Department of Hepatobiliary and Laparoscopic Surgery, Renmin Hospital, Wuhan University, Hubei Key Laboratory of Digestive System Disease, Wuhan, 430060, Hubei Province, China.
Department of Anesthesiology, Renmin Hospital, Wuhan University, Wuhan, 430060, Hubei Province, China.
Dig Dis Sci. 2019 Mar;64(3):759-772. doi: 10.1007/s10620-018-5379-7. Epub 2018 Nov 22.
Macrophage migration inhibitory factor (MIF) is involved in many acute and chronic inflammatory diseases. However, its role in intrahepatic bile duct (IBD) cell damage associated with severe acute pancreatitis (SAP) remains unclear.
This study was aimed to identify the role of MIF and its underlying mechanisms in SAP complicated by IBD cell damage.
Forty-eight specific-pathogen-free male Wistar rats were randomly divided into four groups (N = 12): a sham operation group (SO group) and three SAP model groups (SAP-3h, SAP-6h, and SAP-12h). Immunohistochemistry was used to detect the expression of MIF and P38 in IBD cells. MIF mRNA expression in IBD cells was observed using real-time fluorescent quantitative polymerase chain reaction (real-time PCR). In addition, Western blotting was performed to detect the protein expression of P38, phosphorylated P38 (P-P38), nuclear factor-κB (NF-κB p65), and tumor necrosis factor alpha (TNF-α). Enzyme-linked immunosorbent assays were used to analyze the levels of TNF-α, IL-1β, and IL-6 in the IBD of rats.
Compared with the SO group, the expression of MIF in the IBD was significantly upregulated both at mRNA and at protein levels in the SAP group. Besides, the protein expression levels of P38, P-P38, NF-κB, p65, TNF-α, IL-1β, and IL-6 in the IBD in rats were also significantly increased in the SAP group and the levels increased gradually as acute pancreatitis progressed (all P < 0.05).
MIF may promote the IBD injury and inflammatory reaction in SAP via activating the P38-MAPK and NF-κB signaling pathways.
巨噬细胞移动抑制因子（MIF）参与多种急、慢性炎症性疾病。然而，其在与重症急性胰腺炎（SAP）相关的肝内胆管（IBD）细胞损伤中的作用尚不清楚。
本研究旨在确定 MIF 及其在 SAP 合并 IBD 细胞损伤中的作用机制。
48 只无特定病原体雄性 Wistar 大鼠随机分为 4 组（n=12）：假手术组（SO 组）和 3 个 SAP 模型组（SAP-3h、SAP-6h 和 SAP-12h）。免疫组化法检测 IBD 细胞中 MIF 和 P38 的表达。实时荧光定量聚合酶链反应（real-time PCR）观察 IBD 细胞中 MIF mRNA 的表达。此外，采用 Western blot 检测 P38、磷酸化 P38（P-P38）、核因子-κB（NF-κB p65）和肿瘤坏死因子-α（TNF-α）的蛋白表达。酶联免疫吸附试验分析大鼠 IBD 中 TNF-α、IL-1β 和 IL-6 的水平。
与 SO 组相比，SAP 组 IBD 中 MIF 的 mRNA 和蛋白表达均显著上调。此外，SAP 组大鼠 IBD 中 P38、P-P38、NF-κB p65、TNF-α、IL-1β 和 IL-6 的蛋白表达水平也显著升高，且随着急性胰腺炎的进展逐渐升高（均 P<0.05）。
MIF 可能通过激活 P38-MAPK 和 NF-κB 信号通路，促进 SAP 中 IBD 损伤和炎症反应。
