Kim Seo Yeon, Yoon Jung-Ho, Jung Da Hyun, Kim Ga Hee, Kim Chul Hoon, Lee Sang Kil
Department of Internal Medicine, Institute of Gastroenterology, Yonsei University College of Medicine, Seoul, Republic of Korea.
Department of Internal Medicine, Graduate School of Medical Science, Brain Korea 21 Project, Yonsei University College of Medicine, Seoul, Republic of Korea.
Front Immunol. 2024 Dec 16;15:1410904. doi: 10.3389/fimmu.2024.1410904. eCollection 2024.
Proton pump inhibitors (PPIs) and potassium-competitive acid blockers (P-CABs) are widely used to manage gastric acid-related disorders by inhibiting hydrochloric acid (HCl) secretion from parietal cells in the stomach. Although PPIs are known to have anti-inflammatory properties beyond their role in inhibiting gastric acid secretion, research on P-CABs is lacking. In this study, we aimed to investigate whether all available P-CABs exhibit anti-inflammatory effects in gastroesophageal reflux-induced esophagitis and to elucidate the underlying mechanisms.
Het-1A cells, normal esophageal epithelial cells, were treated with HCl (pH 4) for 30 min. Esomeprazole, a representative PPI, and three currently marketed P-CABs (vonoprazan, tegoprazan, and fexuprazan) were used for pretreatment. Total RNA sequencing was performed using Het-1A cells pretreated with 1% DMSO or fexuprazan, followed by exposure to HCl. Pyroptosis was measured using lactate dehydrogenase (LDH) release and Annexin V-FITC/PI staining. Western blotting, qRT-PCR, and ELISA were used to determine the expression of the related genes.
Pretreatment with esomeprazole, vonoprazan, tegoprazan, and fexuprazan significantly inhibited the HCl-induced pro-inflammatory cytokines, including IL-6, IL-8, IL-1β, and TNF-α. Fexuprazan and vonoprazan significantly attenuated the HCl-induced pyroptosis rate, as assessed by elevated LDH release and Annexin V-FITC/PI staining, whereas esomeprazole and tegoprazan did not. RNA sequencing revealed that NOD-like receptor (NLR) family pyrin domain-containing 1 (NLRP1) was significantly reduced in Het-1A cells pretreated with fexuprazan compared to those treated with DMSO. Fexuprazan and vonoprazan markedly reduced the HCl-induced transcriptional and translational expression of genes involved in the pyroptosis pathway, including NLRP1, Caspase-1, gasdermin D, and IL-1β. Notably, fexuprazan reduced the HCl-induced increase in pyroptosis and IL-1β using siRNA, even in the presence of NLRP1 knockdown. Fexuprazan, tested on inflammatory THP-1 macrophage cells, significantly reduced NLRP1 expression and inhibited lipopolysaccharide-induced pyroptosis.
Our findings reveal that all p-CABs exhibit anti-inflammatory properties, while fexuprazan inhibits inflammation and pyroptosis of esophageal cells caused by the gastric acid. Therefore, it is presumed to have additional benefits in gastroesophageal reflux disease in addition to suppressing gastric acid secretion.
质子泵抑制剂（PPIs）和钾离子竞争性酸阻滞剂（P-CABs）通过抑制胃壁细胞分泌盐酸（HCl），被广泛用于治疗胃酸相关疾病。尽管已知PPIs除了在抑制胃酸分泌方面发挥作用外，还具有抗炎特性，但关于P-CABs的研究却很缺乏。在本研究中，我们旨在探究所有可用的P-CABs是否在胃食管反流诱导的食管炎中表现出抗炎作用，并阐明其潜在机制。
用HCl（pH 4）处理正常食管上皮细胞Het-1A细胞30分钟。使用代表性的PPI埃索美拉唑和三种目前上市的P-CABs（沃克、替戈拉赞和富马酸伏诺拉生）进行预处理。对用1%二甲基亚砜（DMSO）或富马酸伏诺拉生预处理后再暴露于HCl的Het-1A细胞进行全RNA测序。使用乳酸脱氢酶（LDH）释放和膜联蛋白V-异硫氰酸荧光素/碘化丙啶（Annexin V-FITC/PI）染色来检测细胞焦亡。采用蛋白质免疫印迹法、定量逆转录聚合酶链反应（qRT-PCR）和酶联免疫吸附测定（ELISA）来测定相关基因的表达。
埃索美拉唑、沃克、替戈拉赞和富马酸伏诺拉生预处理可显著抑制HCl诱导的促炎细胞因子，包括白细胞介素-6（IL-6）、白细胞介素-8（IL-8）、白细胞介素-1β（IL-1β）和肿瘤坏死因子-α（TNF-α）。通过升高的LDH释放和Annexin V-FITC/PI染色评估，富马酸伏诺拉生和沃克显著降低了HCl诱导的细胞焦亡率，而埃索美拉唑和替戈拉赞则没有。RNA测序显示，与用DMSO处理的Het-1A细胞相比，用富马酸伏诺拉生预处理的Het-1A细胞中含核苷酸结合寡聚化结构域样受体（NLR）家族含pyrin结构域蛋白1（NLRP1）显著减少。富马酸伏诺拉生和沃克显著降低了HCl诱导的参与细胞焦亡途径的基因的转录和翻译表达，包括NLRP1、半胱天冬酶-1（Caspase-1）、gasdermin D和IL-1β。值得注意的是，即使在NLRP1基因敲低的情况下，富马酸伏诺拉生使用小干扰RNA（siRNA）仍能降低HCl诱导的细胞焦亡增加和IL-1β水平。在炎性THP-1巨噬细胞上进行测试时，富马酸伏诺拉生显著降低NLRP1表达并抑制脂多糖诱导的细胞焦亡。
我们的研究结果表明，所有P-CABs均具有抗炎特性，而富马酸伏诺拉生可抑制胃酸引起的食管细胞炎症和细胞焦亡。因此，推测其除了抑制胃酸分泌外，在胃食管反流病中还具有额外的益处。
