柴胡皂苷-d对大鼠肝癌细胞H4-ⅡE细胞内Ca^(2+)的影响研究  

Effect of Saikosaponin-d on Intracellular Ca^(2+) Concentration of H4-ⅡE Rat Liver Cells

作　　者：秦勇[1] 陈念平[2] 叶冠雄[1] 徐胜前[1] 吴成军[1] 王世[1] 潘德标[1] 

机构地区：[1]浙江省丽水市人民医院普通外科,323000 [2]广东医学院附属医院肝胆外科,湛江524001

出　　处：《医学研究杂志》2012年第8期103-106,共4页Journal of Medical Research

摘　　要：目的探讨柴胡皂苷-d(saikosaponin-D,SS-D)诱导大鼠肝癌细胞H4-ⅡE细胞内钙离子浓度([Ca2+]cyt)变化的影响研究。方法通过噻唑蓝(MTT)比色实验选择合适的药物浓度、荧光指示剂Fluo-3/AM检测[Ca2+]cyt及钙库中Ca2+的变化。结果 MTT比色结果显示,10、30、50、70、100μmol/L SS-D对细胞的存活率都在90%以上。第一大组中阴性对照组可引起细胞内钙离子浓度升高,其平均荧光强度(MFI)为35.76±1.37;阳性对照组可使其大幅度升高,其MFI值为74.74±1.26。不同浓度的SS-D也可使其不同程度升高,并且与SS-D呈浓度依赖性,其平均荧光强度(MFI)分别为59.95±1.02、73.89±1.35、80.30±0.04、93.30±0.83。阳性对照组及处理组的MFI值明显高于阴性对照组(P<0.05);处理组1、处理组3及处理组4 MFI值与阳性对照组比较差异有统计学意义(P<0.05);石胆酸(LCA)及SS-D、熊脱氧胆酸(UDCA)都能诱导细胞钙库中Ca2+释放。结论 SS-D能诱导细胞内Ca2+内流,增加[Ca2+]cyt,调控细胞内Ca2+的生理活动。Abstract Objective To investigate the influences of saikosaponin - d （SS - D） on the free cytoplasmic concentration of Ca^2＋ in H4 - IIE cells of rat models. Methods Proper concentration of chemicals used in the study was assessed by thiazol blue （MTT） method. Changes in free cytoplasmic Ca^2＋ concentration （ [ Ca^2＋ ] cyt） and intracellular stores Ca^2＋ were measured by Fluo - 3/AM load tech- nique. Results MTT test showed that cell survival rates were all over 90% after treatment of SS - D at the concentration of 10, 30, 50, 70, 100 and 120μmol/L, respectively. The negative control group showed a increase of I Ca^2＋ ] cyt with mean fluorescence intensity （MFI） of 35.76 ± 1.37, and the increase was even more obvious in the positive control group, which showed MFI of 74.74 ± 1.26. For other groups, the MFI of [ Ca^2＋ ] cyt was 59.95 - 1.02,73.89 ± 1.35,80.30 ± 0.04,93.30 -± 0.83, respectively, showing an SS - D dose - dependent increase. MFI were significantly higher in positive control group and the treatment group than that in the negative control group （P 〈 0.05 ）. There was also significant differences in the MFI between treatment group 1 , 3, 4 and that in the positive control group （ P 〈 0. 05）. SS - D, LCA and UDCA can induce the release of Ca^2＋ from intracellular stores. Conclusion SS - D may regulate physio- logical activities of intracellular Ca^2＋ by increasing [ Ca^2＋ ] cyt level since it can induce Ca^2＋ influx.

关 键 词：柴胡皂苷-d钙离子 H4-ⅡE细胞 细胞内钙离子浓度 

分 类 号：R373[医药卫生—病原生物学]