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首页> 外文期刊>American Journal of Physiology >Functional role of bone morphogenetic protein-4 in isolated canine parietal cells.
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Functional role of bone morphogenetic protein-4 in isolated canine parietal cells.

机译：骨形态发生蛋白4在孤立的犬壁细胞中的功能作用。

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摘要

Bone morphogenetic protein (BMP)-4 is an important regulator of cellular growth and differentiation. Expression of BMP-4 has been documented in the gastric mucosa. We reported that incubation of canine parietal cells with EGF for 72 h induced both parietal cell morphological transformation and inhibition of H(+)/K(+)-ATPase gene expression through MAPK-dependent mechanisms. We explored the role of BMP-4 in parietal cell maturation and differentiation. Moreover, we investigated if BMP-4 modulates the actions of EGF in parietal cells. H(+)/K(+)-ATPase gene expression was examined by Northern blots and quantitative RT-PCR. Acid production was assessed by measuring the uptake of [(14)C]aminopyrine. Parietal cell apoptosis was quantitated by Western blots with anti-cleaved caspase 3 antibodies and by counting the numbers of fragmented, propidium iodide-stained nuclei. MAPK activation and Smad1 phosphorylation were measured by Western blots with anti-phospho-MAPK and anti-phospho-Smad1 antibodies. Parietal cell morphology was examined by immunohistochemical staining of cells with anti-H(+)/K(+)-ATPase alpha-subunit antibodies. BMP-4 stimulated Smad1 phosphorylation and induced H(+)/K(+)-ATPase gene expression. BMP-4 attenuated EGF-mediated inhibition of H(+)/K(+)-ATPase gene expression and blocked EGF induction of both parietal cell morphological transformation and MAPK activation. Incubation of cells with BMP-4 enhanced histamine-stimulated [(14)C]aminopyrine uptake. BMP-4 had no effect on parietal cell apoptosis, whereas TGF-beta stimulated caspase-3 activation and nuclear fragmentation. In conclusion, BMP-4 promotes the induction and maintenance of a differentiated parietal cell phenotype. These findings may provide new clues for a better understanding of the mechanisms that regulate gastric epithelial cell growth and differentiation.
机译：骨形态发生蛋白（BMP）-4是细胞生长和分化的重要调节剂。 BMP-4的表达已在胃粘膜中被证明。我们报告说，用EGF孵育犬壁细胞72小时，可通过MAPK依赖性机制诱导壁细胞形态转化和H（+）/ K（+）-ATPase基因表达的抑制。我们探讨了BMP-4在壁细胞成熟和分化中的作用。此外，我们调查了BMP-4是否调节壁细胞中EGF的作用。 H（+）/ K（+）-ATPase基因表达通过Northern印迹和定量RT-PCR检查。通过测量[（14）C]氨基比林的摄入量来评估产酸量。通过用抗裂解的caspase 3抗体进行Western印迹，并通过计数碘化丙锭染色的碎裂核的数量，定量壁细胞的凋亡。 MAPK活化和Smad1磷酸化通过Western blot检测抗磷酸化MAPK和抗磷酸化Smad1抗体。通过用抗H（+）/ K（+）-ATPaseα-亚基抗体对细胞进行免疫组织化学染色来检查壁细胞的形态。 BMP-4刺激Smad1磷酸化并诱导H（+）/ K（+）-ATPase基因表达。 BMP-4减弱了EGF介导的H（+）/ K（+）-ATPase基因表达的抑制作用，并阻断了EGF诱导的壁细胞形态转化和MAPK激活。用BMP-4孵育细胞可增强组胺刺激的[（14）C]氨基比林吸收。 BMP-4对壁细胞凋亡无影响，而TGF-β刺激caspase-3激活和核碎裂。总之，BMP-4促进分化的壁细胞表型的诱导和维持。这些发现可能为更好地理解调节胃上皮细胞生长和分化的机制提供新的线索。

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