摘要
为了揭示热休克蛋白90基因(Heat shock protein 90,HSP90)在细胞冷应激反应过程中的时空表达特性。以东北野猪成纤维细胞为研究对象,采用实时荧光定量PCR技术分析了4、15、25和32℃不同强度冷应激条件下细胞内HSP90mRNA转录变化规律。结果表明,在低温处理过程中,HSP90mRNA转录量在各温度均未见显著增加(P>0.05);在冷应激的复温培养过程中,HSP90mRNA转录量在4和15℃处理4h后复温培养8h内显著增加(P<0.05),峰值出现在6h,在25和32℃处理4h后复温培养8h内均未见显著增加;细胞在4和15℃处理2、4、6和8h后,复温培养4h,HSP90mRNA转录量随处理温度的降低和时间的延长而增加,在25和32℃处理2、4、6和8h后,复温培养未能显著诱导HSP90mRNA的转录。结果提示,冷应激诱导东北野猪成纤维细胞内HSP90mRNA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HSP90mRNA转录量的显著增加,强度冷应激(4～15℃)诱导复温后HSP90mR-NA转录量的显著增加,且与冷应激的强度和时间成正比。
To investigate the spatial and temporal expression profiles of HSP90 gene in fibroblasts of the Northeast Wild boar under cold stress. Real-time fluorescent quantification reverse tran- scriptase PCR （FQ-RT-PCR） was applied to analyze the expression of HsPgo mRNA in the Northeast Wild boar fibroblasts under 4,15,25 and 32 ℃. The results showed that HsPgo mRNA transcrip- tion level didn＇t significantly increase （P〈0.05） under cold stress treatments at different tem- peratures~ During rewarming culture, HsPgo mRNA transcription levels increased significantly （P〈0.05） within 8 h of rewarming incubation following preincubation at 4 or 15 ℃ and the peak showed at the 6~h hour. While HSP90 mRNA transcription levels didn＇t increase significantly in the 25 or 32 ℃ preincubation groups within 8 h of rewarming incubationg HsPgo mRNA tran- scription levels increased with decreasing of the temperature and duration of the cold treatment time when cells were preincubated at 4 or 15 ℃ for 2, 4, 6 or 8 h followed by a 4 h rewarming incubation. However, rewarming incubation didn＇t significantly induce transcription of HSPgO mRNA in cells preincubated at 25 or 32 ℃ for 2, 4, 6 or 8 h. The result indicated that cold stress induced increase of HSPgO mRNA transcription level in fibroblasts of the Northeast Wild boar, did not occur in the stress period during low temperature treatment, but in the cellular stress period after rewarming. Mild cold stress （2S-32 ℃） didn＇t induce a significant increase at HSP90 mRNA transcript levels, while harsh cold stress （4-15 ℃） induced a significant increase at HSP90 mRNA transcript levels which were directly proportional to the intensity and duration of cold exposure at 4-15 ℃.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2012年第12期1978-1983,共6页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
转基因生物新品种培育重大专项(2008ZX08006-003)
国家自然科学基金(31101700)
