摘要
本研究旨在探讨热应激对奶牛乳腺上皮细胞生长、凋亡及热休克蛋白(Heat shock protein,HSP)mRNA转录的影响。本文以体外高温(42℃)作为热应激模型温度,38℃作为对照温度培养奶牛乳腺上皮细胞,并采用台盼蓝染色绘制生长曲线,细胞流式检测细胞凋亡及RT-qPCR检测HSPs mRNA转录。结果表明,42℃高温处理使得奶牛乳腺上皮细胞生长停滞,细胞数量在培养的第2天已显著少于对照组(38℃)(P<0.05),第3～7天极显著的少于对照组(P<0.01);细胞凋亡率在42℃高温培养测定的第3小时达到最大值;42℃高温使得乳腺上皮细胞热休克蛋白HSP27、70、90和热休克因子HSF-1(Heat shock transcriptional factor)的mRNA转录水平上调,分别为对照组的2.72、7.48、2.69和2.71倍,与对照组相比均差异极显著(P<0.01)。结果提示,高温抑制乳腺上皮细胞正常生长,促使细胞凋亡,并诱导乳腺上皮细胞HSF-1和HSPs mRNA的转录表达。
The experiment was conducted to study the effects of heat stress on bovine mammary epithelial cells in vitro.Bovine mammary epithelial cells were exposed to 42 ℃ as the heat stress model and 38 ℃ as control,then cell growth,apoptosis and heat shock protein expression were detected by trypan blue dying,flow cytometry FCM,RT-qPCR and ELISA.The results showed as follows： cell growth stagnated at 42 ℃;the cell number decreased markedly（P0.05） than control 38 ℃ at the second day,and decreased extremely significantly（P0.01） from the third to the seventh day;the apoptosis rate was the highest at the 3rd hour after heat treatment;the mRNA expression level of heat shock protein 27,70,90（HSP27,HSP70,HSP90） and heat shock factor-1（HSF-1） were 2.72,7.48,2.69 and 2.71 folds as control group,separately,which were all significantly higher than control（P0.01）.In conclusion,heat stress could inhibit cell natural growth,induce apoptosis,and upregulate the mRNA of HSPs.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2010年第5期600-607,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
"十一五"国家科技支撑计划(2006BAD04A14)
关键词
热应激
乳腺上皮细胞
凋亡
热休克蛋白
热休克因子
heat stress
mammary epithelial cells
apoptosis
heat shock protein
heat shock factor
作者简介
作者简介：周振峰（1972-），男，山东泰安人，博士，主要从事反刍动物营养与牛奶质量改良研究，E—mail：zzfl23@126．com；
崔瑞莲（1986-），女，山东人，硕士生，主要从事反刍动物营养学研究，Tel：010-62815833，E-mail：crl0611@163．com；
通讯作者：王加启，E—mail：wang-jia—qi@263．net
