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Abstract
BackgroundThe objective of this study was to evaluate the changes in gene expression after incubation of cells with proteins released from different silk mat layers.MethodsA silk cocoon from Bombyx mori was separated into four layers of equal thickness. The layers were numbered from 1 to 4 (from the inner to the outer layer). The proteins were released by sonication of a silk mat layer in normal saline. The concentration of proteins was determined by spectrophotometry. They were incubated with RAW264.7 cells, and changes in the expression of genes were evaluated by cDNA microarray analysis and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR).ResultsLayer 1 and 4 groups had higher protein concentrations compared to those in layer 2 and 3 groups. The genes associated with inflammation and angiogenesis showed significantly higher expression in layer 1 and 4 groups. The results of qRT-PCR were in agreement with those of the cDNA microarray analysis.ConclusionsThe silk mat from the middle portion of the silkworm cocoon yielded a lower protein release and caused an insignificant change in the expression of genes that are associated with inflammation and angiogenesis.
Highlights
The objective of this study was to evaluate the changes in gene expression after incubation of cells with proteins released from different silk mat layers
The genes in the transforming growth factor-β (TGF-β) family and the genes associated with inflammation showed higher levels of expression in layer 1 and 4 groups than in layer 2 and 3 groups
In this study, a gene expression profile of macrophages was evaluated by cDNA microarray and quantitative reverse transcriptase-polymerase chain reaction (qRT-PCR) analyses
Summary
The objective of this study was to evaluate the changes in gene expression after incubation of cells with proteins released from different silk mat layers. Because the silkworm cocoon is mainly composed of proteins fibroin and sericin, the silk mat has composition similar to that of the silkworm cocoon [3]. Sericin content generally increases from the inner layer to the outer layer [5]. Aside from these two proteins, seroin and protease inhibitors can be found in the silk mat [4]. The concentrations of these proteins are different among layers. Seroin and several protease inhibitors are enriched in the innermost layer [4]. There has been few studies to illuminate these differences
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