Abstract: Background　Chronic atrophic gastritis（CAG） is an important pathological stage in the progression from gastric mucosal inflammation to cancer. The TLR4 and Sonic Hedgehog（Shh） pathways play key roles in gastric mucosal injury and repair，while the NLRP3/Caspase-1 pathway is also involved in the inflammatory response and pyroptosis of gastric mucosal epithelial cells. The compound gastric inflammation mixture（CGM） has shown significant efficacy in the treatment of CAG， but its specific mechanism of action remains unclear. Objective　To observe the effects of CGM-containing serum on pyroptosis of gastric mucosal epithelial cells by modulating the NLRP3/Caspase-1 pathway through the crosstalk between TLR4 and Shh signaling pathways，thereby exploring the potential mechanisms by which CGM treats CAG. Methods　Sixteen Wistar rats were randomly divided into a blank group and a CGM group，with 8 rats in each group. After continuous intragastric administration of CGM suspension for 10 days，serum was collected from the CGM group. The CCK-8 method was used to screen the CGM-containing serum，determining the optimal intervention concentration to be 10% and the optimal intervention time to be 48 hours. An ethanol-induced cell model was established to mimic CAG in human gastric mucosal epithelial cells. Cells that successfully underwent modeling were divided into blank，model，CGM，TLR4 inhibitor，TLR4 inhibitor+CGM，TLR4 agonist，TLR4 agonist+CGM，Shh inhibitor，Shh inhibitor+CGM，Shh agonist，and Shh agonist+CGM groups. The expression levels of TLR4，NF-κB，Shh，NLRP3，and Caspase-1 mRNA and protein were detected using real-time quantitative PCR（RT-qPCR） and Western blotting，while the levels of interleukin（IL）-1β，IL-6，and IL-18 in cell supernatants were measured using enzyme-linked immunosorbent assay（ELISA）. Results　Compared to the blank group，the model group exhibited increased mRNA and protein levels of TLR4，NF-κB，NLRP3，and Caspase-1（P<0.05），along with decreased mRNA and protein levels of Shh（P<0.05）. The levels of IL-1β，IL-6，and IL-18 in the cell supernatants were significantly elevated （P<0.05）. Compared to the model group，the CGM group，TLR4 inhibitor group，and Shh agonist group showed decreased mRNA levels of TLR4，NF-κB，NLRP3，and Caspase-1，while Shh mRNA levels were elevated（P<0.05）. The CGM group also demonstrated decreased protein expression levels of NF-κB，NLRP3，and Caspase-1（P<0.05）. The TLR4 inhibitor group showed reduced protein expression of TLR4，NF-κB，NLRP3，and Caspase-1（P<0.05） and increased Shh protein expression（P<0.05）. In the TLR4 agonist group，TLR4 protein expression was elevated（P<0.05）. The Shh inhibitor group exhibited increased protein levels of TLR4 and NF-κB（P<0.05）. The Shh agonist group showed decreased protein expression of NF-κB，NLRP3，and Caspase-1，alongside increased Shh protein expression（P<0.05）. The levels of IL-1β，IL-6，and IL-18 in the cell supernatants were significantly reduced in the CGM group（P<0.05）. The TLR4 inhibitor group and Shh agonist group had decreased IL-1β levels in their cell supernatants（P<0.05），while the TLR4 agonist group and Shh inhibitor group showed increased IL-1β levels（P<0.05）. Compared to the CGM group，the TLR4 inhibitor + CGM group exhibited decreased mRNA levels of TLR4 and Caspase-1（P<0.05），along with reduced mRNA and protein expression levels of NF-κB and NLRP3（P<0.05），and increased mRNA and protein expression levels of Shh（P<0.05）. In the TLR4 agonist+CGM group，mRNA levels of TLR4 and NF-κB were elevated（P<0.05），while Shh mRNA expression decreased（P<0.05）. The Shh inhibitor+CGM group showed decreased Shh mRNA levels（P<0.05）. In the Shh agonist+CGM group，mRNA and protein expression levels of TLR4，NF-κB，and Caspase-1 were reduced（P<0.05），while Shh mRNA and protein expression levels were increased（P<0.05）. Both the TLR4 agonist + CGM group and Shh inhibitor + CGM group demonstrated elevated serum levels of IL-1β，IL-6，and IL-18（P<0.05）. Conclusion　CGM-containing serum may regulate the NLRP3/Caspase-1 pathway and mediate the pyroptosis process of gastric epithelial cells by influencing the crosstalk between the TLR4 and Shh pathways，thereby improving the inflammatory-cancer transformation process in ethanol-induced GES-1 cells and exerting therapeutic effects on CAG.

Key words: Chronic atrophic gastritis, Compound gastritis mixture, Crosstalk, NLRP3/Caspase-1 pathway, Pyroptosis

摘要： 背景　慢性萎缩性胃炎（CAG）是胃黏膜炎-癌转化的重要病理阶段，TLR4与Sonic Hedgehog（Shh）通路在胃黏膜损伤与修复中起关键作用，NLRP3/Caspase-1通路亦参与胃黏膜上皮细胞的炎症反应和焦亡过程。复方胃炎合剂（CGM）在CAG治疗中效果显著，但其具体作用机制尚不清楚。目的　观察CGM含药血清通过TLR4与Shh信号通路串话调控NLRP3/Caspase-1通路对胃黏膜上皮细胞焦亡的影响，探讨CGM治疗CAG可能的作用机制。方法　将16只Wistar大鼠随机分为空白组和CGM组，每组各8只。CGM组以CGM混悬液连续灌胃10 d后采集血清。采用CCK-8法筛选CGM含药血清，测得最佳干预浓度为10%，最佳干预时间为48 h。采用乙醇诱导人胃黏膜上皮细胞建立CAG细胞模型。将造模成功细胞分为空白组、模型组、CGM组、TLR4抑制剂组、TLR4抑制剂+CGM组、TLR4激动剂组、TLR4激动剂+CGM组、Shh抑制剂组、Shh抑制剂+CGM组、Shh激动剂组、Shh激动剂+CGM组。采用实时荧光定量PCR（RT-qPCR）和Western blotting法分别检测细胞TLR4、NF-κB、Shh、NLRP3、Caspase-1 mRNA与蛋白表达，酶联免疫吸附试验（ELISA）法检测细胞上清中白介素（IL）-1β、IL-6、IL-18水平。结果　与空白组比较，模型组细胞TLR4、NF-κB、NLRP3、Caspase-1 mRNA与蛋白表达均升高（P<0.05），Shh mRNA与蛋白表达下降（P<0.05），细胞上清中IL-1β、IL-6、IL-18水平均升高（P<0.05）。与模型组相比，CGM组、TLR4抑制组与Shh激动组TLR4、NF-κB、NLRP3、Caspase-1 mRNA水平均下降，Shh mRNA水平升高（P<0.05）；CGM组NF-κB、NLRP3、Caspase-1蛋白表达均下降（P<0.05）；TLR4抑制组TLR4、NF-κB、NLRP3、Caspase-1蛋白表达均下降（P<0.05），Shh蛋白表达上升（P<0.05）；TLR4激动剂组TLR4蛋白表达升高（P<0.05）；Shh抑制剂组TLR4、NF-κB蛋白表达升高（P<0.05）；Shh激动剂组NF-κB、NLRP3、Caspase-1蛋白表达均下降，Shh蛋白表达升高（P<0.05）；CGM组细胞上清中IL-1β、IL-6、IL-18水平均降低（P<0.05）；TLR4抑制剂组与Shh激动剂组细胞上清中IL-1β水平均下降（P<0.05），TLR4激动剂组、Shh抑制剂组IL-1β水平升高（P<0.05）。与CGM组相比，TLR4抑制剂+CGM组TLR4、Caspase-1 mRNA水平下降（P<0.05），NF-κB、NLRP3 mRNA与蛋白表达下降（P<0.05），Shh mRNA与蛋白表达升高（P<0.05）；TLR4激动剂+CGM组TLR4、NF-κB mRNA水平升高（P<0.05），Shh mRNA表达下降（P<0.05）；Shh抑制剂+CGM组Shh mRNA水平下降（P<0.05）；Shh激动剂+CGM组TLR4、NF-κB、Caspase-1 mRNA与蛋白表达下降（P<0.05），Shh mRNA与蛋白表达升高（P<0.05）；TLR4激动剂+CGM组和Shh抑制剂+CGM组血清IL-1β、IL-6与IL-18均升高（P<0.05）。结论　CGM含药血清可能通过影响TLR4与Shh通路串话调控NLRP3/Caspase-1通路介导胃上皮细胞焦亡过程，改善乙醇诱导GES-1细胞的炎-癌转化进程，从而发挥治疗CAG的作用。

关键词: 慢性萎缩性胃炎, 复方胃炎合剂, 串话, NLRP3/Caspase-1 通路, 细胞焦亡