Chen X M, Li F Q, Yan S, Wu X C, Tang C L
Department of Liver Disease, The Second Affiliated Hospital, Zhejiang Chinese Medical University, Hangzhou 310005, China.
Department of Endoscopic Surgery, The First Affiliated Hospital of Whenzhou Medical University, Wenzhou 325000, Zhejiang, China.
Beijing Da Xue Xue Bao Yi Xue Ban. 2016 Oct 18;48(5):777-782.
To investigate the anti-inflammation effects by activation of the cholinergic anti-inflammatory pathway and its mechanisms in non-alcoholic steatohepatitis (NASH) model mice.
6-week-old male C57BL/6J (B6) mice were randomly divided into four groups: the first group was normal mice, injected with saline; the second group was normal mice, injected with nicotine; the third group was NASH model mice, injected with saline; the fourth group was NASH model mice, injected with nicotine. The experimental mice were fed with either standard chow (SC) or high-fat and high-fructose (HFHF) for 17 weeks to generate an NASH model mice. The mice received injection once daily for 3 weeks [nicotine dose, 400 μg/kg]. Then, their pathological characteristics and function of the liver were assessed. The expressions of interleukin-6 (IL-6) and tumor necrosis factor-α (TNF-α) in serum were analyzed by enzyme linked immunosorbent assay (ELISA). The expressions of alpha 7 nicotinic acetylcholine receptors (α7nAChR), Toll-like receptors-4 (TLR-4) and nuclear factor κB of phosphory-lation (p-NF-κB) in Kupffer cells were determined by Western blot and immunofluorescence assays.
We successfully generated NASH model mice by imitating the high-fat and high-fructose dietary style of NASH patients. The results of our investigation demonstrated that nicotine could reduce significantly the levels of IL-6, and TNF-α in serum (P<0.05). The expression of p-NF-κB protein in the group which was NASH model mice injected with nicotine declined significantly as compared with the group which was NASH model mice injected with saline (P<0.05). And the expression of α7nAChR protein elevated significantly conversely (P<0.05).
Activation of the cholinergic anti-inflammatory pathway could inhibit the release of inflammatory factors as TNF-α and IL-6 in NASH model mice, and the mechanism for the inhibition of inflammatory was mediated by NF-κB pathway.
探讨激活胆碱能抗炎通路对非酒精性脂肪性肝炎（NASH）模型小鼠的抗炎作用及其机制。
将6周龄雄性C57BL/6J（B6）小鼠随机分为四组：第一组为正常小鼠，注射生理盐水；第二组为正常小鼠，注射尼古丁；第三组为NASH模型小鼠，注射生理盐水；第四组为NASH模型小鼠，注射尼古丁。对实验小鼠分别给予标准饲料（SC）或高脂高果糖（HFHF）饲料喂养17周以建立NASH模型小鼠。小鼠连续3周每天注射一次[尼古丁剂量，400μg/kg]。然后，评估其病理特征和肝脏功能。采用酶联免疫吸附测定（ELISA）分析血清中白细胞介素-6（IL-6）和肿瘤坏死因子-α（TNF-α）的表达。通过蛋白质免疫印迹法和免疫荧光测定法测定库普弗细胞中α7烟碱型乙酰胆碱受体（α7nAChR）、Toll样受体4（TLR-4）和磷酸化核因子κB（p-NF-κB）的表达。
通过模拟NASH患者的高脂高果糖饮食方式，我们成功建立了NASH模型小鼠。研究结果表明，尼古丁可显著降低血清中IL-6和TNF-α的水平（P<0.05）。与注射生理盐水的NASH模型小鼠组相比，注射尼古丁的NASH模型小鼠组中p-NF-κB蛋白的表达显著下降（P<0.05）。相反，α7nAChR蛋白的表达显著升高（P<0.05）。
激活胆碱能抗炎通路可抑制NASH模型小鼠中TNF-α和IL-6等炎症因子的释放，其抗炎机制是由NF-κB通路介导的。
