摘要
以东北野猪成纤维细胞为研究对象，利用实时荧光定量PCR技术，分析4、15、25和32℃不同温度冷应激条件下细胞内HSPT0mRNA表达变化规律。结果显示在冷应激的低温处理过程中，HSP70mRNA转录量在各温度均为见显著增加；在冷应激的复温培养过程中，HSP70mRNA转录量在4℃和15℃处理后4h后，复温培养的4～8h显著增加（P〈O．05）；在25℃和32℃处理后4h后，复温培养未能诱导细胞内HSP70mRNA转录量显著增加（P〉0．05）；细胞在4℃和15℃处理2、4、6和8h后，复温培养4h，HSP70mRNA转录量随处理温度的减低和时间的延长而增加，处理4～8h组显著高于对照组（P〈0．05），在25℃和32℃处理2、4、6和8h后，复温培养未能显著诱导HSP70mRNA的转录（P〉0．05）。结果表明，冷应激诱导东北野猪成纤维细胞内HSP70mRNA转录量的增加，不是发生在低温处理的应激阶段，而是发生在复温后的细胞应激阶段，温和冷应激（25～32℃）未能诱导复温后HSP70mRNA转录量的显著增加，强度冷应激（4～15℃）诱导复温后HSP70mRNA转录量的显著增加，且与冷应激的强度和时间成正比。
The dynamic expression of HSP70 in the northeast boar fibroblasts was got utilizing real- time fluorescent quantification reverse transcriptase PCR （FQ-RT-PCR）. Cells were preincubated at 4,15,25 and 32℃ for 2,4,6 and 8 h. The results showed that the level of HSP70 transcription at different temperatures is a significant increase cells were preincubated at 4,15,25 and 32℃ for 4 h followed by recovery and incubation at 38℃ for 4 h,the preincubated cells at 4 and 15℃ gave a singnificantly promoted transcription of HSP70 mRNA compared with the control group （P 0.05） ,25 and 32℃group had a tendency to promoted transcription of HSP70 mRNA compared with the control group,but did not reach significant level （P〈0.05）. Preincubation of cells at 4, 15,25 and 32℃ for 2,4,6 and 8 h followed by recovery and incubation at 38℃ for 4 h showed the relative abundance of HSPT0 was propprtional to the time of preincubation at 4℃ and 15℃,at 25℃ and 32℃ transcription of HSP70 compared with the control group no significant difference（P〉0.05） . These results indicated the cold stress-induced increase of HSP70 expression level in fibroblasts northeast wild boar, did not occur during low temperature stress, but in the cellular stress phase after rewarming. Mild cold stress （25-32℃） can not induce a significant increase in HSP70 expression, harsh cold stress （4-15℃） induction of HSPT0 expression increased significant- ly,the degree of cellular stress response was directly related to intensity and duration of cold expo- sure.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2013年第12期1927-1931,1936,共6页
Chinese Journal of Veterinary Science
基金
转基因生物新品种培育重大专项资助项目(2008ZX08006-003)
国家自然科学基金资助项目(31101700)
