Differential expression and regulation of TGF-β1, TGF-β2, TGF-β3, TGF-βRI, TGF-βRII and TGF-βRIII in cultured human corneal, limbal, and conjunctival fibroblasts
Differential expression and regulation of TGF-β1, TGF-β2, TGF-β3, TGF-βRI, TGF-βRII and TGF-βRIII in cultured human corneal, limbal, and conjunctival fibroblasts
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DOI：
10.1076/ceyr.19.2.154.5321
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发表时间：
1999-08-01
影响因子：
2
通讯作者：
Tseng, SCG
中科院分区：
文献类型：
--
作者：
Li, DQ;Lee, SB;Tseng, SCG
目的。我们报道了人眼表面上皮细胞和成纤维细胞之间可能涉及细胞因子表达的三种模式。 TGF-β家族是一种典型的纤维形成细胞因子，负责伤口愈合中成纤维细胞的活化。我们研究了 TGF-β 家族在培养的人角膜、角膜缘和结膜成纤维细胞中如何差异表达和调节。方法。将人角膜 (HCF)、角膜缘 (HLF) 和结膜成纤维细胞 (HJF) 在 DMEM-10% FBS 中培养直至汇合，然后切换至无血清 DMEM-ITS 48 小时，然后在三个独立实验中添加 8 种细胞因子中的每一种 10 ng/ml 4 小时。分离总RNA并以GAPDH作为对照进行Northern杂交。使用ELISA测定培养基中的TGF-β1和TGF-β2蛋白。结果。 HCF、HLF 和 HJF 均表达 TGF-β 的所有三种同工型和 TGF-β R 的三种类型。在所有三种类型的成纤维细胞中，TGF-β 1 mRNA 的表达最强，并被三种 TGF-β 上调。 PDGF-BB 和 TGF-α 略微增加 TGF-β 1 mRNA。 TGF-β 还上调 HJF 中的 TGF-β 3 mRNA。 TGF-β RI mRNA 是唯一被 TGF-β s 上调的受体。 TGF-β RII 和 TGF-β RIII mRNA 并非受所有测试的细胞因子调节。结论。 TGF-β的自诱导是上调TGF-β1表达的主要机制。 TGF-β 促进 TGF-β 3 可能在 HJF 中具有特殊作用。 TGF-β s 和 TGF-β Rs 的差异表达和调节表明每种 TGF-β 亚型可能在不同的眼表成纤维细胞中具有特定的功能。没有测试的细胞因子可以下调 TGF-β 1 和 TCF-β Rs。
Purpose. We have reported that three patterns of cytokine expression are potentially involved between epithelia and fibroblasts of the human ocular surface. The TGF-beta family is a prototypical fibrogenic cytokine responsible for fibroblast activation in wound healing. We investigated how the TGF-beta family is differentially expressed and regulated in cultured human corneal, limbal and conjunctival fibroblasts.Methods. Human corneal (HCF), limbal (HLF) and conjunctival fibroblast (HJF) were cultured in DMEM-10% FBS until confluence and switched to serum-free DMEM-ITS for 48 h before adding 10 ng/ml of each of eight cytokines for 4 h in three separate experiments. Total RNA was isolated and subjected to Northern hybridization with GAPDH as a control. ELISA was used to determine TGF-beta 1 and TGF-beta 2 proteins in the media.Results. All three isoforms of TGF-beta and three types of TGF-beta R were expressed by HCF, HLF and HJF. Expression of TGF-beta 1 mRNA was strongest and upregulated by the three TGF-beta s in all three types of fibroblast. PDGF-BB and TGF-alpha slightly increased TGF-beta 1 mRNA. TGF-beta s also upregulated TGF-beta 3 mRNA in HJF. TGF-beta RI mRNA was the only receptor upregulated by TGF-beta s. TGF-beta RII and TGF-beta RIII mRNA were not regulated by all cytokines tested.Conclusions. TGF-beta s auto-induction is the major mechanism upregulating TGF-beta 1 expression. Promotion of TGF-beta 3 by the TGF-beta s may have a special role in HJF. Differential expression and regulation of TGF-beta s and TGF-beta Rs suggest that each TGF-beta isoform may have specific functions in different ocular surface fibroblasts. No cytokine tested can downregulate TGF-beta 1 and the TCF-beta Rs.