齐墩果酸抑制脂多糖诱导RAW264.7细胞炎症反应  

Inhibition of oleanolic acid on lipopolysaccharide induced RAW264.7 cell inflammatory response

作　　者：吴青青 王娟[1] 崔树娜[1,2] 陈姗姗 雷雨 李士华 钱静 

机构地区：[1]扬州大学医学院,江苏扬州225001 [2]扬州大学附属医院,江苏扬州225001

出　　处：《扬州大学学报（农业与生命科学版）》2017年第3期27-31,共5页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：国家自然科学基金资助项目(81703969);江苏省自然科学基金资助项目(BK20160480)

摘　　要：通过CCK-8法检测齐墩果酸对巨噬细胞增殖的影响;采用Griess试剂检测细胞培养液中NO含量,通过Western Blot法分析iNOS蛋白表达和MAPK通路p38,ERK和JNK蛋白表达和磷酸化状态,研究齐墩果酸对脂多糖(LPS)诱导巨噬细胞RAW264.7炎症反应的影响,探讨齐墩果酸抗感染作用及机制。结果表明:齐墩果酸(100.0μmol·L^(-1))明显抑制巨噬细胞增殖(P<0.05);12.5~100.0μmol·L^(-1)的齐墩果酸能够显著抑制脂多糖诱导巨噬细胞分泌NO(P<0.01),并能显著抑制LPS诱导iNOS蛋白的表达;LPS作用后能够激活ERK通路,抑制p38和JNK通路磷酸化,齐墩果酸作用后,能显著抑制ERK磷酸化,促进JNK和p38蛋白的磷酸化。证明齐墩果酸能显著降低脂多糖诱导巨噬细胞分泌NO,通过抑制iNOS的表达和双向调控MAPK通路蛋白磷酸化达到抗感染作用。CCK-8 assay was used to detect the influence of oleanolic acid （OA） on macrophage proliferation; Griess assay was used to determine the NO release in LPS induced macrophage with or without OA treatment; Western-blot method was used to detect the iNOS protein expression and MAPK pathway p38, ERK and iNK protein expression and phosphorylation status. To study the anti-inflammatory effect of OA on lipopolysaccharide （LPS） induced RAW264.7 macrophage and its mechanism. Results.. only OA （100.0 μmol · L^-1 ） inhibited macrophage proliferation. （P〈0.05） 12. 5--100.0 μmol · L^-1 of OA significantly inhibited the secretion of NO in LPS stimulated macrophages （P〈0. 01）, and strongly inhibited the expression of iNOS protein; LPS activated the phosphorylation of ERK pathway, and inhibited the phosphorylation status of JNK and p38, but OA decreased the phosphorylation of ERK, and promoted the phospho- rylation status of JNK and p38 protein. This study demonstrated that OA showed strong anti-inflammatory effect by inhi- bition of the expression of iNOS and regulation of protein phosphorylation of MAPK pathway in dual way.

关 键 词：齐墩果酸 脂多糖 巨噬细胞RAW264.7 一氧化氮 MAPK通路 

分 类 号：R285.5[医药卫生—中药学]