Growth factor mRNA and protein in preserved human amniotic membrane
Growth factor mRNA and protein in preserved human amniotic membrane
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DOI：
10.1076/0271-3683(200003)20:3;1-9;ft173
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发表时间：
2000-01-01
影响因子：
2
通讯作者：
Kinoshita, S
中科院分区：
文献类型：
--
作者：
Koizumi, N;Inatomi, T;Kinoshita, S
目的.目的：探讨生长因子mRNA在保存人羊膜中的表达及生长因子蛋白水平。采用RT-PCR方法检测人AM在-80 ℃保存1个月后8种生长因子（EGF、TGF-α、KGF、HGF、bFGF、TGF-β 1、TGF-β 2、TGF-β 3）和2种生长因子受体（KGFR和HGFR）mRNA的表达。此外，采用ELISA法检测了保存的人角膜和羊膜中7种生长因子（EGF、TGF-α、KGF、HGF、bFGF、TGF-β 1、TGF-β 2）的蛋白浓度。RT-PCR显示人AM表达EGF、TGF-α、KGF、HGF、bFGF、TGF-β 1、-β 2、-β 3、KGFR和HGFR的mRNA，而ELISA显示其含有EGF、TGF-α、KGF、HGF、bFGF、TGF-β 1、-β 2。没有羊膜上皮的AM也含有所有7种生长因子，然而，在该组织中，EGF，KGF，HGF和bFGF的蛋白水平被发现显着低于天然AM。保存的人AM表达许多生长因子的mRNA，并含有几种可能有利于AM移植后上皮形成的生长因子蛋白。高水平的EGF，KGF，HGF和碱性成纤维细胞生长因子与羊膜上皮AM相比，没有羊膜上皮表明上皮起源这些生长因子。我们认为EGF、KGF和HGF在AM移植后眼表创面愈合中可能起重要作用。
Purpose. To investigate the expression of growth factor mRNA and the level of growth factor protein in preserved human amniotic membrane (AM).Methods. RT-PCR was used to examine the expression of mRNA for eight growth factors (EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta 1, -beta 2, -beta 3) 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-beta 1, -beta 2) in preserved human corneas and in AM both with and without amniotic epithelium.Results. RT-PCR revealed that human AM expresses mRNA for EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta 1, -beta 2, -beta 3, KGFR and HGFR, while ELISAs showed that it contains EGF, TGF-alpha, KGF, HGF, bFGF, TGF-beta 1, -beta 2. 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.Conclusions. 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.