秋水仙碱对大鼠急性胰腺炎的作用及其机制研究

Effects of colchicine on acute pancreatitis in rat models and the underlying mechanisms

摘要:

目的 观察秋水仙碱对急性胰腺炎（AP）的作用，并探讨相关机制。方法 将30只AP大鼠随机分为模型组、低剂量组和高剂量组，每组10只。另取10只大鼠仅开腹，翻动胰腺，关腹，不做其他处理，设为对照组。低、高剂量组分别舌下静脉注射秋水仙碱2、4 mg/kg，对照组、模型组均舌下静脉注射等体积0.1%二甲基亚砜溶液，1次/d，连续5 d。采用酶联免疫吸附试验检测血清炎症因子水平；苏木精-伊红染色观察胰腺组织病理变化；Western blotting检测胰腺组织肿瘤坏死因子受体-1（TNF-R1）、TNFR1相关的死亡区域蛋白（TRADD）、受体相互作用蛋白激酶1（RIP1）蛋白的表达。结果 与对照组比较，模型组、低剂量组、高剂量组血清白细胞介素-6（IL-6）、肿瘤坏死因子-α（TNF-α）、丙二醛（MDA）、降钙素基因相关肽（CGRP）水平、病理评分及TNF-R1、TRADD、RIP1蛋白相对表达量升高（P <0.05）；与模型组比较，低剂量组、高剂量组血清IL-6、TNF-α、MDA、CGRP水平、病理评分及TNF-R1、TRADD、RIP1蛋白相对表达量降低（P <0.05）；与低剂量组比较，高剂量组血清IL-6、TNF-α、MDA、CGRP水平、病理评分及TNF-R1、TRADD、RIP1蛋白相对表达量降低（P <0.05）。与对照组比较，模型组、低剂量组、高剂量组血清SOD活性降低（P <0.05）；与模型组比较，低剂量组、高剂量组血清SOD活性增强（P <0.05）；与低剂量组比较，高剂量组血清SOD活性增强（P <0.05）。结论 秋水仙碱可减轻AP大鼠炎症反应、氧化应激，改善微循环，推测其作用机制与抑制TNF-α/TNF-R1信号通路有关。

Abstract:

Objective To observe the effect of colchicine on acute pancreatitis (AP) and to explore the underlying mechanisms.Methods Thirty AP rat models were randomly divided into model group, experimental-L group, and experimental-H group, with 10 rats in each group. Another 10 rats were sham-operated and set as the control group. The rats in the experimental-L group and the experimental-H group were injected sublingually with colchicine at doses of 2 and 4 mg/kg, respectively, while those in the control group and the model group were injected with the same amount of 0.1% dimethyl sulfoxide solution sublingually once a day for 5 consecutive days. The serum levels of inflammatory factors were detected by enzyme-linked immunosorbent assays. The histopathological changes in pancreatic tissues were detected by Hematoxylin-eosin staining. The relative protein expressions of tumor necrosis factor receptor-1 (TNF-R1), TNF-R1-associated death domain protein (TRADD), and receptor interacting protein kinase 1 (RIP1) in pancreatic tissues were detected by Western blotting.Results Compared with the control group, the serum levels of interleukin-6 (IL-6), tumor necrosis factor-α (TNF-α), malondialdehyde (MDA), and calcitonin gene-related peptide (CGRP), the histopathological scores, and the protein expressions of TNF-R1, TRADD and RIP1 were higher in the other groups (P < 0.05). These indicators were lower in the experimental-L group and the experimental-H group than those in the model group (P < 0.05), and they were even lower in the experimental-H group than those in the experimental-L group (P < 0.05). Compared with the control group, the serum SOD activity was lower in the other groups (P < 0.05). In contrast, the serum SOD activity was higher in the experimental-H group than in the experimental-L group (P < 0.05).Conclusions Colchicine can ameliorate the inflammation and oxidative stress and improve the microcirculation in AP rats. It is speculated that the underlying mechanism is dependent on the TNF-α/TNF-R1 signaling pathway.