甘丙肽体外抑制脂多糖诱导的小鼠RAW264.7巨噬细胞促炎功能研究

doi:10.3969/j.issn.1672-5069.2016.03.005

实用肝脏病杂志 ›› 2016, Vol. 19 ›› Issue (3): 270-274.doi: 10.3969/j.issn.1672-5069.2016.03.005

甘丙肽体外抑制脂多糖诱导的小鼠RAW264.7巨噬细胞促炎功能研究

何玲楠，李兵航，周达，梁灿灿，丁永年，陈源文，范建高

200092 上海市交通大学医学院附属新华医院消化内科（何玲楠，李兵航，周达，陈源文，范建高）；新疆医科大学第二附属医院消化内科（梁灿灿，丁永年）

收稿日期:2015-11-09 出版日期:2016-05-10 发布日期:2016-05-20
通讯作者: 陈源文，E-mail：shsmus@263.net
作者简介:何玲楠，女，23岁，硕士研究生。主要从事脂肪性肝病与肝纤维化形成机制研究。E-mail：henni6789@163.com
基金资助:
新疆维吾尔自治区高校科研计划重点项目（编号：XJEDU2012I001）；国家自然科学基金项目（编号：81170410/81260081）；上海市卫生系统优秀青年人才培养计划项目（编号：XYQ2011010）

Inhibitory effect of galanin on proinflammatory response induced by lipopolysaccharides in a mouse RAW264.7 macrophage cell line in vitro

He Lingnan，Li Binghang，Zhou Da，et al.

Department of Gastroenterology，Xinhua Hospital，Jiaotong University School of Medicine，Shanghai 200092，China

Received:2015-11-09 Online:2016-05-10 Published:2016-05-20

摘要/Abstract

摘要： 目的探讨甘丙肽对小鼠RAW264.7巨噬细胞功能的影响。方法取小鼠RAW264.7巨噬细胞体外培养，分为对照组、脂多糖（LPS）处理组和LPS联合甘丙肽处理组。给予后两组细胞LPS（1μg/ml）刺激2h，使细胞激活，其中一组再加入甘丙肽1000nmol/L继续处理24h，镜下观察各组细胞形态变化；采用荧光定量PCR法检测培养上清TNF-α、IL-6和IL-1βmRNA水平；采用Western blot法检测细胞诱导型一氧化氮合酶（iNOS）和精氨酸酶1（Arg1）蛋白表达水平。结果 LPS处理组IL-6、IL-1β和TNF-αmRNA分别较对照组升高（7083±432.5）倍、（3106±104.3）倍和（108.0±47.58）倍，而甘丙肽处理组三者水平较LPS处理组下降（1.2±0.2）倍、（0.3±0.08）倍和（19±1.3）倍，差异均有统计学意义（P<0.05）；LPS组iNOS蛋白较对照组升高（19.5±1.964）倍，Arg1蛋白水平较对照组降低（1.6±0.3）倍，而给予甘丙肽混合处理后，iNOS蛋白水平较LPS处理组降低（1.7±0.32）倍，而Arg1蛋白升高（2.31±0.12）倍，差异均有统计学意义（P<0.05）。结论甘丙肽可抑制LPS诱导的小鼠Raw264.7巨噬细胞的促炎反应。

关键词: RAW264.7巨噬细胞系, 甘丙肽, 细胞因子, 精氨酸酶1, 体外

Abstract: Objective To investigate the effect of galanin on mouse RAW264.7 macrophage cell line. Methods RAW264.7 mouse macrophage cells were cultured in DMEM （100 U/ml of penicillin and 100g/ml of streptomycin) containing 10% FBS at 37℃，in a humidified 5% CO2 atmosphere. Cells were divided into three groups，namely the control group （vehicle only)，lipopolysaccharides （LPS） group（LPS only） and galanin treatment group （LPS plus galanin）. Cells were serum-starved for 4 h followed by LPS（1μg/ml） or vehicle incubation for 2 h；Cell were then incubated with or without galanin（1000nmol/L） for 24 h. Cell morphology was observed under the microscope；mRNA levels of tumor necrosis factor-α（TNF-α），interleukin-6（IL-6） and interleukin-1β（IL-1β）were measured by real-time RT-PCR and Western blotting analysis were performed to determine the inducible nitric oxide synthase（iNOS） and arginase 1（Arg1）protein levels. Results The mRNA levels of IL-6，IL-1β and TNF-α in LPS group was increased by（7083 ± 432.5），（3106 ± 104.3） and（108.0±47.58） folds of that in the control group，respectively（P<0.05）；and the mRNA levels of IL-6，IL-1β and TNF-α in cells treated with galanin decreased by（1.2±0.2），（0.3±0.08） and（19±1.3） folds，respectively，as compared with LPS group（P<0.05）；Relative level of iNOS protein in LPS group increased by（19.5±1.964） folds compared with that of the control group，and relative level of Arg1 protein in LPS group was decreased by（1.6±0.3） folds compared with that of the control group；After galanin treatment， iNOS protein level decreased by（1.7±0.32） folds and Arg1 protein increased by （2.31±0.12） folds lower or greater than that treated with LPS alone（P<0.05）. Conclusions Galanin exhibits a significant inhibitory effect on proinflammatory response induced by LPS in mouse Raw264.7macrophage in vitro.

Key words: RAW264.7 macrophage cell line, Galanin, Cytokines, Arginase 1, In vitro

何玲楠，李兵航，周达，梁灿灿，丁永年，陈源文，范建高. 甘丙肽体外抑制脂多糖诱导的小鼠RAW264.7巨噬细胞促炎功能研究[J]. 实用肝脏病杂志, 2016, 19(3): 270-274.

He Lingnan,Li Binghang,Zhou Da,et al.. Inhibitory effect of galanin on proinflammatory response induced by lipopolysaccharides in a mouse RAW264.7 macrophage cell line in vitro[J]. JOURNAL OF PRACTICAL HEPATOLOGY, 2016, 19(3): 270-274.

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甘丙肽体外抑制脂多糖诱导的小鼠RAW264.7巨噬细胞促炎功能研究

Inhibitory effect of galanin on proinflammatory response induced by lipopolysaccharides in a mouse RAW264.7 macrophage cell line in vitro

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