MTT法测定棉酚对多浪羊淋巴细胞生长的影响  

The Effect of Gossypol on Lymphocyte Growth of Duolang Sheep by MTT

作　　者：王娟娟 贾山岭[1] 万新军 李莲瑞[2] 

机构地区：[1]塔里木大学生命科学学院,新疆阿拉尔843300 [2]塔里木大学动物科学学院,新疆阿拉尔843300

出　　处：《湖北农业科学》2013年第5期1098-1101,共4页Hubei Agricultural Sciences

基　　金：国家自然科学基金资助项目(30960277)

摘　　要：通过MTT法测定棉酚对多浪羊淋巴细胞生长的影响,确定棉酚对多浪羊淋巴细胞产生影响的最短作用时间和最低浓度,用于指导生产实践。从新疆多浪羊外周血分离淋巴细胞,将其加入10 mL的RPMI-1640中,均匀加入96孔细胞培养板中,于CO2培养箱中37℃培养12 h后,分别以不同浓度(5、10、15、20、25、30、35μmol/L)的棉酚进行作用,作用时间分别为1、2、3、4、5、6 h,加入MTT培养4 h,加入DMSO溶液终止反应,用酶联免疫检测仪于490 nm处检测各孔的吸光值。结果表明,随着棉酚作用浓度及作用时间的递增,多浪羊淋巴细胞活性在逐渐下降,说明棉酚对多浪羊的淋巴细胞活性有明显的抑制作用。The effect of gossypol on the growth of Duolang sheep lymphocyte was detected by MTT assay.The object was to determine the shortest time and lowest concentration of gossypol on lymphocyte,and then used to practice.The lymphocyte was isolated from the peripheral blood of Xinjiang Duolang sheep,then added into 10 mL RPMI-1640 and even allocated to 96-well cell culture plate.After cultured in CO2 incubator at 37 ℃ for 12 h,the lymphocytes were treated with gossypol of different concentrations(5,10,15,20,25,30,35 μmol/L) for 1,2,3,4,5,6 h,and then added MTT to culture 4 h.At last the reaction was stopped by adding DMSO solution.The absorbance of each well was detected by ELISA at OD490 nm.The results showed that the activity of Duolang sheep lymphocytes decreased with the increase of gossypol concentration and acting time.It indicated that gossypol had obvious inhibitory effect on the Xinjiang Duolang sheep lymphocytes.

关 键 词：MTT 棉酚 新疆多浪羊 淋巴细胞 影响 

分 类 号：S[农业科学]