J Korean Ophthalmol Soc.  2001 Dec;42(12):1760-1770.

The Differential Gene Expression in Corneal Fibroblast Cultured with Amniotic Membrane Components

• Kim YJ
• Shyn KH
• Kim JC

Abstract

PURPOSE: To know whether amniotic membrane (AM) extract could protect cell from toxic stimuli by changing gene expression such as inflammatory and apoptosis-related genes, cultured corneal fibroblasts were used.
METHODS
After AM was pulverized in the liquid nitrogen and homogenized under the 4(C, it was centrifuged and its supernatant was obtained. Human corneal fibroblasts (HCFs) were divided into 2 groups - one group treated with AM extracts and TNF-alpha, and the other with TNF-alpha only. Cell morphology and viability were assessed and differential gene expression through cDNA array was performed.
RESULTS
Viability of HCF cultured in AM extracts and TNF-alpha was better than that of cells in TNF-alpha only. Cells in TNF-alpha only were morphologically changed and dead. cDNA array showed the decrease in IL-1 , IL-1 receptor type 1 and caspases-1,-3,and -10 expression in cells treated with AM extracts. MMP-1,-2,-3 and-9 were almost the same or slightly increased and NF-kappaB, I kappaB kinase alpha, integrin-alpha1 and -alpha10, and MAP kinase were increased in cells with AM extracts.
CONCLUSIONS
When cultured corneal fibroblasts were insulted by TNF-alpha, AM extracts seemed to protect the cells, by down-regulating various inflammation - and cell death- related genes, and by increasing the expression of genes enhancing cell proliferation and differentiation.

Keyword

Amniotic membrane; Corneal fibroblast; cDNA array; Differential gene expression

MeSH Terms

Amnion*
Cell Proliferation
Fibroblasts*
Gene Expression*
Humans
Inflammation
Interleukin-1
NF-kappa B
Nitrogen
Oligonucleotide Array Sequence Analysis
Phosphotransferases
Tumor Necrosis Factor-alpha
Interleukin-1
NF-kappa B
Nitrogen
Phosphotransferases
Tumor Necrosis Factor-alpha