Single‐Cell Transcriptomic Analysis Reveals the Alleviating Effect of Anemoside B4 on Murine Diabetic Nephropathy

Abstract

In Diabetic Kidney Disease (DKD), inflammation exacerbated by oxidative stress (ROS) is pivotal. This study investigates the impact of Anemoside B4 (B4), an anti‐inflammatory drug, on kidney cell populations in DKD mice using single‐cell RNA sequencing (scRNA‐seq) and mass spectrometry‐based proteomics, focusing on its hypoglycemic, anti‐inflammatory, and anti‐fibrotic properties. Compared to the DKD group, B4 treatment reduces blood lipid metabolism levels, renal inflammation, and fibrosis in mice. Through scRNA‐seq, 14 distinct renal cell clusters were identified, primarily proximal tubular (PT) cells. In DKD mice, PT subtype cells exhibited elevated SPP1 expression, promoting macrophage M1 polarization, inflammatory cell infiltration, and renal fibrosis—all reversed by B4. Macrophage analysis revealed enrichment of the marker gene IL‐1b in renal macrophage polarization during diabetic nephropathy. B4 effectively mitigated macrophage polarization, alleviating inflammation and fibrosis, confirmed by Western blot (WB). Fibroblast analysis demonstrated reduced fibrosis with B4 treatment, consistent with Hematoxylin and Eosin (HE) staining and Immunofluorescence (IF) findings. Notably, oxidative stress contributed to these changes. In conclusion, B4 alleviated kidney inflammation and fibrosis in DKD mice by mitigating macrophage polarization and addressing oxidative stress, providing valuable insights into its multifaceted therapeutic potential.This article is protected by copyright. All rights reserved