Acute pancreatitis (AP) is an inflammation-associated disorder in the digestive system. Ubiquitin-specific peptidase 25 (USP25) can modulate inflammation in diseases. This study expounded on the role of USP25 in pyroptosis of acinar cells in AP. Acinar cells were treated with lipopolysaccharide (LPS) and caerulein (CRE) to induce AP. Afterward, the expression patterns of USP25, microRNA (miR)-10a-5p, and Krüppel-like factor 4 (KLF4) in acinar cells were examined. Then, acinar cell viability and levels of NLR family pyrin-domain containing 3 (NLRP3), cleaved caspase-1, cleaved N-terminal gasdermin D (GSDMD-N), interleukin (IL)-1β, and IL-18 were determined. We observed that USP25 was highly expressed in AP models, and silencing USP25 increased cell viability and inhibited pyroptosis of AP acinar cells. The bindings of USP25 to KLF4 and miR-10a-5p to KLF4 and the GSDMD 3′UTR sequence were validated. We found that USP25 binding to KLF4 inhibited ubiquitination degradation of KLF4, KLF4 transcriptionally decreased miR-10a-5p expression, and miR-10a-5p targeted GSDMD expression. Finally, rescue experiments proved that KLF4 overexpression or miR-10a-5p suppression enhanced pyroptosis of AP acinar cells. Overall, USP25 stabilized KLF4 expression through deubiquitination, limited miR-10a-5p expression, and increased GSDMD expression, finally promoting pyroptosis of acinar cells in AP.

急性胰腺炎（AP）是一种消化系统炎症相关疾病。泛素特异性肽酶 25 ( USP25 ) 可以调节疾病中的炎症。本研究阐述了USP25在AP腺泡细胞焦亡中的作用。用脂多糖 (LPS) 和雨伞素 (CRE) 处理腺泡细胞以诱导 AP。随后，检测了腺泡细胞中USP25、微小 RNA (miR)-10a-5p 和 Krüppel 样因子 4 ( KLF4 )的表达模式。然后，测定腺泡细胞活力和 NLR 家族热蛋白结构域 3 (NLRP3)、裂解的 caspase-1、裂解的N末端gasdermin D ( GSDMD - N )、白细胞介素 (IL)-1β 和 IL-18 的水平。我们观察到USP25在 AP 模型中高表达，沉默USP25会增加细胞活力并抑制 AP 腺泡细胞的焦亡。验证了USP25与KLF4以及miR-10a-5p与KLF4和GSDMD 3'UTR 序列的结合。我们发现USP25与KLF4的结合抑制了KLF4的泛素化降解，KLF4转录降低了miR-10a-5p的表达，并且miR-10a-5p靶向GSDMD的表达。最后，拯救实验证明KLF4过表达或miR-10a-5p抑制增强了AP腺泡细胞的焦亡。总体而言，USP25通过去泛素化稳定KLF4表达，限制miR-10a-5p表达，并增加GSDMD表达，最终促进AP中腺泡细胞焦亡。