METHODS: We established four severe acute pancreatitis models of rats including the sham control group, the SAP group, the Fer -1-treated SAP (SAP + Fer-1) group, the 3-MA-treated SAP (SAP + 3-MA) group. The SAP group was induced by retrograde injection of sodium taurocholate into the pancreatic duct. The other two groups were intraperitoneally injected with ferroptosis inhibitor (Fer-1) and autophagy inhibitor (3-MA), respectively. The model of severe acute pancreatitis with amylase crest-related inflammatory factors was successfully established. Then we detected ferroptosis (GPX4, SLC7A1 etc.) and autophagy-related factors (LC3II, p62 ect.) to further clarify the relationship between ferroptosis and autophagy.
RESULTS: Our study found that ferroptosis occurs during the development of SAP, such as iron and lipid peroxidation in pancreatic tissues, decreased levels of reduced glutathione peroxidase 4 (GPX 4) and glutathione (GSH), and increased malondialdehyde(MDA) and significant mitochondrial damage. In addition, ferroptosis related proteins such as GPX4, solute carrier family 7 member 11(SLC7A11) and ferritin heavy chain 1(FTH1) were significantly decreased. Next, the pathogenesis of ferroptosis in SAP was studied. First, treatment with the ferroptosis inhibitor ferrostatin-1(Fer-1) significantly alleviated ferroptosis in SAP. Interestingly, autophagy occurs during the pathogenesis of SAP, and autophagy promotes the occurrence of ferroptosis in SAP. Moreover, 3-methyladenine (3-MA) inhibition of autophagy can significantly reduce iron overload and ferroptosis in SAP.
CONCLUSIONS: Our results suggest that ferroptosis is a novel pathogenesis of SAP and is dependent on autophagy. This study provides a new theoretical basis for the study of SAP.
方法:我们建立了4个大鼠重症急性胰腺炎模型,包括假对照组,SAP集团,Fer-1处理的SAP(SAP+Fer-1)组,3-MA处理的SAP(SAP+3-MA)组。SAP组采用胰管逆行注射牛磺胆酸钠诱导。其他两组均腹腔注射铁凋亡抑制剂(Fer-1)和自噬抑制剂(3-MA),分别。成功建立了淀粉酶相关炎症因子的重症急性胰腺炎模型。然后我们检测到铁凋亡(GPX4,SLC7A1等。)和自噬相关因子(LC3II,p62ect.)进一步阐明铁凋亡与自噬的关系。
结果:我们的研究发现,在SAP的发展过程中会发生铁死亡,例如胰腺组织中的铁和脂质过氧化,还原型谷胱甘肽过氧化物酶4(GPX4)和谷胱甘肽(GSH)的水平降低,并增加丙二醛(MDA)和显著的线粒体损伤。此外,铁凋亡相关蛋白如GPX4、溶质载体家族7成员11(SLC7A11)和铁蛋白重链1(FTH1)均显著降低。接下来,研究了SAP中铁死亡的发病机制。首先,用铁凋亡抑制剂铁抑素-1(Fer-1)治疗可显着减轻SAP的铁凋亡。有趣的是,自噬发生在SAP的发病机制中,自噬促进SAP铁凋亡的发生。此外,3-甲基腺嘌呤(3-MA)抑制自噬可以显着降低SAP中铁过载和铁凋亡。
结论:我们的结果表明,铁凋亡是SAP的一种新的发病机制,并且依赖于自噬。本研究为SAP的研究提供了新的理论基础。