Research on the Progression of Renal Fibrosis Disease Based on Metabolomics and Transcriptomics and the Dynamic Intervention of Amygdalin

Zhenyu LeiJia WangHongbing ZhouJiaxing GaoRunxi WangShuyuan JiangWanfu BaiYingchun BaiLiya FanChang HongSongli Shi^*

• Baotou Medical College, Baotou, China

The primary objective of this study is to examine the alterations in differential metabolites and differentially expressed genes at various stages of renal fibrosis (RF) in unilateral ureteral obstruction (UUO) rat models and the effects of amygdalin treatment.A total of forty-eight male Sprague-Dawley (SD) rats were randomly allocated into six groups based on body weight: sham surgery group (SDG), model group (MOD), benazepril hydrochloride group (BHT), low-dose amygdalin group (AMY-L), medium-dose (AMY-M), and highdose (AMY-H), with eight rats in each group. All groups, except the sham surgery group, underwent Abbreviations: RF, Renal fibrosis; AMY, Amygdalin; UUO, unilateral ureteral obstruction; CKD, Chronic kidney disease; SD, Sprague-Dawley; ADG, surgery group; MOD, model group; AMY-L, low-dose AMY group; AMY-M, medium-dose amygdalin group; AMY-H, high-dose amygdalin group; BHT, benazepril hydrochloride group; AMY1, amygdalin medium dose for one week; AMY2, amygdalin medium dose for two weeks; AMY4, amygdalin medium dose for four weeks; UPLC-Q-TOF-MS, ultra-performance liquid chromatography/quadrupole time-of-flight-tandem mass spectrometry; VIP, variable importance analysis; PCA, principal component analysis; KEGG, Kyoto encyclopedia of genes and genomes; OPLS-DA, partial least squares discriminant analysis; DEGs, differentially expressed genes. left ureteral obstruction to establish the model of RF. The kidney and blood samples of the rats were taken in batches for one week, two weeks, and four weeks after continuous intragastric administration.Pharmacodynamic assessment, metabolomics, and transcriptomic analysis were performed after sample treatment. Western Blot detects the expression of PPARγ, PGC-1α, TGF-β1, and p-Smad3related proteins in kidney tissue.As RF progresses, inflammatory and fibrotic factors exhibit stage-dependent elevation. AMY demonstrates therapeutic potential by attenuating renal interstitial/tubular damage and fibrosis severity. Integrated metabolomic-transcriptomic analyses revealed RF pathophysiology is associated with dysregulation of sphingolipid, linoleic acid, and glutathione metabolism pathways.AMY primarily exerts anti-fibrotic effects through sphingolipid metabolism modulation, evidenced by increased sphingosine 1-phosphate levels, altered dihydroceramide profiles, and upregulation of Cers3/Sphk1 expression. Mechanistically, AMY treatment upregulated PPARγ and PGC-1α protein expression while suppressing TGF-β1/p-Smad3 signaling in renal tissues. These findings position linoleic acid metabolism as a critical pathway in RF progression and establish AMY's efficacy through sphingolipid-mediated metabolic reprogramming.