Department of nephrology, Jiangsu Province Hospital of Chinese Medicine, The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China,; Key Laboratory for Metabolic Diseases in Chinese Medicine, First College of Clinical Medicine, Nanjing University of Chinese Medicine, Nanjing 210023, China.
Department of nephrology, Jiangsu Province Hospital of Chinese Medicine, The Affiliated Hospital of Nanjing University of Chinese Medicine, Nanjing 210029, China.
Phytomedicine. 2024 Jul;129:155683. doi: 10.1016/j.phymed.2024.155683. Epub 2024 Apr 25.
Peritoneal dialysis (PD) is a successful renal replacement therapy for end-stage renal disease. Long-term PD causes mesothelial-mesenchymal transition (MMT) of peritoneal mesothelial cells (PMCs), leading to peritoneal fibrosis (PF), which reduces the efficiency of PD. Macrophages are thought to play a role in the onset and perpetuation of peritoneal injury. However, the mechanisms by which macrophages-PMCs communication regulates peritoneal fibrosis are not fully understood resulting in a lack of disease-modifying drugs. Astragaloside IV (AS-IV) possessed anti-fibrotic effect towards PF in PD whereas the mechanistic effect of AS-IV in PD is unknown.
The primary macrophages were extracted and treated with LPS or AS-IV, then co-cultured with primary PMCs in transwell plates. The macrophage-derived exosomes were extracted and purified by differential centrifugation, then co-cultured with primary PMCs. Small RNA-seq was used to detect differential miRNAs in exosomes, and then KEGG analysis and q-PCR were performed for validation. In vivo PD rat models were established by inducing with high-glucose peritoneal dialysis fluid and different concentrations of AS-IV and exosomes were intraperitoneal injection. Through qRT-PCR, western blotting, and luciferase reporting, candidate proteins and pathways were validated in vivo and in vitro. The functions of the validated pathways were further investigated using the mimic or inhibition strategy. PF and inflammatory situations were assessed.
We found AS-IV reversed the MMT of PMCs caused by LPS-stimulated macrophages and the improving effect was mediated by macrophage-derived exosomes in vitro. We also demonstrated that AS-IV significantly reduced the MMT of PMCs in vitro or PF in a rat PD model via regulating exosome-contained miR-204-5p which targets Foxc1/β-catenin signaling pathway.
AS-IV attenuates macrophage-derived exosomes induced fibrosis in PD through the miR-204-5p/Foxc1 pathway.
腹膜透析（PD）是终末期肾病的一种成功的肾脏替代疗法。长期 PD 会导致腹膜间皮细胞（PMCs）的上皮-间充质转化（MMT），导致腹膜纤维化（PF），从而降低 PD 的效率。巨噬细胞被认为在腹膜损伤的发生和持续中起作用。然而，巨噬细胞-PMC 通讯调节腹膜纤维化的机制尚不完全清楚，导致缺乏疾病修饰药物。黄芪甲苷 IV（AS-IV）对 PD 中的 PF 具有抗纤维化作用，而 AS-IV 在 PD 中的作用机制尚不清楚。
从原代巨噬细胞中提取并用 LPS 或 AS-IV 处理，然后在 Transwell 板中与原代 PMCs 共培养。通过差速离心提取和纯化巨噬细胞衍生的外泌体，然后与原代 PMCs 共培养。使用小 RNA-seq 检测外泌体中差异的 miRNAs，然后进行 KEGG 分析和 q-PCR 验证。通过诱导高糖腹膜透析液和不同浓度的 AS-IV 建立 PD 大鼠模型，并进行腹腔内注射外泌体。通过 qRT-PCR、western blotting 和荧光素酶报告，在体内和体外验证候选蛋白和途径。进一步使用模拟物或抑制策略研究验证途径的功能。评估 PF 和炎症情况。
我们发现 AS-IV 逆转了 LPS 刺激的巨噬细胞引起的 PMCs 的 MMT，并且这种改善作用是通过体外巨噬细胞衍生的外泌体介导的。我们还证明，AS-IV 通过调节外泌体中包含的 miR-204-5p 靶向 Foxc1/β-catenin 信号通路，显著降低了体外 PMCs 的 MMT 或 PD 大鼠模型中的 PF。
AS-IV 通过 miR-204-5p/Foxc1 通路减轻巨噬细胞衍生的外泌体诱导的 PD 纤维化。
