莪术醇通过JAK3/STAT信号途径抑制Jurkat细胞增殖  

Proliferation-inhibiting effect of curcumol on human T-cell leukemia cell line Jurkat via blocking JAK3/STAT pathway

作　　者：王恒[1] 王勇[1] 江晓霁[1] 王志中[2] 刘晓飞[1] 方勇飞[1] 

机构地区：[1]第三军医大学西南医院中西医结合科,重庆400038 [2]解放军第474医院康复科,乌鲁木齐83001

出　　处：《第三军医大学学报》2013年第5期408-411,共4页Journal of Third Military Medical University

基　　金：重庆市卫生局中医药科研项目(2012-2-62);国家自然科学基金面上项目(30973827)~~

摘　　要：目的观察莪术醇对抑制Jurkat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、12.5、25、50μg/mL)莪术醇干预处理人Jurkat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。Hoechst 33258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24 h后,Jurkat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。50μg/mL的莪术醇处理24 h后,细胞表现出核固缩、碎裂等典型的凋亡形态学变化。Western blot检测结果显示,50μg/mL的莪术醇可抑制JAK3与STAT5a磷酸化蛋白的表达(P<0.05,P<0.01),但对STAT3无明显影响(P>0.05)。结论莪术醇作用于Jurkat细胞S～G2/M期,阻滞S期细胞向G2/M期移行,并下调JAK3/STAT5信号分子以抑制该细胞增殖。Objective To observe the inhibitory effect of curcumol on Jurkat cells, and to explore the potential mechanism through JAK3 （Janus kinase 3）/STAT （signal transducers and activators 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） after Jurkat cells exposed to different concentrations of curcumol （6.25, 12.5, 25 and 50 μg/mL） for 24 h. The cell morphology of apoptosis induced by curcumol was assessed by Hoechst 33258 staining. The Jurkat cells were divided into four groups including a normal control group, a interleukin-2 （IL-2） stimulation group, a JAK3 inhibitor group and a curcumol intervention group, and the protein expression levels of p-JAK3 （phosphorylated JAK3）, p-STAT3 and p-STAT5a were determined by Western blot analysis. Results After different concentrations of curcumol treatment, the Jurkat cell viability decreased significantly in a dose-dependent manner （P〈0.05）. FCM results showed that apoptotic rate and the proportion of the cells arrested in S phase were elevated along with curcumol concentration increase, while the proportion of cells in G2/M phase declined gradually. After exposure to curcumol for 24 h, the Jurkat cells underwent typical apoptotic morphological changes, such as nucleus condensation and fragmentation. The expression levels of p-JAK3 and p-STAT5a in Jurkat cells were suppressed by 50 μg/mL of curcumol （P〈0.05, P〈0.01）, while there was no obvious change in STAT3 expression （P〉0.05）. Conclusion The underlying mechanism of curcumol on suppressing the proliferation of Jurkat cells may be associated with S to G2/M phase arresting and JAK3/STAT5 pathway down-regulation.

关 键 词：莪术醇 JURKAT T细胞 JAK3-STAT 细胞增殖 

分 类 号：R285.5[医药卫生—中药学] R329.28[医药卫生—中医学]