Liu Muyun, Ma Lizhe, An Wei, Yang Yaying, Liu Juncen, Jiang Hui, Yuan Jihang, Sun Xiaoru, Zhu Jingyi, Yan Maoyun, Wang Luowei, Li Zhaoshen, Liao Zhuan, Sun Chang
Department of Gastroenterology, First Affiliated Hospital of Naval Medical University, Shanghai, 200433, China; Department of Gastroenterology, NO. 905 Hospital of PLA Navy affiliated to Naval Medical University, Shanghai, 200050, China; Shanghai Institute of Pancreatic Diseases, Shanghai, 200433, China.
Department of Gastroenterology, First Affiliated Hospital of Naval Medical University, Shanghai, 200433, China; Shanghai Institute of Pancreatic Diseases, Shanghai, 200433, China; Department of Gastroenterology, No 988 Hospital of PLA Joint Logistics Support Force, Zhengzhou, 450000, China.
Pancreatology. 2024 Aug;24(5):677-689. doi: 10.1016/j.pan.2024.05.514. Epub 2024 May 7.
BACKGROUND & AIMS: Mutations in genes, including serine protease inhibitor Kazal-type 1 (SPINK1), influence disease progression following sentinel acute pancreatitis event (SAPE) attacks. SPINK1 c.194+2T > C intron mutation is one of the main mutants of SPINK1,which leads to the impairment of SPINK1 function by causing skipping of exon 3. Research on the pathogenesis of SAPE attacks would contribute to the understanding of the outcomes of acute pancreatitis. Therefore, the aim of the study was to clarify the role of SPINK1 c.194+2T > C mutation in the CP progression after an AP attack.
SAPE attacks were induced in wildtype and SPINK mutant (Spink1 c.194+2T > C) mice by cerulein injection. The mice were sacrificed at 24 h, 14 d, 28 d, and 42 d post-SAPE. Data-independent acquisition (DIA) proteomic analysis was performed for the identification of differentially expressed protein in the pancreatic tissues. Functional analyses were performed using THP-1 and HPSCs.
Following SAPE attack, the Spink1 c.194+2T > C mutant mice exhibited a more severe acute pancreatitis phenotype within 24 h. In the chronic phase, the chronic pancreatitis phenotype was more severe in the Spink1 c.194+2T > C mutant mice after SAPE. Proteomic analysis revealed elevated IL-33 level in Spink1 c.194+2T > C mutant mice. Further in vitro analyses revealed that IL-33 induced M2 polarization of macrophages and activation of pancreatic stellate cells.
Spink1 c.194+2T > C mutation plays an important role in the prognosis of patients following SAPE. Heterozygous Spink1 c.194+2T > C mutation promotes the development of chronic pancreatitis after an acute attack in mice through elevated IL-33 level and the induction of M2 polarization in coordination with pancreatic stellate cell activation.
包括丝氨酸蛋白酶抑制剂Kazal型1(SPINK1)在内的基因突变会影响哨兵急性胰腺炎事件(SAPE)发作后的疾病进展。SPINK1 c.194+2T>C内含子突变是SPINK1的主要突变体之一,它通过导致外显子3跳跃而损害SPINK1功能。对SAPE发作的发病机制进行研究将有助于理解急性胰腺炎的结局。因此,本研究的目的是阐明SPINK1 c.194+2T>C突变在急性胰腺炎(AP)发作后慢性胰腺炎(CP)进展中的作用。
通过注射雨蛙素在野生型和SPINK突变体(Spink1 c.194+2T>C)小鼠中诱导SAPE发作。在SAPE发作后的24小时、14天、28天和42天处死小鼠。进行数据非依赖采集(DIA)蛋白质组学分析以鉴定胰腺组织中差异表达的蛋白质。使用THP-1和肝星状细胞(HPSC)进行功能分析。
在SAPE发作后,Spink1 c.194+2T>C突变体小鼠在24小时内表现出更严重的急性胰腺炎表型。在慢性期,SAPE发作后Spink1 c.194+2T>C突变体小鼠的慢性胰腺炎表型更严重。蛋白质组学分析显示Spink1 c.194+2T>C突变体小鼠中白细胞介素-33(IL-33)水平升高。进一步的体外分析表明,IL-33诱导巨噬细胞的M2极化和胰腺星状细胞的激活。
Spink1 c.194+2T>C突变在SAPE发作后患者的预后中起重要作用。杂合子Spink1 c.194+2T>C突变通过升高IL-33水平并协同胰腺星状细胞激活诱导M2极化,促进小鼠急性发作后慢性胰腺炎的发展。