Department of Pathophysiology, Shaanxi University of Chinese Medicine, Xianyang 712046, Shaanxi Province, China.
Medical Experiment Center, Shaanxi University of Chinese Medicine, Xianyang 712046, Shaanxi Province, China.
World J Gastroenterol. 2017 Oct 28;23(40):7242-7252. doi: 10.3748/wjg.v23.i40.7242.
To explore the role of macrophages in chronic pancreatitis (CP) and the effect of Dachaihu decoction (DCHD) on pancreatic fibrosis in mice.
KunMing mice were randomly divided into a control group, CP group, and DCHD group. In the CP and DCHD groups, mice were intraperitoneally injected with 20% L-arginine (3 g/kg twice 1 d/wk for 6 wk). Mice in the DCHD group were administered DCHD intragastrically at a dose of 14 g/kg/d 1 wk after CP induction. At 2 wk, 4 wk and 6 wk post-modeling, the morphology of the pancreas was observed using hematoxylin and eosin, and Masson staining. Interleukin-6 (IL-6) serum levels were assayed using an enzyme-linked immunosorbent assay. Double immunofluorescence staining was performed to observe the co-expression of F4/80 and IL-6 in the pancreas. Inflammatory factors including monocyte chemoattractant protein-1 (MCP-1), macrophage inflammatory protein-1α (MIP-1α) and IL-6 were determined using real time-polymerase chain reaction. Western blot analysis was used to detect fibronectin levels in the pancreas.
Compared with the control group, mice with 20% L-arginine-induced CP had obvious macrophage infiltration and a higher level of fibrosis. IL-6 serum concentrations were significantly increased. Double immunofluorescence staining showed that IL-6 and F4/80 were co-expressed in the pancreas. With the administration of DCHD, the infiltration of macrophages and degree of fibrosis in the pancreas were significantly attenuated; IL-6, MCP-1 and MIP-1α mRNA, and fibronectin levels were reduced.
The dominant role of macrophages in the development of CP was mainly related to IL-6 production. DCHD was effective in ameliorating pancreatic fibrosis by inhibiting macrophage infiltration and inflammatory factor secretion in the pancreas.
探讨巨噬细胞在慢性胰腺炎(CP)中的作用及大柴胡汤(DCHD)对小鼠胰腺纤维化的影响。
昆明小鼠随机分为对照组、CP 组和 DCHD 组。CP 组和 DCHD 组小鼠腹腔注射 20%L-精氨酸(3 g/kg,每周 2 次,共 6 周)。CP 诱导后 1 周,DCHD 组小鼠给予 DCHD 灌胃,剂量为 14 g/kg/d。建模后 2、4、6 周,采用苏木精-伊红和 Masson 染色观察胰腺形态。采用酶联免疫吸附试验测定血清白细胞介素-6(IL-6)水平。采用双免疫荧光染色观察胰腺中 F4/80 和 IL-6 的共表达。采用实时聚合酶链反应检测单核细胞趋化蛋白-1(MCP-1)、巨噬细胞炎性蛋白-1α(MIP-1α)和 IL-6 等炎症因子。采用 Western blot 分析检测胰腺中纤维连接蛋白水平。
与对照组相比,20%L-精氨酸诱导的 CP 小鼠巨噬细胞浸润明显,纤维化程度更高。IL-6 血清浓度显著升高。双免疫荧光染色显示,IL-6 和 F4/80 在胰腺中共同表达。给予 DCHD 治疗后,胰腺巨噬细胞浸润和纤维化程度明显减轻;IL-6、MCP-1 和 MIP-1αmRNA 及纤维连接蛋白水平降低。
巨噬细胞在 CP 发展中的主导作用主要与 IL-6 的产生有关。DCHD 通过抑制胰腺中巨噬细胞浸润和炎症因子的分泌,有效改善胰腺纤维化。