Kutty R K, Nagineni C N, Kutty G, Hooks J J, Chader G J, Wiggert B
Laboratory of Retinal Cell, National Eye Institute, National Institutes of Health, Bethesda, Maryland 20892.
J Cell Physiol. 1994 May;159(2):371-8. doi: 10.1002/jcp.1041590221.
Antibodies specific for heme oxygenase-1 (HO-1) were produced in rabbits, using the multiple antigen peptide (MAP) technique, and were employed to investigate the ability of transforming growth factor-beta 1 (TGF-beta 1) to induce the HO-1 protein in cultured human retinal pigment epithelial (RPE) cells. Western blot analyses showed that the cytokine induced HO-1 in these cells in a time- and dose-dependent manner. TGF-beta 1 also increased the mRNA for HO-1 in treated cells prior to the increase in HO-1 protein. The induction was effectively blocked by a neutralizing antibody preparation against TGF-beta 1. When tested under similar conditions, other growth factors such as basic fibroblast growth factor-I, platelet-derived growth factor, insulin-like growth factor, transforming growth factor-alpha, and epidermal growth factor did not show appreciable induction of HO-1. Lipopolysaccharide, tumor necrosis factor-alpha, and interferon-gamma were also not inducers, although TGF-beta 2 effectively induced HO-1. Heavy metal ions and thiol reagents were also highly potent inducers of HO-1 in human RPE cells. The induction of HO-1 by TGF-beta 1 was also observed in bovine choroid fibroblasts, but not in HELA, HEL or bovine corneal fibroblasts. Our results demonstrate for the first time that HO-1 can be induced by an important cytokine, TGF-beta 1, causing an increase in the expression of both HO-1 message and protein in specific neuroepithelial and fibroblast cells.
利用多抗原肽（MAP）技术在兔体内制备了血红素加氧酶-1（HO-1）特异性抗体，并用于研究转化生长因子-β1（TGF-β1）在培养的人视网膜色素上皮（RPE）细胞中诱导HO-1蛋白的能力。蛋白质印迹分析表明，该细胞因子以时间和剂量依赖性方式在这些细胞中诱导HO-1。在HO-1蛋白增加之前，TGF-β1还增加了处理细胞中HO-1的mRNA。针对TGF-β1的中和抗体制剂有效阻断了这种诱导作用。在相似条件下进行检测时，其他生长因子如碱性成纤维细胞生长因子-1、血小板衍生生长因子、胰岛素样生长因子、转化生长因子-α和表皮生长因子均未显示出明显的HO-1诱导作用。脂多糖、肿瘤坏死因子-α和干扰素-γ也不是诱导剂，尽管TGF-β2可有效诱导HO-1。重金属离子和硫醇试剂也是人RPE细胞中HO-1的高效诱导剂。在牛脉络膜成纤维细胞中也观察到TGF-β1对HO-1的诱导作用，但在HELA、HEL或牛角膜成纤维细胞中未观察到。我们的结果首次证明，HO-1可被重要的细胞因子TGF-β1诱导，导致特定神经上皮细胞和成纤维细胞中HO-1信息和蛋白的表达增加。
