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目的:本研究旨在探讨加味柴芍六君方对胃癌上皮间质转化(EMT)过程中白细胞介素-8(IL-8)的调节机制，为胃癌治疗提供新的策略。方法:通过MTT法检测加味柴芍六君方对人胃黏膜细胞(GES-1)的毒性及对胃癌细胞(HCG-27)增殖的影响；通过划痕试验和Transwell侵袭试验评估该方对细胞迁移和侵袭能力的抑制作用；利用TGF-β1 10μg/L诱导EMT模型，并通过慢病毒转染构建IL-8过表达细胞模型；采用ELISA检测癌胚抗原(CEA)、胃泌素(G-17)和IL-8含量，qRT-PCR和Western blot法检测EMT标志物(Ecadherin、Ncadherin、Vimentin、Twist、Snail)及Il8的mRNA和蛋白表达；免疫荧光法观察IL-8蛋白定位。结果:相较于空白对照组，加味柴芍六君方400μg/mL组HCG-27细胞的增殖、迁移和侵袭降低(P<0.01),TGF-β1组、IL-8组，加味柴芍六君方200、400μg/mL组细胞CEA、G-17和IL-8含量显著升高(P<0.01),过表达IL-8组细胞CEA、G-17、IL-8含量显著升高(P<0.01)。与过表达IL-8模型对照组比较，加味柴芍六君方200、400μg/mL组CEA、G-17和IL-8的含量明显降低(P<0.05或P<0.01);加味柴芍六君方400μg/mL+过表达组Ncadherin、Vimentin、Twist、Snail、IL-8蛋白和mRNA表达下调，Ecadherin上调(P<0.05或P<0.01);TGF-β1组细胞Twist mRNA表达上调，Il8、EcadherinmRNA表达下调(P<0.05或P<0.01)。与TGF-β1组比较，加味柴芍六君方+TGF-β1组细胞Vimentin、Twist、SnailmRNA表达下调，Ecadherin、Il8 mRNA表达上调(P<0.05或P<0.01)。免疫荧光结果表明，相较于空白对照组，过表达IL-8组和TGF-β1组的荧光强度较大，而加味柴芍六君方组减弱细胞IL-8的荧光强度。结论:加味柴芍六君方通过调控IL-8及其下游信号通路，显著抑制了胃癌细胞的EMT过程。
Abstract：Objective:To investigate the regulatory mechanism of modified Chaishao Liujun Prescription(柴芍六君方) on interleukin-8(IL-8) during epithelial-mesenchymal transition(EMT) in gastric cancer, and to provide new strategies for its treatment. Methods:The cytotoxicity of modified Chaishao Liujun Prescription on human gastric mucosal cells(GES-1) and its effects on the proliferation of gastric cancer cells(HCG-27) were detected by MTT assay. Scratch wound healing and Transwell invasion assays were used to evaluate its inhibitory effects on cell migration and invasion. An EMT model was induced by TGF-β1(10 μg/L),and an IL-8 overexpression model was constructed by lentiviral transfection. ELISA was used to measure the secretion levels of carcinoembryonic antigen(CEA),gastrin-17(G-17),and IL-8. The proteins and mRNA expression changes of EMT markers(E-cadherin, N-cadherin, Vimentin, Twist, Snail) and IL-8 were analyzed by Western blot and qRT-PCR. Immunofluorescence was used to observe the localization of IL-8 protein. Results:Compared with the blank control group, modified Chaishao Liujun Prescription(400 μg/mL) grop significantly inhibited the proliferation, migration, and invasion(P<0.01) of HCG-27 cells, the TGF-β1 group, the IL-8 group, and modified Chaishao Liujun Prescription(200, 400 μg/mL) groups exhibited significantly elevated levels of CEA,G-17,and IL-8(P<0.01) of cells, and the IL-8 overexpression group exhibited significantly elevated levels of CEA,G-17,and IL-8(P<0.01) of cells. Compared with the IL-8 overexpression model control group, modified Chaishao Liujun Prescription(200, 400 μg/mL) groups exhibited significantly reduced levels of CEA,G-17,and IL-8(P<0.05 or P<0.01) of cells, modified Chaishao Liujun Prescription(400 μg/mL) + overexpression group exhibited significantly downregulated the protein and mRNA expression of N-cadherin, Vimentin, Twist, Snail, and IL-8, while upregulating E-cadherin(P<0.05 or P<0.01) of cells, and the TGF-β1 group showed upregulated the mRNA expression of Twist, while downregulating Il8, Ecadherin(P<0.05 or P<0.01) of cells. Compared with the TGF-β1 group, modified Chaishao Liujun Prescription + TGF-β1 group showed downregulated the mRNA expression of Vimentin,Twist,and Snail,while upregulating E-cadherin and IL-8(P<0.05 or P<0.01) of cells. Immunofluorescence results indicated that, compared with the blank control group, the IL-8 overexpression group and the TGF-β1 group showed stronger fluorescence intensity, whereas the modified Chaishao Liujun Prescription group attenuated IL-8 fluorescence signal of cells. Conclusion: By regulating IL-8 and its downstream signaling pathways, the modified Chaishao Liujun Prescription significantly inhibited EMT in gastric cancer cells.
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基本信息:
DOI：10.13412/j.cnki.zyyl.20250929.011
中图分类号:R285
引用信息:
[1]蒋敏玲,文辉,何武,等.加味柴芍六君方抑制IL-8介导的胃癌上皮间质转化的机制研究[J].中药药理与临床,2026,42(06):2-10.DOI:10.13412/j.cnki.zyyl.20250929.011.
基金信息:
广西中医药管理局自筹经费科研课题(编号：GXZYZ20210091)
2025-09-30
2025-09-30
2025-09-30