Clinical Pharmacy, Central People's Hospital of Zhanjiang, Zhanjiang, PR China.
Surgical Intensive Care Unit, Central People's Hospital of Zhanjiang, Zhanjiang, PR China.
Immunopharmacol Immunotoxicol. 2022 Oct;44(5):757-765. doi: 10.1080/08923973.2022.2082304. Epub 2022 Jun 6.
Fentanyl is an analgesic used against pancreatitis-related pain, while whether it ameliorates severe acute pancreatitis (SAP) has yet to be checked. This study aims to determine fentanyl-delivered effect on SAP and the mechanism underlying this effect.
Rat SAP models were established, following fentanyl treatment. The serum activity of amylase (AMY), lipase (LIP), and diamine oxidase (DAO) was detected by enzyme-linked immunosorbent assay (ELISA). Histological examination was performed in the pancreatic and intestinal tissues with hematoxylin-eosin staining. After transfection with matrix metalloproteinase (MMP) 9 overexpression plasmids, Caco-2 monolayers were treated with fentanyl and subsequently exposed to lipopolysaccharide (LPS). The transepithelial electrical resistance (TEER) value was determined in rat intestinal mucosa through an Ussing chamber assisted by Analyze & Acquire, and in Caco-2 cell monolayers through a voltohmmeter. Intestinal mucosa and paracellular permeabilities were determined by fluorescein isothiocyanate (FITC)-labeled dextran assay. The expressions of ZO-1, Occludin, MMP9, Fas and Fas ligand (FasL) in rat intestinal mucosa and/or Caco-2 monolayers were analyzed by qRT-PCR or/and western blot.
Fentanyl alleviated SAP-related histological alterations in the pancreas and intestines, reduced the elevated levels of SAP-related AMY, LIP, and DAO, but promoted the levels of ZO-1 and Occludin. In SAP rats and Caco-2 monolayers, SAP-related or LPS-induced TEER value decreases, permeability increases, and increases in the expressions of MMP9, Fas, and FasL were reversed partly by fentanyl. Notably, MMP9 overexpression could reverse the above fentanyl-delivered effects.
Fentanyl alleviates intestinal mucosal barrier damage in rats with SAP by inhibiting the MMP9/FasL/Fas pathway.
芬太尼是一种用于治疗胰腺炎相关疼痛的镇痛药，但其是否能改善重症急性胰腺炎（SAP）尚未得到验证。本研究旨在确定芬太尼对 SAP 的作用及其作用机制。
建立大鼠 SAP 模型，给予芬太尼治疗。通过酶联免疫吸附试验（ELISA）检测血清淀粉酶（AMY）、脂肪酶（LIP）和二胺氧化酶（DAO）的活性。用苏木精-伊红染色法对胰腺和肠道组织进行组织学检查。用基质金属蛋白酶（MMP）9 过表达质粒转染后，用芬太尼处理 Caco-2 单层细胞，然后暴露于脂多糖（LPS）下。通过 Analyze & Acquire 辅助 Ussing 室在大鼠肠黏膜中测定跨上皮电阻（TEER）值，通过电压计在 Caco-2 细胞单层中测定。通过荧光素异硫氰酸酯（FITC）标记的葡聚糖测定肠道黏膜和细胞旁通透性。通过 qRT-PCR 和/或 Western blot 分析大鼠肠黏膜和/或 Caco-2 单层中 ZO-1、Occludin、MMP9、Fas 和 Fas 配体（FasL）的表达。
芬太尼缓解了 SAP 相关的胰腺和肠道组织学改变，降低了 SAP 相关的 AMY、LIP 和 DAO 水平，但促进了 ZO-1 和 Occludin 的水平。在 SAP 大鼠和 Caco-2 单层中，SAP 相关或 LPS 诱导的 TEER 值降低、通透性增加以及 MMP9、Fas 和 FasL 的表达增加部分被芬太尼逆转。值得注意的是，MMP9 过表达可逆转芬太尼的上述作用。
芬太尼通过抑制 MMP9/FasL/Fas 通路缓解 SAP 大鼠肠道黏膜屏障损伤。
