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Cortisol Effect on Heat Shock Proteins in the C2C12 and 3T3-L1 Cells

机译:皮质醇对C2C12和3T3-L1细胞中热休克蛋白的影响

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摘要

The present study was carried out to understand the effect of cortisol on heat shock protein system (Hsps) in the C2C12 and 3T3-L1 cells under co-culture system. Cells were co-cultured by using Transwell inserts with a 0.4-mu m porous membrane to separate C2C12 and 3T3-L1 cells. Each cell type was grown independently on the Transwell plates. After cell differentiation, inserts containing 3T3-L1 cells were transferred to C2C12 plates and inserts containing C2C12 cells transferred to 3T3-L1 plates. Ten micrograms per microliter of cortisol was added to the medium. Following 72 h of treatment, the cells in the lower wells were harvested for analysis. Heat shock proteins (Hsps) such as Hsp27, Hsp70, and Hsp90 were selected for the analysis. The qRT-PCR results showed the significant increase in the mRNA expression of as Hsp27, Hsp70, and Hsp90. In addition, confocal microscopical investigation showed the cortisol treatment increases Hsps expressions in the mono and co-cultured C2C12 and 3T3-L1 cells. From the results, we concluded that the cortisol increases Hsps expression in the co-cultured C2C12 and 3T3-L1 cells, which is differed from one-dimensional mono-cultured C2C12 and 3T3-L1 cells.
机译:本研究旨在了解皮质醇对共培养系统下C2C12和3T3-L1细胞中热休克蛋白系统(Hsp​​s)的影响。通过使用带有0.4微米多孔膜的Transwell插入物共培养细胞,以分离C2C12和3T3-L1细胞。每种细胞类型均在Transwell板上独立生长。细胞分化后,将含有3T3-L1细胞的插入片段转移到C2C12平板上,将含有C2C12细胞的插入片段转移到3T3-L1平板上。每微升皮质醇十微克被加入到培养基中。处理72小时后,收集下部孔中的细胞用于分析。选择热休克蛋白(Hsps)(例如Hsp27,Hsp70和Hsp90)进行分析。 qRT-PCR结果显示as Hsp27,Hsp70和Hsp90的mRNA表达显着增加。另外,共聚焦显微镜研究表明,皮质醇处理增加了单培养和共培养的C2C12和3T3-L1细胞中Hsps的表达。从结果可以得出结论,皮质醇增加了共培养的C2C12和3T3-L1细胞中Hsps的表达,这与一维单培养的C2C12和3T3-L1细胞不同。

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