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

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 are 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 interlukin-1 beta (IL-1 beta) in renal macrophage polarization during diabetic nephropathy. B4 effectively mitigated macrophage polarization, alleviating inflammation and fibrosis, confirmed by Western blot. Fibroblast analysis demonstrated reduced fibrosis with B4 treatment, consistent with hematoxylin and eosin staining and Immunofluorescence 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. In the current study, the author combined single-cell RNA-sequencing (scRNA-seq) with proteomics technic to decipher the cellular events during diabetic progression with Anemoside B4 treatment. Their findings provide a comprehensive insight into treating diabetic nephropathy with Anemoside B4 via alleviating inflammation and fibrosis by mitigating macrophage polarization and addressing oxidative stress.image