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Immunohistochemical and transcriptional expression of Matrix Metalloproteinases in full-term human umbilical cord and Human Umbilical Vein Endothelial Cells

机译:基质金属蛋白酶在足月人脐带和人脐静脉内皮细胞中的免疫组织化学和转录表达

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摘要

Matrix metalloproteinases (MMPs) are extracellular zinc-dependent endopeptidases involved in the degradation and remodelling of extracellular matrix in physiological and pathological processes. MMPs also have a role on cell proliferation, migration, differentiation, angiogenesis and apoptosis. Umbilical cord is a special organ subjected to many changes during pre-natal life and whose cells can maintain a certain degree of plasticity also in post-natal period; for example recently they have been used as a source of stem cells. udIn this work we investigated the expression of MMPs in human umbilical cord and Human Umbilical Vein Endothelial Cells (HUVEC) though immunohistochemistry, RT-PCR and gelatin zymography. MMP-2 protein is expressed in the amniotic epithelium of human umbilical cord and in few sub-epithelial fibroblasts, while MMP-3 and MMP-10 only in the umbilical epithelium. MMP-8, MMP-9 and MMP-13 immunoreactivity is localised in the epithelium and in Wharton’s jelly mesenchymal cells. Immunocytochemistry also revealed protein expression for MMP-2, 3, 8, 9 and 10 in cultured HUVEC.udIn agreement with immunohistochemical data, RT-PCR analysis performed on samples of whole umbilical cord confirmed the transcriptional expression for the genes encoding all the six matrix metalloproteinases investigated, while in HUVEC only the expression of MMP-2, 3, 9, 10 and 13 mRNAs was detected. udGelatin zymograpgy showed a clear MMP-2 and MMP-9 enzymatic activity in the conditioned medium of HUVEC at different culture passages, suggesting that HUVEC secrete gelatinases, that afterwards undergo extracellular activation, and this ability is not affected by passage number.
机译:基质金属蛋白酶(MMPs)是细胞外锌依赖性内肽酶,参与生理和病理过程中细胞外基质的降解和重塑。 MMPs也对细胞增殖,迁移,分化,血管生成和凋亡具有作用。脐带是一种特殊的器官,在产前生活中会发生许多变化,其细胞在产后也可以保持一定程度的可塑性。例如,最近它们已被用作干细胞的来源。 ud在这项工作中,我们通过免疫组织化学,RT-PCR和明胶酶谱分析研究了MMPs在人脐带和人脐静脉内皮细胞(HUVEC)中的表达。 MMP-2蛋白在人脐带的羊膜上皮和很少的上皮下成纤维细胞中表达,而MMP-3和MMP-10仅在脐带上皮中表达。 MMP-8,MMP-9和MMP-13的免疫反应性位于上皮和沃顿的果冻间充质细胞中。免疫细胞化学法还显示了培养的HUVEC中MMP-2、3、8、9和10的蛋白质表达。 ud与免疫组织化学数据一致,对全脐带样品进行的RT-PCR分析证实了编码所有这6种基因的基因的转录表达研究了基质金属蛋白酶,而在HUVEC中仅检测到MMP-2、3、9、10和13 mRNA的表达。明胶酶解法在不同培养传代的HUVEC条件培养基中显示出清晰的MMP-2和MMP-9酶促活性,表明HUVEC分泌明胶酶,此后再经历细胞外激活,并且该能力不受传代次数的影响。

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