Cowin A J, Bidey S P
Department of Medicine, University of Manchester, UK.
J Endocrinol. 1995 Jan;144(1):67-73. doi: 10.1677/joe.0.1440067.
The release of latent transforming growth factor-beta 1 (TGF-beta 1), and conversion to the biologically active peptide, has been investigated in porcine thyroid follicular cells maintained in primary monolayer culture. Analysis by radioreceptor assay of medium conditioned for 72 h by subconfluent thyroid monolayers showed that a high proportion of the expressed TGF-beta 1 peptide was in the active form. Medium conditioned by iodide (10 mumol/l)-treated follicular cells contained higher levels of both active and total TGF-beta 1 than were present in medium conditioned by untreated cells. Exposure of cells to iodide also led to a marked decrease in [methyl-3H]thymidine incorporation that was relieved by immunoadsorption with a neutralizing antiserum against the active form of TGF-beta 1. Inclusion of a low dose (80 units/l) of porcine plasmin led to a small increase in incorporation of [methyl-3H]thymidine, while higher doses of plasmin (1250-5000 units/l) or plasminogen (100 mg/l) significantly reduced [methyl-3H]thymidine incorporation. This inhibition was effectively reversed by immunoadsorption of TGF-beta 1 from the medium during the test incubations. The study therefore provides direct evidence for a stimulatory role of thyroidal iodide in enhancing the release of latent TGF-beta 1 peptide, and suggests that in normal thyroid follicular cells, as in other TGF-beta 1 producing epithelia, post-secretory processing to the biologically active molecule occurs through an endogenous cellular mechanism. It appears likely that plasmin, generated locally within the thyroid follicular microenvironment, may play a fundamental role in effecting this conversion.
在原代单层培养的猪甲状腺滤泡细胞中,对潜伏转化生长因子-β1(TGF-β1)的释放及其向生物活性肽的转化进行了研究。通过放射受体分析对亚汇合甲状腺单层细胞培养72小时的条件培养基进行分析,结果表明,表达的TGF-β1肽中有很大一部分是活性形式。经碘化物(10μmol/L)处理的滤泡细胞的条件培养基中,活性TGF-β1和总TGF-β1的水平均高于未处理细胞的条件培养基。细胞暴露于碘化物还导致[甲基-3H]胸腺嘧啶核苷掺入量显著降低,而用针对活性形式TGF-β1的中和抗血清进行免疫吸附可缓解这种降低。加入低剂量(80单位/L)的猪纤溶酶会使[甲基-3H]胸腺嘧啶核苷掺入量略有增加,而高剂量的纤溶酶(1250 - 5000单位/L)或纤溶酶原(100mg/L)则显著降低[甲基-3H]胸腺嘧啶核苷掺入量。在测试孵育期间,通过从培养基中免疫吸附TGF-β1可有效逆转这种抑制作用。因此,该研究为甲状腺碘化物在增强潜伏TGF-β1肽释放中的刺激作用提供了直接证据,并表明在正常甲状腺滤泡细胞中,与其他产生TGF-β1的上皮细胞一样,向生物活性分子的分泌后加工是通过内源性细胞机制发生的。甲状腺滤泡微环境中局部产生的纤溶酶似乎可能在实现这种转化中起重要作用。