6-姜酚对脂多糖致RAW264.7细胞炎症模型的作用及机制研究  

Anti-Inflammatory Effect of 6-Gingerol on LPS-induced RAW264.7 Cells and Its Mechanism

作　　者：谢燕飞[1] 王冉[1] 朱金华[1] 周军[1] Xie Yanfei;Wang Ran;Zhu Jinhua;Zhou Jun(Jiangxi University of Traditional Chinese Medicine,Nanchang 330004)

机构地区：[1]江西中医药大学,南昌330004

出　　处：《中药药理与临床》2021年第3期72-75,共4页Pharmacology and Clinics of Chinese Materia Medica

基　　金：国家自然科学基金地区项目(编号:81860815);江西中医药大学博士科研启动基金项目(编号:2018WBZR005);江西省中医药科技计划项目(编号:2019B030);江西省“双一流”学科(中医学)建设资助项目,中医基础理论专项研究(JXSYLXK-ZHYI054)。

摘　　要：目的:采用脂多糖(LPS)诱导RAW264.7细胞构建体外炎症模型,通过检测炎症相关因子的变化,研究6-姜酚的抗炎作用,并从MAPK信号通路角度探讨其可能的分子机制。方法:采用MTT法确定6-姜酚的安全剂量范围;用LPS刺激RAW264.7细胞建立炎症模型,使用Griess法检测细胞培养液中一氧化氮(NO)的含量变化;酶联免疫吸附(Elisa)法测定细胞因子IL-6的分泌变化;实时荧光定量PCR(RT-qPCR)检测细胞中白介素-6(Il-6)、诱导型一氧化氮合酶(Inos)、环氧合酶2(Cox-2)等炎症因子的转录水平;蛋白免疫印迹(Western Blot)法测定P-Erk/Erk、p-JNK/JNK、P-P38/P38蛋白的表达。结果:6-姜酚浓度在20μmol/L以下对RAW264.7细胞的增殖无明显影响,故选取5、10、15、20μmol/L作为6-姜酚的实验剂量。与空白对照组比较,模型对照组NO释放,IL-6含量、iNOS、COX-2、IL-6的转录水平,以及P-Erk/Erk、p-JNK/JNK、P-P38/P38比值均明显升高(P<0.05);与模型对照组比较,6-姜酚各剂量组的NO释放、IL-6含量降低;Il-6、Inos、Cox-2的转录水平降低,P-Erk/Erk、p-JNK/JNK、P-P38/P38比值明显降低(P<0.05)。结论:6-姜酚具有一定的抗炎作用,其分子机制可能与调控MAPK信号通路有关。Objective:Lipopolysaccharide(LPS)was used to induce an inflammation model in RAW264.7 cells in vitro.The anti-inflammatory effect of 6-gingerol was investigated based on inflammatory cytokines,and its possible molecular mechanism was explored from the MAPK signaling pathway.Methods:MTT assay was used to determine the safe dose range of 6-gingerol.Griess assay was used to detect the changes of nitric oxide(NO)in the cell culture supernatant.The secretion level of interleukin(IL)-6 was determined by enzyme-linked immunosorbent assay(ELISA).Real-time quantitative polymerase chain reaction(RT-qPCR)was used to detect the transcription levels of inflammatory cytokines,such as IL-6,inducible nitric oxide synthase(iNOS),and cyclooxygenase(COX)-2.Western blot was used to determine the protein expression of p-Erk/Erk,p-JNK/JNK,and p-p38/p38.Results:The 6-gingerol with a concentration below 20μmol/L showed no significant effect on the proliferation of RAW264.7 cells,so the concentrations of 6-gingerol were determined as 5,10,15,and 20μmol/L for experiments.Compared with the blank control group,the model group showed increased release of NO,IL-6 content,transcription levels of IL-6,iNOS,and COX-2,and ratios of p-Erk/Erk,p-JNK/JNK,and p-P38/P38(P<0.05).Compared with the model group,6-gingerol groups displayed decreased NO release and IL-6 content,reduced transcription levels of Il-6,iNOS,and COX-2,and declining ratios of p-Erk/Erk,p-JNK/JNK,and p-P38/P38(P<0.05).Conclusion:The 6-gingerol has an anti-inflammatory effect,and its molecular mechanism may be related to the regulation of the MAPK signaling pathway.

关 键 词：6-姜酚 RAW264.7细胞 抗炎作用 丝裂原活化蛋白激酶 信号通路 

分 类 号：R285.5[医药卫生—中药学]