European Center for Angioscience (ECAS), Medical Faculty Mannheim of the University of Heidelberg, Ludolf-Krehl-Strasse 13-17, 68167, Mannheim, Germany.
Mannheim Institute for Innate Immunoscience (MI3), Medical Faculty Mannheim of the University of Heidelberg, Ludolf-Krehl-Strasse 13-17, 68167, Mannheim, Germany.
Sci Rep. 2023 Oct 20;13(1):17985. doi: 10.1038/s41598-023-45139-7.
In diabetic nephropathy (DN), glomerular endothelial cells (GECs) and podocytes undergo pathological alterations, which are influenced by metabolic changes characteristic of diabetes, including hyperglycaemia (HG) and elevated methylglyoxal (MGO) levels. However, it remains insufficiently understood what effects these metabolic factors have on GEC and podocytes and to what extent the interactions between the two cell types can modulate these effects. To address these questions, we established a co-culture system in which GECs and podocytes were grown together in close proximity, and assessed transcriptional changes in each cell type after exposure to HG and MGO. We found that HG and MGO had distinct effects on gene expression and that the effect of each treatment was markedly different between GECs and podocytes. HG treatment led to upregulation of "immediate early response" genes, particularly those of the EGR family, as well as genes involved in inflammatory responses (in GECs) or DNA replication/cell cycle (in podocytes). Interestingly, both HG and MGO led to downregulation of genes related to extracellular matrix organisation in podocytes. Crucially, the transcriptional responses of GECs and podocytes were dependent on their interaction with each other, as many of the prominently regulated genes in co-culture of the two cell types were not significantly changed when monocultures of the cells were exposed to the same stimuli. Finally, the changes in the expression of selected genes were validated in BTBR ob/ob mice, an established model of DN. This work highlights the molecular alterations in GECs and podocytes in response to the key diabetic metabolic triggers HG and MGO, as well as the central role of GEC-podocyte crosstalk in governing these responses.
在糖尿病肾病 (DN) 中，肾小球内皮细胞 (GEC) 和足细胞发生病理改变，这些改变受糖尿病特征性代谢变化的影响，包括高血糖 (HG) 和甲基乙二醛 (MGO) 水平升高。然而，人们对这些代谢因素对 GEC 和足细胞的影响以及这两种细胞类型之间的相互作用在多大程度上可以调节这些影响知之甚少。为了解决这些问题，我们建立了一种共培养系统，其中 GEC 和足细胞紧密地生长在一起，并在暴露于 HG 和 MGO 后评估每种细胞类型的转录变化。我们发现 HG 和 MGO 对基因表达有不同的影响，每种处理对 GEC 和足细胞的影响明显不同。HG 处理导致“即刻早期反应”基因上调，特别是 EGR 家族的基因，以及参与炎症反应（在 GEC 中）或 DNA 复制/细胞周期（在足细胞中）的基因上调。有趣的是，HG 和 MGO 处理都导致足细胞中与细胞外基质组织相关的基因下调。至关重要的是，GEC 和足细胞的转录反应依赖于它们之间的相互作用，因为当两种细胞的单核培养物暴露于相同的刺激时，共培养的两种细胞中许多明显调节的基因没有显著变化。最后，在 BTBR ob/ob 小鼠中验证了选定基因表达的变化，BTBR ob/ob 小鼠是一种已建立的 DN 模型。这项工作强调了 GEC 和足细胞对关键糖尿病代谢触发物 HG 和 MGO 的分子改变，以及 GEC-足细胞串扰在调节这些反应中的核心作用。
