目的 基于网络药理学探讨健脾化瘀解毒方（JPJD）治疗胃癌前病变的作用机制。方法 （1）利用中药系 统药理学数据库与分析平台、人类基因数据库、人类疾病相关数据库等分析健脾化瘀解毒方组方活性成分及潜 在作用靶点，构建组方活性成分与胃癌前疾病靶点网络，并对核心靶点进行基因本体（GO）及京都基因与基因 组百科全书（KEGG）通路富集分析。（2）使用甲基亚硝基脲（MNU）诱导人胃黏膜上皮细胞 GES-1 建立胃癌前病变 （GPL）细胞模型，将不同浓度的健脾化瘀解毒方作用于胃癌前病变细胞，采用 CCK-8 法评估细胞活力；将实验 分为空白组、MNU 组（模型组）、MNU+健脾化瘀解毒方（低浓度组）、MNU+健脾化瘀解毒方（中浓度组）、 MNU+健脾化瘀解毒方组（高浓度组）；RT-PCR 法检测胃癌前病变相关标志物 P53、MUC2、TFF2 的 mRNA 表 达情况；Transwell 实验检测胃癌前病变细胞迁移能力；平板克隆实验检测细胞增殖能力；ROS 探针检测胃癌 前病变细胞的 ROS 水平；Western Blot 法检测胃癌前病变细胞的 IL-6、JAK2、STAT3、p-JAK2 蛋白表达。 结果 （1）筛选出活性成分黄芪 20 个、守宫 8 个、白花蛇舌草 8 个、茯苓 15 个、太子参 8 个、白术 7 个、莪 术 3 个、猴头菇 8 个、三七 8 个。将活性成分作用靶点与疾病相关靶点经 Venny 2.1 平台取交集，共得到健脾 化瘀解毒方治疗胃癌前病变的潜在作用靶点 121 个。筛选得到健脾化瘀解毒方治疗胃癌前病变的核心靶点： TNF、VEGFA、AKT1、IL-6、STAT3。（2）CCK-8 及 RT-PCR 检测结果显示，MNU 浓度为 100 μmol·mL -1 时不 会影响细胞活性，且胃癌前病变的相关标志物有不同程度变化（P＜0.01，P＜0.001）。（3）CCK-8 法检测结果提 示，健脾化瘀解毒方在干预浓度为 1、2、3 mg·mL -1 时，不会对细胞活力产生影响。（4）Transwell 实验表明，与 空白组比较，模型组细胞迁移数目增多（P＜0.05）；与模型组比较，健脾化瘀解毒方中、高剂量组细胞迁移数 目呈浓度依赖性减少（P＜0.001）。（5）平板克隆实验表明，与空白组比较，模型组细胞克隆数量明显增多（P＜ 0.05）；与模型组比较，健脾化瘀解毒方中、高剂量组细胞克隆数量下降（P＜0.001）。（6）ROS 法检测结果提示， 与空白组比较，模型组 ROS 含量显著增多（P＜0.001）；与模型组比较，给药组 ROS 含量减少（P＜0.05，P＜ 0.01）。（7）Western Blot 结果提示，与空白组比较，模型组 IL-6、JAK2、STAT3、p-JAK2 蛋白表达水平升高 （P＜0.05，P＜0.01，P＜0.001）；与模型组比较，健脾化瘀解毒方中、高剂量组 IL-6、JAK2、STAT3、p-JAK2 蛋白表达水平降低（P＜0.001）。结论 健脾化瘀解毒方可能通过抑制 ROS 的积累从而抑制 JAK2/STAT3 信号通 路的激活以阻滞胃癌前病变的发展。

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.

R285.5

广东省普通高校创新团队项目（2023KCXTD080)；国家自然科学基金青年科学基金项目（82405319）；中国博士后科学基金面上项目 （2024M750666）。