Regenerative Medicine Institute (REMEDI) at CÚRAM Centre for Research in Medical Devices, School of Medicine, National University of Ireland Galway, Galway, REMEDI, Biomedical Sciences, Corrib Village, Dangan, Galway, H91 TK33, Ireland.
Centre for Endocrinology, Diabetes and Metabolism, Galway University Hospitals, Galway, Ireland.
Stem Cell Res Ther. 2019 Nov 19;10(1):329. doi: 10.1186/s13287-019-1424-5.
Renal proximal tubular epithelial cells (RPTEC) are dysfunctional in diabetic kidney disease (DKD). Mesenchymal stromal cells (MSC) may modulate DKD pathogenesis through anti-inflammatory mediators. This study aimed to investigate the pro-inflammatory effect of extended exposure to high glucose (HG) concentration on stable RPTEC monolayers and the influence of MSC on this response.
Morphologically stable human RPTEC/TERT1 cell monolayers were exposed to 5 mM and 30 mM (HG) D-glucose or to 5 mM D-glucose + 25 mM D-mannitol (MAN) for 5 days with sequential immunoassays of supernatants and end-point transcriptomic analysis by RNA sequencing. Under the same conditions, MSC-conditioned media (MSC-CM) or MSC-containing transwells were added for days 4-5. Effects of CM from HG- and MAN-exposed RPTEC/MSC co-cultures on cytokine secretion by monocyte-derived macrophages were determined.
After 72-80 h, HG resulted in increased RPTEC/TERT1 release of interleukin (IL)-6, IL-8, monocyte chemoattractant protein (MCP)-1 and neutrophil gelatinase-associated lipocalin (NGAL). The HG pro-inflammatory effect was attenuated by concentrated (10×) MSC-CM and, to a greater extent, by MSC transwell co-culture. Bioinformatics analysis of RNA sequencing data confirmed a predominant effect of HG on inflammation-related mediators and biological processes/KEGG pathways in RPTEC/TERT1 stable monolayers as well as the non-contact-dependent anti-inflammatory effect of MSC. Finally, CM from HG-exposed RPTEC/MSC transwell co-cultures was associated with attenuated secretion of inflammatory mediators by macrophages compared to CM from HG-stimulated RPTEC alone.
Stable RPTEC monolayers demonstrate delayed pro-inflammatory response to HG that is attenuated by close proximity to human MSC. In DKD, this MSC effect has potential to modulate hyperglycemia-associated RPTEC/macrophage cross-talk.
在糖尿病肾病（DKD）中，肾近端管状上皮细胞（RPTEC）功能失调。间充质基质细胞（MSC）可能通过抗炎介质来调节 DKD 的发病机制。本研究旨在探讨高浓度葡萄糖（HG）长期暴露对稳定 RPTEC 单层的促炎作用，以及 MSC 对这种反应的影响。
用形态稳定的人 RPTEC/TERT1 细胞单层分别暴露于 5 mM 和 30 mM（HG）D-葡萄糖或 5 mM D-葡萄糖+25 mM D-甘露醇（MAN）5 天，并用上清液的连续免疫测定和终点 RNA 测序的转录组分析。在相同条件下，在第 4-5 天加入 MSC 条件培养基（MSC-CM）或含有 MSC 的 Transwell。确定 HG 和 MAN 暴露的 RPTEC/MSC 共培养物的 CM 对单核细胞来源的巨噬细胞细胞因子分泌的影响。
在 72-80 小时后，HG 导致 RPTEC/TERT1 释放白细胞介素（IL）-6、IL-8、单核细胞趋化蛋白（MCP）-1 和中性粒细胞明胶酶相关脂质运载蛋白（NGAL）增加。浓缩（10×）MSC-CM 以及更显著的 MSC Transwell 共培养减轻了 HG 的促炎作用。RNA 测序数据的生物信息学分析证实，HG 对 RPTEC/TERT1 稳定单层中的炎症相关介质和生物学过程/KEGG 途径有主要影响，并且 MSC 具有非接触依赖性抗炎作用。最后，与仅 HG 刺激的 RPTEC 相比，来自 HG 暴露的 RPTEC/MSC Transwell 共培养物的 CM 与炎症介质分泌减少相关。
稳定的 RPTEC 单层对 HG 表现出延迟的促炎反应，而与人类 MSC 的近距离接近则减轻了这种反应。在 DKD 中，MSC 效应有可能调节高血糖相关的 RPTEC/巨噬细胞相互作用。
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