Sayed Nermein F El, Ragab Diaa, Abdo Walied, Ghoneim Mai El-Sayed
Department of Pharmacology and Toxicology, Faculty of Pharmacy, Ahram Canadian University, Giza, Egypt.
Department of Pharmacology and Toxicology, Faculty of Pharmacy, University of Sadat City (USC), Sadat City, 32897, Egypt.
Naunyn Schmiedebergs Arch Pharmacol. 2025 May 1. doi: 10.1007/s00210-025-04157-0.
Intestinal ischemia/reperfusion (IIR) is a substantial cause of mortality and morbidity worldwide. Octreotide (OCT) has been proven to be effective against various organ insults. However, the exact mechanism by which it exerts protective effect against IIR is still obscure. Thus, the aim was to unveil the potential role of octreotide in an IIR model and decipher its mechanism of action. The rats were allocated into sham-operated, IIR, and OCT groups. Histopathological changes were performed to assess the intestinal injury. Immunohistochemical analysis was used to estimate the NF-κB, Bcl2, caspase-3, IL-17, LC3B, and beclin-1. The mRNA of TNF-α and IL-17 were examined using real time PCR. The levels of p-Nrf2, PRX2, p-JNK, ASK1, and LC3 were assessed using western blot technique. The levels of total antioxidant capacity and SOD were measured using appropriate kits. Furthermore, the protein expressions of Bax, caspase-3, ASK1, and Nrf2 were assessed using proper ELISA kits. Additionally, the comet assay was determined to investigate the effect on apoptosis. At the molecular level, OCT administration upregulated TAC and SOD levels, demonstrating its antioxidant effect. The anti-apoptotic effect was signified by the upregulation of Bcl2 and downregulation of Bax and caspase-3, which was confirmed by comet assay. Furthermore, OCT decreased the levels of TNF-α, NF-κB, and IL-17, confirming its anti-inflammatory effect. OCT pre-treatment triggered autophagy, as evidenced by the upregulation of beclin-1 and LC3B. These effects were accomplished by increasing p-Nrf2 and PRX2 and decreasing ASK1 and p-JNK. Consequently, this impeded the necrosis of intestinal cells and improved the intestinal histoarchitecture abnormalities. Ultimately, OCT successfully ameliorated IIR injury via modulating the Nrf2/PRX2/ASK1/JNK signaling trajectory, leading to autophagic, antioxidant, anti-apoptotic, and anti-inflammatory effects.
肠道缺血/再灌注（IIR）是全球范围内导致死亡和发病的重要原因。奥曲肽（OCT）已被证明对各种器官损伤有效。然而，其对IIR发挥保护作用的确切机制仍不清楚。因此，本研究旨在揭示奥曲肽在IIR模型中的潜在作用，并阐明其作用机制。将大鼠分为假手术组、IIR组和OCT组。通过组织病理学变化评估肠道损伤。采用免疫组织化学分析评估核因子κB（NF-κB）、B细胞淋巴瘤-2（Bcl2）、半胱天冬酶-3（caspase-3）、白细胞介素-17（IL-17）、微管相关蛋白轻链3（LC3B）和自噬相关蛋白1（beclin-1）。使用实时聚合酶链反应检测肿瘤坏死因子-α（TNF-α）和IL-17的信使核糖核酸。采用蛋白质印迹技术评估磷酸化核因子E2相关因子2（p-Nrf2）、过氧化物还原酶2（PRX2）、磷酸化应激活化蛋白激酶（p-JNK）、凋亡信号调节激酶1（ASK1）和LC3的水平。使用适当的试剂盒测量总抗氧化能力和超氧化物歧化酶（SOD）的水平。此外，使用适当的酶联免疫吸附测定试剂盒评估Bax、caspase-3、ASK1和Nrf2的蛋白表达。另外，通过彗星试验确定对细胞凋亡的影响。在分子水平上，给予OCT可上调总抗氧化能力和SOD水平，表明其具有抗氧化作用。Bcl2上调以及Bax和caspase-3下调表明其具有抗凋亡作用，彗星试验证实了这一点。此外，OCT降低了TNF-α、NF-κB和IL-17的水平，证实其具有抗炎作用。OCT预处理引发自噬，beclin-1和LC3B上调证明了这一点。这些作用是通过增加p-Nrf2和PRX2以及降低ASK1和p-JNK来实现的。因此，这抑制了肠道细胞的坏死并改善了肠道组织结构异常。最终，OCT通过调节Nrf2/PRX2/ASK1/JNK信号通路成功减轻了IIR损伤，产生自噬、抗氧化、抗凋亡和抗炎作用。
