灵芝酸G诱导Vβ3^+CD8^+T细胞抗肿瘤活性的研究  

Study on Antitumor Activity of Vβ3^+CD8^+T Cells Induced by Ganoderma Lucidum G

作　　者：余福恒 胡芳[2] 汪宏良[2] 李仲娟[2] 李超[2] 柯萧韵[2] 陈敏[2] 

机构地区：[1]黄石市东方卫生社区服务中心,湖北黄石435000 [2]湖北理工学院医学院,湖北黄石435003

出　　处：《湖北理工学院学报》2016年第4期54-57,共4页Journal of Hubei Polytechnic University

基　　金：湖北省科技项目(项目编号:2011Cdc116);黄石市科技项目(黄科农〔2012〕1号);教育部46批留学归国启动资金支持(46-1)

摘　　要：为探讨SEA(Staphylococcus Enterotoxin A,SEA)存在下的灵芝酸G(Licochalcone-G,LG)对小鼠CD8+细胞活化及杀伤肿瘤细胞的作用,采用流式细胞技术分别检测CD8+细胞表面分子表达及活化后细胞内的颗粒酶B(Granzyme B,Gr B)和穿孔素(Perforin,Pf)的水平、Caspase-3酶的活性分析肿瘤细胞凋亡。结果为:SEA组和SEA+LG组经过3 d刺激能激活小鼠CD8+细胞Vβ3和Vβ11分别为7.4±0.45,11.5±0.72。体外实验显示,SEA组和SEA+LG组的Vβ3+CD8+T细胞内Gr B%的水平分别为70.2±8.04,91.6±8.16;Pf%的水平分别为67.3±6.86,86.4±7.17,与对照组相比有显著性差异(t=5.24,t=6.43;t=4.31,t=5.47,P<0.01);SEA组和SEA+LG组的HL-60肿瘤细胞内Caspase-3酶的活性分别为37.6±4.81,68.8±6.82,与对照组相比有显著性差异(t=3.18,t=5.27,P<0.01);SEA组和SEA+LG组的A20肿瘤细胞内Caspase-3酶的活性分别为23.7±3.58,54.1±5.71,与对照组相比有显著性差异(t=4.23,t=4.88,P<0.01)。SEA依赖的LG能促使小鼠CD8+T细胞Vβ3和Vβ11分子活化,活化的Vβ3+CD8+T细胞内分泌大量的Gr B和Pf,并通过Caspase-3的信号途径诱导肿瘤细胞凋亡。Objective ： To explore the effect of Staphylococcus Enterotoxin A （SEA） dependent ganoderic acid Licochalcone - G （LG） on CD8 ＋ cell activation and killing tumor cells in mouse. Method ： The levels of intracellular Granzyme B（GrB） and Perforin （Pf）, after surface molecules on CD8 ＋ cells were expressed and activated, detected by flow cytometry. Apoptosis of tumor cells was analyzed by the detection of Caspase - 3 En- zyme activity. Result：After SEA group and SEA ＋ LG group had been stimulated for 3 days,Vβ3 and Vβ11 of CD8 ＋ cells in mouse could be activated, and were 7.4 ± 0.45 and 11.5 ± 0. 72 respectively. There were significant difference compared with the control group （t = 5.24, t = 6.43; t = 4.31, t = 5.47, P 〈 0. 01 ）. GrB activity in HL -60 tumor cells of SEA group and SEA ＋ LG group of were 70.2 ±8.04 and 91.6 ±8.16 respectively, and had significant difference compared with the control group （ t = 5.24, t = 6. 43, P 〈 0.01 ） Pf activity in A20 tumor cells of SEA group and SEA LG group were 67.3 ±6.86 and 86.4 ± 7.17respectively, and had significant difference compared with the control group （t = 4. 31, t = 5.47, P 〈 0. 01 ）. HL -60 Caspase - 3 Enzyme activity in tumor ceils of SEA group and SEA ＋ LG group were 37.6 ±4.81 and 68.8 ± 6.82 respectively, and had significant difference compared with the control group （t = 3. 18, t = 5. 27, P 〈0. 01 ）. Caspase-3 Enzyme activity in A20 tumor cells of SEA group and SEA＋ LG group were 23.7± 3.58 and 54.1 ± 5.71 respectively, and had significant difference compared with the control group （t = 4. 23, t =4. 88,P 〈0. 01 ）. Conclusion：SEA dependent on LG can prompt Vβ3 and Vβ11 of CD8＋ cells in mouse to be activated. Large amount of GrB and Pf were excreted in activated Vβ3 ＋ CD8 ＋ T cells and induced tumor cell apoptosis through Caspase -3 signaling pathway.

关 键 词：颗粒酶B 灵芝酸G 肿瘤细胞 

分 类 号：R967[医药卫生—药理学]