目的 基于转录组学探讨半夏泻心汤（BXD）对大鼠慢性萎缩性胃炎（CAG）的作用机制。方法 采用多因 素联合造模法建立 CAG 大鼠模型，造模总时长 24 周。将 60 只雄性 SD 大鼠随机分为正常组、模型组、阳性 药组（200 mL·kg-1维酶素）及 BXD 低（2.5 g·kg-1 ）、中（5.0g·kg-1 ）、高（10.0 g·kg-1 ）剂量组，每组 10 只。造模成 功后，各给药组大鼠灌胃对应浓度的药物，正常组及模型组大鼠给予等体积蒸馏水灌胃，连续给药 4 周。观察 大鼠体质量等一般状况；采用 HE、阿利新蓝/过碘酸雪夫（AB-PAS）染色法评估大鼠胃组织病理变化并进行病 理学评分。采用试剂盒法检测大鼠胃组织超氧化物歧化酶（SOD）、丙二醛（MDA）、髓过氧化物酶（MPO）、肿瘤 坏死因子 α（TNF-α）、白细胞介素（IL）-1β、IL-6、IL-18 水平。利用转录组学筛选正常组、模型组和给药组 （BXD 高剂量组）大鼠胃组织样本的差异表达基因，并对差异表达基因进行 GO 功能与 KEGG 通路富集分析。 采用 RT-qPCR、Western Blot 法检测大鼠胃组织 NOD 样受体热蛋白结构域相关蛋白 3（NLRP3）、半胱氨酸蛋白 酶 1（Caspase-1）、焦孔素 D（GSDMD）mRNA 及 NLRP3、Cleaved Caspase-1、GSDMD-N 和 Cleaved IL-1β 蛋白 表达水平。结果 （1）与正常组比较，模型组大鼠胃窦部及胃体底部黏膜上皮出现明显萎缩、变薄，腺体数量 减少，有淋巴细胞浸润、淋巴滤泡形成、肠上皮化生以及不同程度的异型增生；胃黏膜上皮细胞 AB-PAS 染色 可见明显的蓝染灶，兼见蓝紫染色灶，肠上皮化生病灶范围较广。经药物干预后，BXD 低剂量组大鼠胃黏膜 病理变化相较于模型组有好转，但仍有一些腺腔扩张和淋巴细胞浸润，胃黏膜上皮细胞 AB-PAS 染色仍可见部 分蓝染病灶；阳性药组和 BXD 中、高剂量组大鼠胃黏膜病变的严重程度明显减轻，无明显的肠上皮化生， AB-PAS 染色的胃黏膜细胞以红染为主，少见蓝染病灶。（2）与正常组比较，模型组大鼠体质量与胃组织 SOD 含量降低（P＜0.01），胃组织病理学评分及 MPO、MDA、IL-6、IL-18、TNF-α、IL-1β 含量升高（P＜0.01）。 与模型组比较，各给药组大鼠体质量及胃组织 SOD 含量升高（P＜0.05，P＜0.01），各给药组大鼠胃组织中 IL-6、 IL-18、IL-1β 含量及 BXD 高剂量组、阳性药组大鼠胃组织病理学评分和 TNF-α、MPO、MDA 含量降低（P＜ 0.05，P＜0.01），BXD 低、中剂量组大鼠胃组织病理学评分、TNF-α 含量及 BXD 低剂量组大鼠 MPO 和 MDA 含量有降低的趋势，但差异无统计学意义（P＞0.05）。（3）转录组学结果显示，模型组 vs. 正常组有差异表达基因 1 032 个（上调 632 个、下调 400 个）；给药组 vs. 模型组有差异表达基因 757 个（上调 246 个、下调 511 个）。经 BXD 干预后，554 个交集差异表达基因出现回调，其中给药组中 379 个差异表达基因较模型组降低，175 个差异表 达基因表达升高。554 个回调差异表达基因 KEGG 显著富集到其它类型的 O-聚糖生物合成、PI3K-Akt 信号通 路、NOD 样受体通路和钙信号通路等，且聚类分析得到 NOD 样受体通路中 NLRP3、Caspase-1 及 GSDMD 等 基因的差异表达明显（P＜0.05）。（4）与正常组比较，模型组大鼠胃组织中 NLRP3、Caspase-1、GSDMD mRNA 及 NLRP3、Cleaved Caspase-1、GSDMD-N 和 Cleaved IL-1β 蛋白表达水平升高（P＜0.01）。与模型组比较， BXD 各给药组大鼠胃组织中 Cleaved Caspase-1、GSDMD-N、Cleaved IL-1β 蛋白及 BXD 中、高剂量组大鼠胃 组织中 NLRP3、Caspase-1、GSDMD mRNA 及 NLRP3 蛋白表达水平明显降低（P＜0.05，P＜0.01），BXD 低剂 量组大鼠胃组织中 NLRP3、Caspase-1、GSDMD mRNA 及 NLRP3 蛋白有降低趋势，但差异无统计学意义（P＞0.05）。结论 BXD 对 CAG 大鼠的病理状况具有改善作用，其可能通过调控 NLRP3/Caspase-1/GSDMD 信号通 路，减轻 GAG 大鼠氧化应激、炎症反应及减少细胞焦亡的发生，保护胃黏膜，从而发挥治疗 CAG 的作用。

Objective To explore the mechanism of Banxia Xiexin Decoction （BXD） in ameliorating chronic atrophic gastritis （CAG） based on transcriptomics. Methods A CAG rat model was established using a multifactorial combined modeling method over a total period of 24 weeks. Sixty male SD rats were randomly divided into six groups （n=10）：a normal control group，a model group，a positive drug group （vitacoenzyme，200 mL·kg⁻¹），low （2.5 g·kg⁻¹），medium- （5.0 g·kg⁻¹），and high- （10.0 g·kg⁻¹）dose BXD groups. After successful modeling，rats in the treatment groups were administered the corresponding drugs via gavage，while rats in the normal and model groups received an equal volume of distilled water for 4 consecutive weeks. General conditions such as body mass were observed. Pathological changes in gastric tissue were evaluated by hematoxylin-eosin （HE） and Alcian blue-periodic acid Schiff （AB-PAS） staining， and pathological scores were assigned. Levels of superoxide dismutase （SOD）， malondialdehyde （MDA）， myeloperoxidase （MPO），tumor necrosis factor-alpha （TNF-α），interleukin （IL）-1β，IL-6，and IL-18 in gastric tissue were detected using assay kits. Transcriptomic analysis was performed to screen differentially expressed genes （DEGs） in gastric tissue samples from the normal，model，and treatment （BXD high-dose） groups. GO functional and KEGG pathway enrichment analyses were conducted on the DEGs. mRNA expression levels of NOD-like receptor thermal protein domain associated protein 3（NLRP3）， Caspase-1， and gasdermin D （GSDMD）， and protein expression levels of NLRP3，Cleaved Caspase-1，GSDMD-N，and Cleaved IL-1β in gastric tissue were measured by RT-qPCR and Western Blot，respectively. Results （1） Compared with the normal group，the model group showed marked atrophy and thinning of the mucosal epithelium in the gastric antrum and corpus fundus，with a reduction in gland number. Lymphocyte infiltration， lymphoid follicle formation， intestinal metaplasia， and varying degrees of dysplasia were observed. AB-PAS staining of gastric mucosal epithelial cells revealed distinct blue-stained foci and blue-purple foci， with extensive intestinal metaplasia lesions. After drug intervention， pathological changes in the gastric mucosa of the low-dose BXD group showed improvement compared to the model group， but some glandular dilatation and lymphocyte infiltration remained， and AB-PAS staining still showed partial blue-stained foci. The severity of gastric mucosal lesions was significantly alleviated in the positive drug group and the medium- and high-dose BXD groups， with no obvious intestinal metaplasia. AB-PAS staining of gastric mucosal cells in these groups was predominantly red， with rare blue-stained foci.（2） Compared with the normal group， the model group exhibited decreased body mass and gastric SOD content （P＜0.01） and increased gastric pathological scores and levels of MPO， MDA，IL-6，IL-18，TNF-α，and IL-1β（P＜0.01）. Compared with the model group，all treatment groups showed increased body mass and gastric SOD content （P＜0.05，P＜0.01）. All treatment groups showed decreased levels of gastric IL-6， IL-18， and IL-1β （P＜0.05， P＜0.01）. The high-dose BXD and positive drug groups showed decreased gastric pathological scores and levels of TNF- α， MPO， and MDA （P＜0.05， P＜0.01）. Although decreasing trends were observed for pathological scores and TNF- α levels in the low- and medium-dose BXD groups and for MPO and MDA levels in the low-dose BXD group，the differences were not statistically significant （P＞0.05）.（3） Transcriptomic results revealed 1 032 DEGs （632 up-regulated，400 down-regulated） in the model group vs. the normal group，and 757 DEGs （246 up-regulated，511 down-regulated） in the treatment group vs. the model group. After BXD intervention， 554 intersecting DEGs showed a reversion trend： 379 DEGs were down-regulated and 175 were up-regulated in the treatment group compared to the model group. These 554 reversed DEGs. KEGG pathway enrichment analysis showed significant enrichment in pathways including other types of O-glycan biosynthesis，PI3KAkt signaling pathway， NOD-like receptor signaling pathway， and calcium signaling pathway. Cluster analysis indicated significant differential expression of genes in the NOD-like receptor pathway，such as NLRP3，Caspase-1，and GSDMD （P＜0.05）.（4）Compared with the normal group， the model group showed increased mRNA expression of NLRP3，Caspase-1，and GSDMD，and increased protein expression of NLRP3，Cleaved Caspase-1，GSDMD-N， and Cleaved IL-1β in gastric tissue （P＜0.01）. Compared with the model group，all BXD treatment groups showed significantly decreased protein expression of Cleaved Caspase-1， GSDMD-N， and Cleaved IL-1β（P＜0.05， P＜ 0.01）. The medium- and high-dose BXD groups showed significantly decreased mRNA expression of NLRP3，Caspase-1， and GSDMD and protein expression of NLRP3（P＜0.05， P＜0.01）. Although decreasing trends were observed for NLRP3， Caspase-1， GSDMD mRNA and NLRP3 protein in the low-dose BXD group， the differences were not statistically significant （P＞0.05）. Conclusion BXD ameliorates pathological conditions in CAG rats. Its therapeutic effect may be achieved by regulating the NLRP3/Caspase-1/GSDMD signaling pathway， thereby reducing oxidative stress，inflammatory response，and pyroptosis in CAG rats，and protecting the gastric mucosa.

R285.5

国家自然科学基金面上项目（82274446，81774183）。