从JAK3STAT信号途径探讨莪术醇抑制JurkatT细胞增殖的机制.docVIP

从JAK3/STAT信号途径探讨莪术醇抑制Jurkat T细胞增殖的机制 王恒1，王勇1，江晓霁1，王志中2，刘晓飞1，方勇飞1（400038重庆，第三军医大学西南医院中西医结合科1;中国人民解放军第474医院2） [摘要] 目的：观察莪术醇对抑制Jurkat T细胞增殖的影响，并从JAK3-STAT信号途径探讨其作用机制。方法：不同浓度（6.25?g/ml、12.5?g/ml、25?g/ml、50?g/ml）莪术醇干预处理人Jurkat T细胞株，分别用新型四唑单钠盐法、流式细胞技术观察检测细胞增殖活力及细胞周期分布。Hoechst33258染色，观察莪术醇诱导的细胞凋亡形态学变化。设正常对照组、IL-2刺激组、JAK3抑制剂组及莪术醇干预组，Western blot检测细胞中JAK3、STAT3及STAT5a磷酸化蛋白表达量的变化。结果：不同浓度的莪术醇体外处理24h后，Jurkat T细胞的增殖受到明显抑制，呈浓度依赖性下降（P＜0.05）。细胞周期分布结果显示，随着莪术醇作用浓度的增加，细胞凋亡的比例明显升高，S期细胞比例呈浓度依赖性增多，G2/M细胞比例则呈减少趋势。50?g/ml的莪术醇处理24h后，细胞表现出核固缩、碎裂等典型的凋亡形态学变化。Western blot结果显示，50?g/ml的莪术醇可抑制JAK3与STAT5a磷酸化蛋白的表达（P＜0.05），但对STAT3无明显影响（P＞0.05）。结论：莪术醇可能作用于Jurkat T细胞S~G2/M期，阻滞S期细胞向G2/M期移行以抑制细胞增殖。细胞增殖的抑制可能与莪术醇下调JAK3/STAT5信号分子的表达相关。 [关键词] 莪术醇；Jurkat T细胞；JAK3-STAT；细胞增殖 [中图法分类号] R285.5; R593.22; R932 [文献标识码] A [基金项目]重庆市卫生局中医药科研项目（No: 2012-2-62）及国家自然科学基金面上项目（No: [通讯作者]王勇，电话:(023 E-mail: HYPERLINK mailto:wangyongjhy@ wangyongjhy@ Antiproferative effect of curcumol on human T–cell leukemia cell line Jurkat via blocking JAK3/STAT pathway Wang Heng1, Wang Yong1, Jiang Xiaoji1, Wang Zhizhong2, Liu Xiaofei1, Fang Yongfei1 1Department of Integrated Chinese and Western Medicine, the Southwest Hospital of the third Military Medical University, Chongqing 400038, China; 2 No.474 Hospital of Chinese People’s Liberation Army [Abstract] Objective: To observe inhibitory effect of curcumol on Jurkat T cells and explore the potential mechanism from JAK3 (Janus Kinase3)/STAT(signal transducers and activiators of transcription) signal pathway. Methods: Cell viability was assessed by tetrazolium single sodium salt and the characteristics of curcumol-induced cell cycle arrest was detected by flow cytometry (FCM) technologies after exposure to different concentration of curcumol (6.25?g/ml、12.5?g/ml, 25?g/ml and 50?g/ml) for 24h. The cellular morphology of apoptosis induced by curcumol was assessed by Hoechst 33258 staining. The protein expression levels of p-JAK3 (phosphorylated JAK3), p-