【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。更多还原

【Abstract】 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, 6.800 mg/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, 6.800 mg/mL were observed by flow cytometry after 24 hours. The effect of different concentrations of Spica Prunellae as 1.700, 3.400 mg/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 3.438 mg/mL. Flow cytometry analysis showed that: 0.425, 0.850, 1.700, 3.400, 6.800 mg/mL Spica Prunellae could induce the apoptosis of TT cell, and the apoptotic rates were(8.90±1.01)%,(13.53±1.31)%,(20.87±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, 6.800 mg/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, Cyto C were increased. Conclusion: Spica Prunellae could inhibit the cell proliferation and induce the apoptosis of medullary thyroid carcinoma cells, which might be related to activating mitochondrial apoptosis pathway.更多还原