β-HB对牛皱胃平滑肌细胞活性影响——MTT法和CCK-8法的比较研究  

Effect of β-HB on the abomasum smooth muscle cell viability of dairy cow—comparative studies on the MTT and CCK-8 assay

作　　者：谢光洪[1] 许远靖[1] 田武林[1] 李彬[1] 魏腾[1] 张乃生[1] 刘国文[1] 

机构地区：[1]吉林大学动物医学学院,吉林长春130062

出　　处：《中国兽医学报》2013年第3期436-438,449,共4页Chinese Journal of Veterinary Science

基　　金：国家自然科学基金资助项目(31101865)

摘　　要：以牛皱胃平滑肌细胞为研究对象,采用MTT法和CCK-8法检测添加不同浓度β-羟丁酸(β-hydroxybutyricacid,β-HB)对牛皱胃平滑肌细胞活性的影响。通过接种不同的细胞密度((0.2、0.5、1、2、3)×105个/mL),加入MTT/CCK-8孵育后,在不同的波长下读取D值并进行分析。确定CCK-8法的最佳检测波长为450nm,最佳细胞浓度为1×105个/mL,最佳检测时间为4h;MTT法的最佳检测波长为490nm,最佳细胞浓度为3×105个/mL。在检测β-羟丁酸对牛皱胃平滑肌细胞活性的影响时,2种方法的结果对比发现CCK-8法的数据偏差小,准确性高,灵敏,优于MTT法。To investigate the optimal experimental conditions of MTT and CCK-8. Different con- centration abomasum smooth muscle cells（（0.2,0.5,1,2,3） ×10^5/mL） were incubated with dif- ferent concentrations of β-hydroxybutyric acid（β-HB）, and the optimal absorbance wavelenght was measured by CCK-8 or MTT to determine the abomasum smooth muscle cell viability of dairy cow. We found that the optimal absorbance wavelenght was measured at 490 nm and the optimal concentration of cell was 3 ×10^5/mL in the MTT assay,and the optimal absorbance wavelenght was measured at 450 nm,the optimal concentration of cell was 1 ×10^5/mL and the optimal reac- ting time was 4 hours in the CCK-8 assay. These results suggest that CCK-8 assay leads to a smaller deviation,the accuracy and sensitivity is better than MTT assay for determining cell via- bility of abomasum smooth muscle cell pretreated with different concentrations of β-hydroxybu- tyric acid in vitro.

关 键 词：Β-羟丁酸 MTT CCK-8 细胞活力 

分 类 号：S823[农业科学—畜牧学]