Koizumi N J, Inatomi T J, Sotozono C J, Fullwood N J, Quantock A J, Kinoshita S
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Japan.
Curr Eye Res. 2000 Mar;20(3):173-7.
To investigate the expression of growth factor mRNA and the level of growth factor protein in preserved human amniotic membrane (AM).
RT-PCR was used to examine the expression of mRNA for eight growth factors (EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta1, -beta2, -beta3) and two growth factor receptors (KGFR and HGFR) in human AM preserved at -80 degrees C for one month. In addition, ELISAs were used to measure the protein concentrations of seven growth factors (EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta1, -beta2) in preserved human corneas and in AM both with and without amniotic epithelium.
RT-PCR revealed that human AM expresses mRNA for EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta1, -beta2, -beta3, KGFR and HGFR, while ELISAs showed that it contains EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta1, -beta2. AM without amniotic epithelium also contains all seven growth factors examined, however, in this tissue the protein levels of EGF, KGF, HGF and bFGF were found to be significantly lower than in native AM.
Preserved human AM expresses mRNAs for a number of growth factors and contains several growth factor proteins that might benefit epithelialization after AM transplantation. High levels of EGF, KGF, HGF and bFGF in AM with amniotic epithelium as compared to AM without amniotic epithelium suggest an epithelial origin for these growth factors. We feel that EGF, KGF and HGF in particular might play important roles in ocular surface wound healing after AM transplantation.
研究保存的人羊膜（AM）中生长因子mRNA的表达及生长因子蛋白水平。
采用逆转录-聚合酶链反应（RT-PCR）检测在-80℃保存1个月的人羊膜中8种生长因子（表皮生长因子[EGF]、转化生长因子-α[TGF-α]、角质形成细胞生长因子[KGF]、肝细胞生长因子[HGF]、碱性成纤维细胞生长因子[bFGF]、转化生长因子-β1、-β2、-β3）和2种生长因子受体（角质形成细胞生长因子受体[KGFR]和肝细胞生长因子受体[HGFR]）的mRNA表达。此外，采用酶联免疫吸附测定（ELISA）法检测保存的人角膜以及有和没有羊膜上皮的羊膜中7种生长因子（EGF、TGF-α、KGF、HGF、bFGF、TGF-β1、-β2）的蛋白浓度。
RT-PCR显示人羊膜表达EGF、TGF-α、KGF、HGF、bFGF、TGF-β1、-β2、-β3、KGFR和HGFR的mRNA，而ELISA显示其含有EGF、TGF-α、KGF、HGF、bFGF、TGF-β1、-β2。没有羊膜上皮的羊膜也含有所检测的全部7种生长因子，然而，在该组织中，EGF、KGF、HGF和bFGF的蛋白水平显著低于天然羊膜。
保存的人羊膜表达多种生长因子的mRNA，并含有几种生长因子蛋白，这些蛋白可能有利于羊膜移植后的上皮形成。与没有羊膜上皮的羊膜相比，有羊膜上皮的羊膜中高水平的EGF、KGF、HGF和bFGF提示这些生长因子起源于上皮。我们认为，尤其是EGF、KGF和HGF可能在羊膜移植后的眼表伤口愈合中发挥重要作用。
