Wang Y, Xiao X, Huang S, Luo F, Luo H, You J, Luo Z
Department of Pathophysiology, Hunan Medical University, Changsha.
Hunan Yi Ke Da Xue Xue Bao. 1997;22(1):1-4.
The aim of this study is to determine whether heat-induced heat shock gene expression in rat pulmonary alveolar macrophages (PAMs) would protect PAMs against hydrogen peroxide (H2O2)-mediated cell killing. In response to sublethal heat shock at 42 degrees C for 2 h, the cells synthesized heat shock protein 70 kD (HSP70) and other different molecular weight heat shock proteins, which were detected with gel electrophoresis after [35S]-methionin labeling cellular protein and Western blotting analysis in PAMs. Northern blot analysis showed the induction of HSP70 mRNA with heat shock treatment. Futhermore, with heat shock, there was a significant increment of survival cells after H2O2 (1,2,3 mmol.L-1, 45 min) exposure. This increase was blocked both by Cycloheximide and by Actinomycin D, and was associated with inhibition of synthesis of HSP70 protein and transcription of HSP70 mRNA. These results strongly suggest that expression of heat shock gene (especially HSP70 gene) plays an important role in the intracelluar mechanism of cytoprotection against H2O2 in rat PAMs.
本研究的目的是确定大鼠肺泡巨噬细胞(PAMs)中热诱导的热休克基因表达是否能保护PAMs免受过氧化氢(H2O2)介导的细胞杀伤。在42℃下进行亚致死性热休克处理2小时后,细胞合成了70kD热休克蛋白(HSP70)和其他不同分子量的热休克蛋白,在对PAMs细胞蛋白进行[35S] - 蛋氨酸标记并用凝胶电泳和蛋白质免疫印迹分析后检测到这些蛋白。Northern印迹分析显示热休克处理可诱导HSP70 mRNA。此外,热休克处理后,H2O2(1、2、3 mmol.L-1,45分钟)暴露后的存活细胞显著增加。这种增加被放线菌酮和放线菌素D阻断,并且与HSP70蛋白合成的抑制和HSP70 mRNA转录的抑制有关。这些结果强烈表明热休克基因(尤其是HSP70基因)的表达在大鼠PAMs细胞内对抗H2O2的细胞保护机制中起重要作用。