基于TLR4/MyD88/NF-κB信号通路研究黄连素对小鼠巨噬细胞极化的干预作用
x

请在关注微信后,向客服人员索取文件

篇名: 基于TLR4/MyD88/NF-κB信号通路研究黄连素对小鼠巨噬细胞极化的干预作用
TITLE: Intervention Effects of Berberine on Mice Macrophage Polarization Based on TLR 4/MyD88/NF-κB Signaling Pathway
摘要: 目的:基于Toll样受体4(TLR4)/髓样分化因子88(MyD88)/核因子κB(NF-κB)信号通路研究黄连素对小鼠巨噬细胞极化的影响。方法:以小鼠巨噬细胞RAW264.7为对象,以阿托伐他汀钙为阳性对照,经脂多糖(LPS)诱导以复制炎症细胞模型,采用酶联免疫吸附测定法检测低、中、高剂量黄连素(5、10、20μmol/L)作用24h后细胞培养液中肿瘤坏死因子α(TNF-α)、白细胞介素6(IL-6)、NF-κB含量,采用实时荧光定量聚合酶链反应法检测细胞中TLR4、MyD88mRNA的表达水平,采用Westernblotting法检测细胞中TLR4、MyD88、诱导型一氧化氮合酶(iNOS)、CD206蛋白的表达水平。结果:与空白对照组比较,LPS诱导组细胞培养液中TNF-α、IL-6、NF-κB含量,细胞中TLR4、MyD88mRNA的相对表达量以及TLR4、MyD88、iNOS蛋白相对表达量均显著升高(P<0.05)。与LPS诱导组比较,阿托伐他汀钙组和黄连素中、高剂量组TNF-α、IL-6含量,TRL4、MyD88mRNA及其蛋白的相对表达量以及各给药组NF-κB含量和iNOS蛋白的相对表达量均显著降低,且黄连素高剂量组NF-κB含量显著低于阿托伐他汀钙组(P<0.05);阿托伐他汀钙组和黄连素高剂量组CD206蛋白的相对表达量均显著升高,且黄连素高剂量组CD206蛋白的相对表达量显著高于阿托伐他汀钙组(P<0.05)。结论:不同剂量的黄连素均可不同程度地干预小鼠巨噬细胞极化,其机制可能与调控TLR4/MyD88/NF-κB信号通路有关。
ABSTRACT: OBJECTIVE:To study the effects of berberine on mic e macrophage polarization based on TLR 4-MyD88-NF-κB signaling pathway. METHODS :Using mice RAW 264.7 macrophage as the object ,atorvastatin calcium as positive control , inflammatory cell model was induced by lipopolysaccharide (LPS);ELISA method was used to detect the contents of TNF-α,IL-6 and NF-κB in cell culture medium after treated with low,medium and high doses of berberine (5,10,20 μmol/L)for 24 h. The real-time fluorescence quantitative PCR was conducted to determine the mRNA expression of TLR 4 and MyD 88 in cells. Western blotting assay was used to detect the protein expression of TLR 4,MyD88,iNOS and CD 206 in cells. RESULTS :Compared with blank control group ,the contents of TNF-α,IL-6 and NF-κB in cell culture medium,mRNA expression of TLR 4 and MyD 88, protein expression of TLR 4,MyD88 and iNOS in cells were increased significantly in LPS induction group (P<0.05). Compared with LPS induction group ,the contents of TNF-α and IL-6,mRNA and protein expression of TLR 4 and MyD 88 in atorvastatin calcium group ,berberine medium-dose and high-dose groupsas well as the content of NF-κ B and protein expression of iNOS in administration groups were decreased significantly , while the content of NF-κB in berberine high-dose group was significantly lower than atorvastatin calcium group (P<0.05). The protein expressions of CD206 in atorvastatin calcium group and berberine high-dose group were increased significantly ,while the protein expression of CD 206 in berberine high-dose group was significantly higher than atorvastatin calcium group (P<0.05). CONCLUSIONS :Different doses of berberine can intervene in mice macrophage polarization to different extents ,the mechanism of which may be associated with the regulation of TLR4/MyD88/NF-κB signaling pathway.
期刊: 2020年第31卷第15期
作者: 李建功,孙文熙,刘家玥,李雪山,薛伟琪,罗川晋
AUTHORS: LI Jiangong ,SUN Wenxi ,LIU Jiayue,LI Xueshan ,XUE Weiqi,LUO Chuanjin
关键字: 黄连素;小鼠巨噬细胞;RAW264.7细胞;极化;Toll样受体4/髓样分化因子88/核因子κB信号通路
KEYWORDS: Berberine;Mice macrophage ;RAW264.7 cell;Polarization;TLR4/MyD88/NF-κB signaling pathway
阅读数: 77 次
本月下载数: 3 次

* 注:未经本站明确许可,任何网站不得非法盗链资源下载连接及抄袭本站原创内容资源!在此感谢您的支持与合作!