夏枯草对人甲状腺髓样癌细胞增殖的影响和诱导其凋亡的作用机制  

Effects and mechanism of Spica Prunellae on proliferation and apoptosis of human medullary thyroid carcinoma TT cells

作　　者：熊燚[1] 赵敏[2] 谭剑斌[2] 杨哲[3] 张文娟[1] 杨杏芬[2] XIONG Yi;ZHAO Min;TAN Jian-bin;YANG Zhe;ZHANG Wen-juan;YANG Xing-fen(School of Medicine,Jinan University,Guangzhou 510632,China;Guangdong Provincial Center for Disease and Control and Prevention,Guangzhou 511430,China;School of Public Health,Sun Yat-Sen University,Guangzhou 510080,China)

机构地区：[1]暨南大学医学院,广州510632 [2]广东省疾病预防控制中心,广州511430 [3]中山大学公共卫生学院,广州510080

出　　处：《中华中医药杂志》2018年第8期3379-3384,共6页China Journal of Traditional Chinese Medicine and Pharmacy

基　　金：中华人民共和国科学技术部国家高技术研究发展计划(863计划)项目(No.2010AA023001);广东省自然科学基金项目(No.S2013010013289);广东省科技计划项目(No.2013B010404033);广东省中药局课题(No.20132108;No.20121275);广州市科技计划项目(No.201300000161)~~

摘　　要：目的:探索夏枯草对人甲状腺髓样癌(TT)细胞增殖和诱导其凋亡的影响及其可能的作用机制。方法:采用MTT比色法测定2~200mg/mL夏枯草作用24h对TT细胞增殖的影响;采用Hoechst染色法和流式细胞术(Annexin V-FITC/PI联合标记)观察0.425、0.850、1.700、3.400、6.800mg/mL夏枯草作用24h TT细胞凋亡的影响;采用流式细胞术(JC-1荧光染色)观察0.425、0.850、1.700、3.400、6.800mg/mL夏枯草作用24h对TT细胞线粒体膜电位变化的影响;采用蛋白质免疫印迹法(Western blot)测定1.700、3.400mg/mL夏枯草作用24h对TT细胞凋亡相关蛋白表达的影响。结果:在2~200mg/mL的浓度范围内夏枯草作用24h对TT细胞增殖有明显抑制作用(P<0.05),IC50值为3.438mg/mL。流式细胞术检测结果显示夏枯草可诱导TT细胞凋亡,0.425、0.850、1.700、3.400、6.800mg/mL组TT细胞总凋亡率,与对照组相比明显升高(P<0.05);夏枯草可影响TT细胞线粒体膜电位变化,与对照组相比,0.425、1.700、3.400、6.800mg/mL组TT细胞线粒体膜电位显著降低(P<0.05);与对照组相比,1.700、3.400mg/mL夏枯草作用24h胞浆内细胞色素C(Cyto C)表达增多(P<0.01),抑制凋亡蛋白Bcl-2和Caspase-3原蛋白表达减少(P<0.05,P<0.01)。3.400mg/mL组活化的Caspase-3表达显著增多(P<0.01)。结论:夏枯草能抑制TT细胞增殖并诱导其凋亡,其机制可能与活化线粒体凋亡通路有关。Objective： To explore the effect and the possible mechanism of Spica Prunellae on proliferation and apoptosis of the human medullary thyroid carcinoma cells. Methods： The effect on TT cell proliferation of Spica Prunellae in 2~200 mg/ mL was tested by MTT assay after 24 hours. The influence on TT cell apoptosis of different concentrations of Spica Prunellae as 0.425, 0.850, 1.700, 3.400, 6.800mg/mL were observed by method of Hoechst staining and flow cytometry after 24 hours. The influence on mitochondrial membrane potential changes of TT cell of different concentrations of Spica Prunellae as 0.425, 0.850, 1.700, 3.400, 6.800mg/mL were observed by flow cytometry after 24 hours. The effect of different concentrations of Spica Prunellae as 1.700, 3.400mg/mL on expression of apoptosis related protein was detected by Western Blotting after 24 hours. Results： Spica Prunellae could obviously inhibit the proliferation of TT cells in 2-200 mg/mL concentration range （P〈0.05）, and the value of IC50 was 3.438mg/mL. Flow cytometry analysis showed that： 0.425, 0.850, 1.700, 3.400, 6.800mg/mL Spica Prunellae could induce the apoptosis of TT cell, and the apoptotic rates were （8.90±1.01）%, （13.53±1.31）%, （20.87F±0.78）%, （32.37±1.01）% and （42.50±2.97）%, compared with the control group was significantly higher（P〈0.05）. Compared with the control group, Spica Prunellae could lead to changes of mitochondrial membrane potential of TT cell, and the 0.425, 1.700, 3.400, 6.800mg/mL Spica Prunellae could induce the mitochondrial membrane potential of TT cell （P〈0.05）. Compared with control group, Western blotting showed that the expressions of apoptosis protein Bcl-2, Caspase-3 of Spica Prunellae were decreased and the expressions of Cleaved Caspase 3, CytoC were increased. Conclusion： Spica Prnnellae could inhibit the cell proliferation and induce the apoptosis of medullary thyroid carcinoma cells, which might be related to activating mitochondrial apoptosis pathway.

关 键 词：夏枯草 人甲状腺髓样癌细胞 凋亡 线粒体通路 

分 类 号：R285.5[医药卫生—中药学]