkat-胃癌细胞共培养体系探究归芪白术方小分子治疗胃癌的作用机制

英文题名：Mechanism of small molecules derived from the Guiqi Baizhu formula in the treatment of gastric cancer based on a Jurkat-gastric cancer cell co-culture system

作者：马欢欢[1,2];孔凡明[1,2];何振宇[1,2];庞月苓[1,2];房明[1,2];李佳蔚[1,2];刘永琦[1,2,3];李玲[1,2]

第一作者：马欢欢

机构：[1]甘肃中医药大学基础医学院中西医结合基础专业,兰州730000;[2]甘肃中医药大学甘肃省高校重大疾病分子医学与中医药防治研究重点实验室,兰州730000;[3]甘肃中医药大学敦煌医学与转化教育部重点实验室,兰州730000

第一机构：甘肃中医药大学基础医学院（敦煌医学研究所）|甘肃中医药大学中西医结合学院

年份：2026

卷号：42

期号：5

起止页码：331

中文期刊名：免疫学杂志

外文期刊名：Immunological Journal

基金：国家自然科学基金(82205318);甘肃中医药大学引进人才科研启动基金项目(2023YJRC-08);甘肃省联合科研基金重大项目(24JRRA875)。

语种：中文

中文关键词：胃癌;共培养;Jurkat细胞;小分子;归芪白术方;小鼠

外文关键词：gastric cancer;co-culture;Jurkat cells;small molecules;Guiqi Baizhu formula;mice

摘要：目的探讨归芪白术方小分子通过健脾化瘀法治疗胃癌的作用机制。方法构建Jurkat-胃癌细胞共培养体系,分为共培养组(未予任何处理)、(Z)-Ligustilide(LIG)组、LIG+Glycycoumarin(GLY)组,后两组分别使用靶向血管内皮生长因子-A(VEGF-A)的小分子LIG(10μmol/L)、靶向程序性死亡受体配体1(PD-L1)的小分子GLY(10μmol/L)单独及联合干预共培养体系,干预24 h。选取56只健康的雄性615小鼠,随机分为7组,每组8只。分别为空白对照组(正常615小鼠)、模型组(胃癌荷瘤小鼠)、顺铂组、顺铂+归芪白术方组、顺铂+LIG组、顺铂+GLY组、顺铂+LIG+GLY组。空白对照组不干预,模型组灌胃0.9%氯化钠溶液,顺铂(浓度为5 mg/kg)每4天腹腔注射一次,LIG(浓度为40 mg/kg)每2天腹腔注射1次,GLY(浓度为20 mg/kg)每天腹腔注射一次,干预14 d。流式细胞术、高内涵实时监测中药小分子配伍干预共培养体系后胃癌细胞的状态及凋亡情况,免疫组织化学、酶联免疫吸附试验分别检测肿瘤组织中蛋白表达变化及小鼠脾脏和血清中γ干扰素(IFN-γ)、白细胞介素(IL)-2含量。结果与共培养组比较,LIG组、LIG+GLY组共培养模型中胃癌细胞逐渐皱缩、凋亡,细胞凋亡率均升高(P<0.05)。与模型组比较,各给药组肿瘤体积、质量及VEGF-A、Ki-67、程序性死亡蛋白-1(PD-1)、PD-L1蛋白表达水平均降低(P<0.05);与空白组比较,模型组小鼠脾脏和血清中IL-2、IFN-γ含量均降低(P<0.05);与模型组比较,顺铂组脾脏和血清中IL-2、IFN-γ含量均降低,而顺铂+归芪白术方组、顺铂+GLY组和顺铂+LIG+GLY组干预后,脾脏和血清中IL-2和IFN-γ浓度均升高(P<0.05)。结论LIG联合GLY可通过健脾益气、活血化瘀抑制小鼠肿瘤生长并改善肿瘤微环境中T淋巴细胞免疫活性,发挥抗肿瘤效果。
Objective To investigate the mechanism of small molecules derived from the Guiqi Baizhu formula in the treatment of gastric cancer through the method of strengthening the spleen and resolving blood stasis.Methods A Jurkat-gastric cancer cell co-culture system was established,and the cells were divided into a co-culture group(no treatment was given),a(Z)-Ligustilide(LIG)group,and a LIG+Glycycoumarin(GLY)group.The latter two groups were treated with the small molecule LIG(10μmol/L)targeting vascular endothelial growth factor A(VEGF-A),and the small molecule GLY(10μmol/L)targeting programmed cell death ligand 1(PD-L1),either alone or in combination,in the co-culture system for 24 h.Fifty-six healthy male 615 mice were selected and randomly divided into 7 groups,with 8 mice in each group:blank control group(normal 615 mice),model group(gastric cancer-bearing mice),Cisplatin group,Cisplatin+Guiqi Baizhu formula group,Cisplatin+LIG group,Cisplatin+GLY group,and Cisplatin+LIG+GLY group.The blank control group received no intervention.The model group was administered 0.9%Sodium Chloride solution by gavage.Cisplatin was administered at a concentration of 5 mg/kg via intraperitoneal injection every 4 d,LIG via intraperitoneal injection at 40 mg/kg every 2 d,and GLY via intraperitoneal injection at 20 mg/kg daily,for a total intervention period of 14 d.Flow cytometry and highcontent real-time monitoring were used to assess the status and apoptosis of gastric cancer cells following intervention with traditional Chinese medicine small-molecule combinations in the co-culture system.Immunohistochemistry and enzymelinked immunosorbent assays(ELISA)were used to detect changes in protein expression in tumor tissues and the levels of interferon-gamma(IFN-γ)and interleukin(IL)-2 in mouse spleen and serum,respectively.Results Compared with the coculture group,gastric cancer cells in the LIG group and the LIG+GLY group gradually shrank and underwent apoptosis,and the apoptosis rates were significantly higher(P<0.05).Compared with the model group,tumor volume,mass,and expression levels of VEGF-A,Ki-67,programmed death protein-1(PD-1),and PD-L1 were all reduced in all treatment groups(P<0.05).Compared with the control group,levels of IL-2 and IFN-γin the spleen and serum of mice in the model group were both reduced(P<0.05).Compared with the model group,IL-2 and IFN-γlevels in the spleen and serum were both reduced in the Cisplatin group;however,after intervention in Cisplatin+Guiqi Baizhu formula group,Cisplatin+GLY group,and Cisplatin+LIG+GLY group,IL-2 and IFN-γlevels in the spleen and serum were both elevated(P<0.05).Conclusion The combination of LIG and GLY exerts antitumor effects by inhibiting tumor growth in mice and improving T-lymphocyte immune activity in the tumor microenvironment through spleen-strengthening,qi-nourishing,and blood-activating,stasis-resolving actions.