芦荟大黄素衍生物AE-YJ通过PI3K-Akt/NF-κB和MAPK/NF-κB途径抑制LPS诱导RAW264.7细胞炎症介质的释放  

AE-YJ,a derivative of Aloe-emodin,inhibits LPS-induced release of inflammatory mediators in RAW264.7 cells via PI3K-Akt/NF-κB and MAPK/NF-κB pathways

作　　者：郭静 尚海[2] 马丽炎 高源[2] 李陵宇 邹忠梅[2] 宛蕾 GUO Jing;SHANG Hai;MA Li-yan;GAO Yuan;LI Ling-yu;ZOU Zhong-mei;WAN Lei(Basic Medical Sciences, Guizhou Medical University, Guizhou 550025, China;Institute of Medicinal Plant Development, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100193, China)

机构地区：[1]贵州医科大学基础医学院,贵阳贵州550025 [2]中国医学科学院北京协和医学院药用植物研究所,北京100193

出　　处：《中国药理学通报》2021年第12期1700-1708,共9页Chinese Pharmacological Bulletin

基　　金：国家重大新药创新科技专项(No 2019ZX09735002);中国医学科学院医学科学创新基金(CIFMS,2016-I2M-3-015)。

摘　　要：目的以RAW264.7细胞为研究对象,采用脂多糖(LPS)诱导RAW264.7细胞建立体外炎症模型,探究芦荟大黄素衍生物AE-YJ的抗炎活性及作用机制。方法MTT法测定RAW264.7细胞活力;Griess法检测RAW264.7细胞的NO释放量;ELISA法检测RAW264.7细胞培养上清液中PGE2、TNF-α、IL-6和IL-1β含量;RT-PCR检查IL-6、IL-1β和COX-2的mRNA表达;Western blot法检测RAW264.7细胞中iNOS、COX-2、IκBα、p65、Akt、ERK、JNK和p38的蛋白表达;免疫荧光法检查p65蛋白的核易位情况。结果相同等剂量下,AE-YJ显示出比AE更强的抗炎活性;AE-YJ减少炎症介质(NO、PGE2、TNF-α、IL-6 IL-1β)的产生,降低iNOS和COX-2的蛋白表达,发挥抗炎活性。AE-YJ减少Akt的磷酸化,降低IκBα降解和p65核易位,阻碍NF-κB的活化;此外,AE-YJ还降低MAPK途径中ERK、JNK、p38的磷酸化。结论AE-YJ的抗炎活性优于AE,其作用机制可能是通过抑制PI3K-Akt/NF-κB和MAPK/NF-κB途径减少LPS诱导的RAW264.7细胞中炎症介质的产生。Aim To use lipopolysaccharide(LPS)to induce RAW264.7 cells to establish an in vitro inflammation model to explore the anti-inflammatory activity and mechanism of the Aloe-emodin derivative AE-YJ,taking RAW264.7 cells as the research object.Methods MTT assay was used to determine the viability of RAW264.7 cells.NO release from RAW264.7 cells was measured by Griess method;ELISA was used to detect PGE2,TNF-α,IL-6 and IL-1βin RAW264.7 cell culture supernatant;RT-PCR was used to check the mRNA expression of IL-6,IL-1βand COX-2;Western blot was used to detect the protein expression of iNOS,COX-2,IκBα,p65,Akt,ERK,JNK and p38 in RAW264.7 cells;Immunofluorescence was employed to check the nuclear translocation of p65 protein.Results Studies showed that AE-YJ showed stronger anti-inflammatory activity than AE did at the same dose.AE-YJ reduced the production of inflammatory mediators(NO,PGE2,TNF-α,IL-6 and IL-1β),exerted anti-inflammatory activity,and down-regulated the expressions of iNOS and COX-2.Mechanism studies showed that AE-YJ reduced the phosphorylation of the Akt signaling pathway,reduced IκBαdegradation and p65 nuclear translocation,and hindered the activation of NF-κB.In addition,AE-YJ also reduced the phosphorylation of ERK,JNK and p38 in MAPK pathway.Conclusion The anti-inflammatory activity of AE-YJ is better than that of AE,and its mechanism may be through inhibition of PI3K-Akt/NF-κB and MAPK/NF-κB pathway to reduce LPS-induced production of inflammatory mediators in RAW264.7 cells.

关 键 词：芦荟大黄素衍生物 RAW264.7 炎症 PI3K-AKT NF-ΚB MAPK 

分 类 号：R-332[医药卫生] R284.1