Suppression of TGF-ß signaling in both normal conjunctival fibroblasts and pterygial body fibroblasts by amniotic membrane
Suppression of TGF-ß signaling in both normal conjunctival fibroblasts and pterygial body fibroblasts by amniotic membrane
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DOI：
10.1076/0271-3683(200004)2041-5ft325
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发表时间：
2000-01
影响因子：
2
通讯作者：
Sao-Bing Lee;De-Quan Li;D. Tan;D. Meller;S. Tseng
中科院分区：
文献类型：
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作者：
Sao-Bing Lee;De-Quan Li;D. Tan;D. Meller;S. Tseng
目的.在翼状胬肉和其他眼表疾病的裸巩膜切除术后，羊膜移植可作为替代基质，减少重建结膜表面的瘢痕形成。本研究旨在确定羊膜（AM）是否抑制培养的正常结膜（HCF）和结膜体成纤维细胞（PBF）中TGF β信号系统的表达。方法. HCF和PBF在AM和塑料威尔斯孔中在含或不含TGF-β 1的含血清和无血清DMEM中培养。提取总RNA，用TGF-β 1、β 2和β 3; TGF-β 1受体（TGF-β R）I、II和III型; α-平滑肌肌动蛋白（α-SM）、β 1-整联蛋白、CD 44、成纤维细胞生长因子受体1（FGF-R1/β）和血小板衍生生长因子受体β 1（PDGFR-R1）;和GAPDH作为上样对照的探针进行北方杂交。MTT法检测细胞增殖。结果与塑料表面培养物相比，羊膜显著抑制两种成纤维细胞的TGF-β 2、β 3和所有三种类型的TGF-β受体的转录表达。此外，CD 44转录物的表达也被显著抑制，而β 1整联蛋白、α-SM肌动蛋白和FGFR 1/β 1的表达被轻度抑制。相反，TGF-β 1和PDGFR-β 1的表达基本保持不变。AM上生长的HCF和PBF的细胞增殖也受到明显抑制。结论.羊膜基质独特地抑制两种类型的成纤维细胞中的TGF-β信号传导。它还可以抑制通过CD44、β 1整联蛋白和FGFR1/β 2的信号传导。因此，表型可能变得不那么促有丝分裂、收缩和纤维化。这些数据部分支持了羊膜移植对结膜表面重建具有抗瘢痕形成作用的原因。
Purpose. When used as an alternative substrate following bare sclera removal of pterygium and other ocular surface diseases, amniotic membrane transplantation can reduce scarring on the reconstructed conjunctival surface. This study was carried out to determine if the amniotic membrane (AM) suppresses the expression of the TGFß signaling system in cultured normal conjunctival (HCF) and pterygial body fibroblasts (PBF). Methods. HCF and PBF were cultured on AM and plastic wells in serum-containing and serum-free DMEM with or without TGF-ß1. Total RNA was extracted and subjected to Northern hybridization with probes of TGF-ß1, ß2 and ß3; TGF-ß receptors (TGF-ßR) type I, II and III; a-smooth muscle actin (a-SM), ß1-integrin, CD44, fibroblast growth factor receptor 1 (FGF-R1/ flg) and platelet-derived growth factor receptor ß (PDGFR-ß); and GAPDH as a loading control. MTT assay was used for cell proliferation. Results. Amniotic membrane markedly suppressed the transcript expression of TGF-ß2, ß3 and all three types of TGF-ß receptors by both fibroblasts as compared to their cultures on plastic surface. In addition, expression of CD44 transcript was also markedly suppressed while that of ß1 integrin, a-SM actin, and FGFR1/ flg was mildly suppressed. In contrast, expression of TGF-ß1 and PDGFR-ß remained largely unchanged. The cell proliferation of HCF and PBF grown on AM was also significantly suppressed. Conclusions. Amniotic membrane matrix uniquely suppresses TGF-ß signaling in both types of fibroblasts. It may also suppress signaling via CD44, ß1 integrin and FGFR1/ flg. As a result, the phenotype may become less mitogenic, contractile and fibrogenic. These data support in part why amniotic membrane transplantation has an anti-scarring effect for conjunctival surface reconstruction.