钙／钙调素依赖性蛋白激酶Ⅱ对热休克小鼠胚胎成纤维细胞HSP70表达的影响

doi:10.3969/j.issn.1000-7091.2012.04.041

摘要/Abstract

摘要： 为证实热休克小鼠胎儿成纤维细胞中钙／钙调素依赖性蛋白激酶Ⅱ（Ca^2＋／calmodulin-dependent protein ki-nase Ⅱ，CaMK Ⅱ）对热休克蛋白70（Heat shock protein70，HSP70）基因表达的影响及其作用机理，将体外培养的小鼠胎儿成纤维细胞（Mouse Embryonic Fibroblasts，MEF）随机分为39℃和41℃热处理组（分别处理0．5，1，1．5．2h）和常温对照组（37℃），测定各组细胞CaMK Ⅱ和HSF1 mRNA的量。此外，将培养的细胞分为37℃和39℃ CaMK Ⅱ特异性抑制剂（myr-AIP）处理组和相应的空白对照组，分别测定各组细胞HSP70、HSF1及其mRNA的量。结果表明，39℃热休克1h后HSF1和CaMK Ⅱ的mRNA表达量极显著高于常温对照组，经过myr—AIP处理的39℃组细胞HSP70及其mRNA含量显著低于39℃空白对照组，而37℃ myr—AIP处理组与37℃对照组没有显著差异；myr-AIP对37℃组和39℃组的HSF1 mRNA和蛋白含量影响均不显著，却使p-HSF1的含量显著降低。热休克能增加CaMK Ⅱ和HSF1 mRNA的转录，且CaMK Ⅱ通过磷酸化HSF1促进HSP70基因表达。

关键词: 热休克反应, HSP70 CaMK Ⅱ HSF1, 小鼠胎儿成纤维细胞

Abstract: The objectives of the study were to verify if the CaMK Ⅱ （Ca^2＋/calmodulin-dependent protein kinase Ⅱ ） influences the expression of HSP70 gene in the mouse embryonic fibroblasts （MEF） with heat shock and to clarify its mechanism. The fibroblasts were heat-treated at 37℃,39℃ and 41℃ individually before the expressions of both CaMK Ⅱ and HSF1 were detected by RT-PCR at 0.5,1,1.5 and 2 h of heat shock, respectivelly. Moreover, the fibroblasts were divided into control group and myr-AIP group randomly, and cultured at 37 ℃ and 39℃,respec- tively. The expressions of both HSP70 and HSF1 were detected. The expressions of HSF1 and CaMK Ⅱ mRNA in the MEF treated at 39℃ for 1 h were both extremely significantly higher （P 〈0. 01 ） than that of the control group. There was a significantly （P 〈 0.05） decrease of HSP70 expression in the MEF cultured at 39℃ treated with myr- AlP compared with its blank control. However,there was no difference in HSP70 exoression between the MEF cul- tured at 37℃ with or without myr-AIP. The expressions of HSF1 in the MEF treated with myr-AIP were not signifi- cantly influenced. However,the p-HSF1 expression in the MEF treated with myr-AIP was significantly （P 〈 0. 05） decreased. Moderate heat shock increases both expressions of CaMK Ⅱ and HSF1 in mouse embryonic fibroblasts. CaMK Ⅱ participates in both heat shock response and HSP70 gene expression by p-HSF1.

Key words: Heat shock response, HSP70 CaMK Ⅱ HSF1, Mouse embryonic fibroblasts

中图分类号:

孙琪, 丛霞, 索佳佳, 曹荣峰, 姜忠玲, 崔凯, 高善颂, 田文儒. 钙／钙调素依赖性蛋白激酶Ⅱ对热休克小鼠胚胎成纤维细胞HSP70表达的影响[J]. 华北农学报, 2012, 27(4): 213-217. doi: 10.3969/j.issn.1000-7091.2012.04.041.

SUN Qi, CONG Xia, SUO Jia-jia, CAO Rong-feng, JIANG Zhong-ling, CUI Kai, GAO Shan-song, TIAN Wen-ru. Calcium/Calmodulin-dependent Protein Kinase Ⅱ Contributes to HSPTO Expression in Mouse Embryonic Fibroblasts[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2012, 27(4): 213-217. doi: 10.3969/j.issn.1000-7091.2012.04.041.

http://www.hbnxb.net/CN/Y2012/V27/I4/213

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