中药新药与临床药理2025，Vol.36Issue(12)：2051-2061,11.DOI:10.19378/j.issn.1003-9783.2025.12.005
健脾化瘀解毒方通过影响JAK2/STAT3信号通路调节ROS抑制胃癌前病变细胞恶性转化
Jianpi Huayu Jiedu Formula Suppresses Malignant Transformation of Gastric Precancerous Lesions by Regulating ROS via the JAK2/STAT3 Signaling Pathway
摘要
Abstract
Objective To investigate the mechanism of Jianpi Huayu Jiedu Formula(JPJD)in the treatment of gastric precancerous lesions(GPL)based on network pharmacology.Methods(1)The active components and potential targets of JPJD were analyzed using the TCM Systems Pharmacology Database and Analysis Platform(TCMSP),the Human Gene Database(GeneCards),and the Human Disease Database(MalaCards).A network of JPJD active components and GPL targets was constructed,and core targets were subjected to Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis.(2)A GPL cell model was established by inducing the human gastric mucosal epithelial cell line GES-1 with N-methyl-N-nitrosourea(MNU).Different concentrations of JPJD were applied to the GPL cells,and cell viability was assessed using the CCK-8 assay.The experiment was divided into the following groups:blank control group,MNU group(model group),MNU+low-concentration JPJD group,MNU+medium-concentration JPJD group,and MNU+high-concentration JPJD group.The mRNA expression levels of GPL-related markers(P53,MUC2,TFF2)were detected by RT-PCR.Cell migration ability was evaluated using the Transwell assay.Cell proliferation capacity was assessed by the plate clone formation assay.Intracellular ROS levels were measured using a ROS fluorescent probe.The protein expression levels of IL-6,JAK2,STAT3,and p-JAK2 were determined by Western Blot.Results(1)Screening identified 20 active components from Astragali Radix,8 from Gecko,8 from Hedyotis Diffusae Herba,15 from Poria,8 from Pseudostellariae Radix,7 from Atractylodis Macrocephalae Rhizoma,3 from Curcumae Rhizoma,8 from Hericium erinaceus,and 8 from Notoginseng Radix et Rhizoma.The intersection of active component targets and disease-related targets,obtained via the Venny 2.1 platform,yielded 121 potential therapeutic targets of JPJD for GPL.The core targets identified for JPJD in treating GPL were TNF,VEGFA,AKT1,IL-6,and STAT3.(2)CCK-8 and RT-PCR results indicated that an MNU concentration of 100 μmol·mL-1 did not affect cell viability,while the expression levels of GPL-related markers showed varying degrees of alteration(P＜0.01,P＜0.001).(3)CCK-8 results suggested that JPJD at intervention concentrations of 1,2,and 3 mg·mL-1 did not affect cell viability.(4)The Transwell assay showed that compared with the blank control group,the number of migrated cells in the model group increased(P＜0.05);compared with the model group,the number of migrated cells in medium-and high-concentration JPJD treatment groups decreased in a concentration-dependent manner(P＜0.001).(5)The plate clone formation assay showed that compared with the blank control group,the number of cell clones in the model group significantly increased(P＜0.05);compared with the model group,the number of cell clones decreased in medium-and high-concentration JPJD treatment groups(P＜0.001).(6)ROS detection results indicated that compared with the blank control group,ROS levels were significantly increased in the model group(P＜0.001);compared with the model group,ROS levels were decreased in the JPJD treatment groups(P＜0.05,P＜0.01).(7)Western Blot results showed that compared with the blank control group,the protein expression levels of IL-6,JAK2,STAT3,and p-JAK2 were elevated in the model group(P＜0.05,P＜0.01,P＜0.001);compared with the model group,the protein expression levels of IL-6,JAK2,STAT3,and p-JAK2 were decreased in medium-and high-concentration JPJD treatment groups(P＜0.001).Conclusion Jianpi Huayu Jiedu Formula may inhibit the progression of gastric precancerous lesions by suppressing ROS accumulation and consequently inhibiting the activation of the JAK2/STAT3 signaling pathway.关键词
健脾化瘀解毒方/胃癌前病变/JAK2/STAT3信号通路/活性氧/细胞恶化/人胃黏膜上皮细胞GES-1Key words
Jianpi Huayu Jiedu Formula/gastric precancerous lesions/JAK2/STAT3/reactive oxygen species/malignant transformation/human gastric mucosal epithelial cell line GES-1分类
医药卫生引用本文复制引用
张思佳,方崇锴,何佩瑶,刘伟,谢梓淳,潘华峰..健脾化瘀解毒方通过影响JAK2/STAT3信号通路调节ROS抑制胃癌前病变细胞恶性转化[J].中药新药与临床药理,2025,36(12):2051-2061,11.基金项目
广东省普通高校创新团队项目(2023KCXTD080) （2023KCXTD080）
国家自然科学基金青年科学基金项目(82405319) （82405319）
中国博士后科学基金面上项目(2024M750666). （2024M750666）
