Department of Nephrology, Beijing Friendship Hospital,Capital Medical University, No. 95, Yong'an Road, Xicheng District, 100050, Beijing, PR China.
BMC Nephrol. 2023 Jan 30;24(1):24. doi: 10.1186/s12882-023-03065-w.
Macrophages contribute to epithelial-mesenchymal transition (EMT) in diabetic nephropathy (DN). Exosomal long non-coding RNAs (lncRNAs) derived from macrophages play a major role in transmitting biological information, whereas related studies on DN are rare. Here we investigated the effects of exosomal lncRNAs from high glucose-treated macrophages on EMT.
High glucose-treated macrophage exosomes (HG-exos) were extracted by coprecipitation and stabilized. Then, mouse renal tubular epithelial cells were treated with HG-exos for 24 h. Expression of E-cadherin, α-smooth muscle actin (α-SMA), and fibronectin was detected by western blotting, qPCR, and immunofluorescence. High-throughput sequencing was then applied to analyze the bioinformatics of HG-exos.
HG-exos inhibited the proliferation of tubular epithelial cells. Additionally, HG-exos markedly upregulated α-SMA and fibronectin expression and downregulated E-cadherin expression in tubular epithelial cells, indicating EMT induction. A total of 378 differentially expressed lncRNAs and 674 differentially expressed mRNAs were identified by high-throughput sequencing of HG-exos. Bioinformatics analysis and subsequent qPCR validation suggested 27 lncRNAs were enriched in the EMT-related MAPK pathway. Among them, ENSMUST00000181751.1, XR_001778608.1, and XR_880236.2 showed high homology with humans.
Exosomes from macrophages induce EMT in DN and lncRNAs in exosomes enriched in the MAPK signaling pathway may be the possible mechanism.
巨噬细胞参与糖尿病肾病(DN)中的上皮间质转化(EMT)。巨噬细胞来源的外泌体长链非编码 RNA(lncRNA)在传递生物学信息方面发挥着重要作用,而关于 DN 的相关研究却很少。本研究旨在探讨高糖处理的巨噬细胞来源的外泌体 lncRNA 对 EMT 的影响。
采用共沉淀法提取并稳定高糖处理的巨噬细胞外泌体(HG-exos)。然后,用 HG-exos 处理小鼠肾小管上皮细胞 24 h。Western blot、qPCR 和免疫荧光检测 E-钙黏蛋白、α-平滑肌肌动蛋白(α-SMA)和纤连蛋白的表达。随后,采用高通量测序分析 HG-exos 的生物信息学。
HG-exos 抑制肾小管上皮细胞的增殖。此外,HG-exos 明显上调了肾小管上皮细胞中 α-SMA 和纤连蛋白的表达,下调了 E-钙黏蛋白的表达,表明 EMT 的诱导。通过对 HG-exos 的高通量测序,共鉴定出 378 个差异表达的 lncRNA 和 674 个差异表达的 mRNA。生物信息学分析和随后的 qPCR 验证表明,27 个 lncRNA 富集在 EMT 相关的 MAPK 通路中。其中,ENSMUST00000181751.1、XR_001778608.1 和 XR_880236.2 与人具有高度同源性。
巨噬细胞来源的外泌体在 DN 中诱导 EMT,外泌体中的 lncRNA 可能富集在 MAPK 信号通路中,这可能是其作用机制。