Extracellular vesicles (EVs) facilitate intercellular communication by traversing the extracellular matrix (ECM). However, their motility within fibrotic ECM and its role in fibrosis development remain unclear. We engineered stress-relaxing (SR) hydrogels of tunable stiffness (2, 50 kPa) through dynamic crosslinking of short peptide (WGG(KA)) and heparin to mimic normal and fibrotic ECM. Super-resolution nanoimaging and quantitative three dimensional (3D) single-particle tracking (SPT) of single EV were performed, and the motion dynamics was quantified. The interplay between EVs and ECM was further investigated, particularly its effects on fibroblast activation and renal fibrosis. It was identified that both normal and fibrotic kidney-derived EVs exhibited confined Brownian-like motion according to 3D SPT, with enhanced mobility in the stiffer (50 kPa) hydrogel. Notably, fibrotic tubule-derived EVs carried higher levels of integrin β6 (ITGB6), which reduced their mobility within the hydrogel-based ECM mimic, as confirmed by the restoration of motility upon ITGB6 blocking or digestion. This suggested that EV motility may be influenced by the interplay between ECM stiffness and the intrinsic properties of the EVs. Furthermore, enrichment of ITGB6 on fibrotic tubule-derived EVs promotes local retention, thereby increasing EV-fibroblast interaction and profibrotic signaling. This indicated the underappreciated role of EV in fibrosis related to its motility and its interplay with fibroblast in fibrotic niche. Our study provides new insights into the mechano-dependent mechanisms governing EV motility within the fibrotic ECM. STATEMENT OF SIGNIFICANCE: Extracellular vesicles (EVs) play key roles in cell communication, but how they move through fibrotic tissue remains poorly understood. This study reveals that kidney-derived EVs exhibit confined Brownian-like motion within engineered hydrogels mimicking fibrotic extracellular matrix. We found that EVs from fibrotic tubules carry elevated integrin β6, which restricts their mobility and promotes pro-fibrotic signaling by increasing EV-fibroblast interaction. Our study provides new insights into the mechano-dependent mechanisms governing EV motility within the fibrotic ECM. This work provide a biophysical perspective for understanding EVs mediated pathological communication in fibrosis.

Sepsis-induced acute kidney injury (S-AKI) is a life-threatening condition driven by excessive immune inflammation, and effective treatments remain lacking. Mesenchymal stem cell-derived small extracellular vesicles (MSC-sEV) have been demonstrated to possess potent immunomodulatory activity. This study aimed to investigate the role and underlying mechanism of MSC-sEV in S-AKI. We established in vivo and in vitro models of S-AKI and employed techniques such as small RNA sequencing, transcriptome sequencing, luciferase reporter assays, and engineered gene editing to validate therapeutic efficacy and elucidate mechanisms. Results demonstrated that in S-AKI, MSC-sEV homed to injured kidneys and were internalized by renal tubular epithelial cells, significantly ameliorating renal damage and improving survival rates. Mechanistically, MSC-sEV delivered miR-125a-5p to target and inhibit TNFR2 expression, thereby blocking TNF-driven pyroptosis mediated by the NF-κB/NLRP3 signaling pathway. Furthermore, engineered modification with the EXOMotif GGAG significantly enhanced MSC-sEV delivery of miR-125a-5p and inhibition of TNFR2. In conclusion, this study demonstrates that MSC-sEV represent a promising drug delivery vehicle with substantial targeted therapeutic potential for S-AKI.

Acute kidney injury (AKI) remains a major clinical challenge due to the lack of effective interventions. While mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) show therapeutic promise for AKI, their exact mechanisms are largely to be understood. Human umbilical cord-derived MSC-EVs were isolated, characterized, and tested in a murine bilateral renal ischemia reperfusion injury (bIRI) model and in hypoxia/reoxygenation (H/R) treated tubular epithelial cells in vitro. Integrated transcriptomic, miRNA, and biochemical analyses were performed to elucidate the metabolic pathways and molecular mechanisms underlying the renoprotective effects of MSC-EVs. MSC-EVs preferentially targeted injured kidneys and significantly improved renal function, ameliorated tubular injury, and suppressed inflammation in IRI-AKI. RNA sequencing and targeted metabolomics revealed substantial dysregulation of steroid metabolism after IRI, marked by activation of the cholesterol 25-hydroxylase (CH25H)/25-hydroxycholesterol (25HC) axis. Importantly, accumulated 25HC induced lipid peroxidation and ferroptosis in tubular epithelial cells. MSC-EVs treatment reversed these pathological changes by downregulating CH25H, lowering 25HC levels, and restoring redox homeostasis. miRNA profiling further identified miR-26b-5p as a key MSC-EVs cargo that directly targets the 3′UTR of CH25H mRNA to repress its expression. Notably, inhibiting miR-26b-5p within EVs abrogated their ability to suppress CH25H/25HC-driven ferroptosis, thereby demonstrating its essential role in the metabolic and cytoprotective actions of MSC-EVs. Our findings unveil the CH25H/25HC axis as a key metabolic checkpoint governing tubular ferroptosis in ischemic AKI. MSC-EVs deliver miR-26b-5p to suppress this axis, thereby rectifying oxysterol metabolism and preventing ferroptosis.

The plasma protein corona (PC) critically influences the in vivo fate of nanomedicines, yet its composition and impact on extracellular vesicles (EVs) remain poorly defined. Using a biomimetic circulation system, we characterized PC formation and modulation on two clinically relevant EV types: mesenchymal stromal cell-derived EVs (MSC-EVs) and HEK293F-derived EVs (293F-EVs). Under dynamic flow, both EV types acquired stable coronas, resulting in increased particle size and decreased surface charge. Proteomic profiling revealed a shared corona signature enriched in immunoglobulins, complements, and other plasma components. Functionally, PC formation enhanced macrophage uptake and triggered inflammatory activation, primarily via interactions between corona-bound immunoglobulins or complement C3 and their respective receptors. To disrupt this process, we developed a charge-shielding strategy using positively charged chitosan oligosaccharide (COS) to inhibit PC assembly. COS coating effectively neutralized EV surface charge and reduced opsonin adsorption and non-specific macrophage clearance, thereby reshaping EV biodistribution-limiting hepatic sequestration and enhancing delivery to extrahepatic organs. In a murine sepsis model, COS-modified MSC-EVs further improved renal and pulmonary outcomes and markedly increased survival. Collectively, these findings elucidate the molecular architecture and immunological impact of the EV-associated plasma PC and introduce a promising anti-corona strategy for engineering stealthier and more effective EV-based nanotherapeutics.

BACKGROUND:Renal fibrosis (RF) is a progressive pathological process driven by chronic inflammation and Th17/Treg imbalance. Asiaticoside (AS), a triterpenoid compound from Centella asiatica (L.) Urb., exhibits anti-inflammatory and antifibrotic activities, though its molecular mechanism remains unclear. OBJECTIVE:This study aimed to investigate whether AS alleviates RF by targeting Signal transducer and activator of transcription 3 (STAT3) through a "bind to destabilize" mechanism to restore Th17/Treg homeostasis. METHODS:An integrated approach combining network pharmacology, transcriptomics, and multimodal experimental validation was applied. UUO mice were treated with AS (10, 50, 100mg/kg/d) for 10 days. Histopathology, RNA‑seq, flow cytometry, immunofluorescence, Luminex, qPCR, DARTS‑LC‑MS/MS, molecular docking/dynamics simulations and SPR were performed. Pharmacological interventions using Stattic (STAT3 inhibitor) and Colivelin (STAT3 agonist) were included to functionally validate the role of STAT3. RESULTS:Network pharmacology identified STAT3 as the core target, with Th17 differentiation as the key pathway. AS treatment significantly attenuated RF, improved renal function, and rebalanced Th17/Treg ratios in UUO mice, accompanied by reduced IL-17A and elevated IL-10. Transcriptomic analysis revealed enriched Th17 cell differentiation genes, validated by qPCR. DARTS-LC-MS/MS confirmed direct binding of AS to STAT3 and identified a peptide derived from the SH2 domain (residues 582-602), indicating conformational destabilization. SPR showed high affinity binding to both human and murine STAT3. Molecular docking and dynamics simulations demonstrated a "local anchoring-allosteric effect" mode within the SH2 domain. qPCR analysis showed that AS significantly inhibited the mRNA expression of both IL-17A and total STAT3 in the renal tissues of UUO mice. Immunofluorescence revealed reduced STAT3 and p-STAT3 expression in kidneys. STAT3 inhibitor Stattic mimicked AS's antifibrotic and Th17 suppressive effects, whereas agonist Colivelin exacerbated fibrosis and was partially rescued by AS. CONCLUSION:AS alleviates RF via a novel "bind to destabilize" allosteric degradation mechanism that directly targets the STAT3 SH2 domain. This interaction induces conformational instability, suppresses STAT3 activation and transcriptional activity, restores Th17/Treg homeostasis, and ultimately mitigates renal inflammation and fibrosis. Collectively, these findings establish a new therapeutic strategy for STAT3-driven fibrotic diseases.

IntroductionVascular calcification (VC) is a prevalent and life-threatening complication of chronic kidney disease (CKD), yet the mechanisms by which hyperphosphatemia drives VC remain incompletely understood. This study investigates the role of endothelial cells (ECs)-derived exosomal microRNAs in mediating osteogenic differentiation of vascular smooth muscle cells (VSMCs) under high phosphate (HP) conditions.MethodsA CKD-VC mouse model was established using a HP and high-adenine diet. Exosomes (Exos) were isolated from ECs cultured under normal or HP conditions. The effects of Exos on calcification of VSMCs were evaluated using in vitro co-culture systems and in vivo administration. miRNA sequencing, dual-luciferase reporter assays, and loss/gain of function experiments were performed to identify key exosomal miRNAs and their downstream targets. Western blotting, qRT-PCR, and histological analyses were used to assess molecular and pathological changes.ResultsHP-stimulated ECs released Exos (HP-Exos) that were internalized by VSMCs and significantly promoted VC in both in vitro and in vivo models. miRNA sequencing identified miR-299-3p as significantly upregulated in HP-Exos. Functional studies demonstrated that exosomal miR-299-3p directly targeted membrane-associated RING-CH3 (MARCH3), leading to activation of the p-JAK2/STAT5 signaling pathway. This cascade subsequently upregulated osteogenic markers and downregulated contractile marker, thereby promoting osteogenic differentiation of VSMCs. Knockdown of miR-299-3p in vivo attenuated VC in CKD mice.DiscussionThese findings reveal a previously unrecognized mechanism by which HP drives CKD-VC through ECs-derived exosomal miR-299-3p. The miR-299-3p/MARCH3/p-JAK2/STAT5 signaling axis represents a critical regulatory pathway in VC pathogenesis and offers a potential therapeutic target for this life-threatening complication of CKD.

Acute kidney injury (AKI) occurs in the patients undergoing anti-programmed cell death protein 1-ligand 1 (PD-L1) therapy, indicating that PD-L1 may play a critical role in maintaining renal homeostasis. However, the precise role and mechanism of PD-L1 in AKI remains largely elusive. In this study, we found that PD-L1 was primarily expressed in proximal tubules and significantly upregulated in both murine models of AKI and renal biopsy samples from AKI patients. Genetic specific deletion of PD-L1 in mouse tubular epithelial cells (TECs) exacerbated renal injury in ischemia-reperfusion injury-induced AKI. Mechanistically, PD-L1 was found to interact with BRCA1 and increase BRCA1 expression to safeguard TECs against DNA damage, thereby promoting cellular proliferation and suppressing apoptosis. To translate these findings into a potential therapeutic strategy, we developed a CGA-functionalized extracellular vesicle delivery system for targeted delivery of PD-L1 to injured TECs. This system efficiently restored PD-L1 expression and alleviated DNA damage of TECs in both TEC-specific PD-L1 knockdown and T-cell knockout AKI mouse models. Collectively, these findings uncover a novel function of PD-L1 in promoting adaptive TEC repair through BRCA1 interaction, independent of its canonical immunomodulatory function of T cells, and suggest that PD-L1 supplementation may represent a promising therapeutic strategy for AKI.

The increasing global impact of kidney diseases highlights a pressing and unmet need for new treatment strategies. In this context, mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have emerged as a promising cell-free therapeutic approach, attracting growing interest due to their dual regenerative functions. MSC-EVs not only possess intrinsic therapeutic effects mediated by their cargo of mRNAs, proteins, and miRNAs that regulate inflammation, promote cell repair, reduce fibrosis, but also serve as highly biocompatible vehicles for drug delivery. This versatility places them at the forefront of a potential shift in how kidney diseases may be treated. In this review, we systematically summarize current knowledge on the mechanisms and therapeutic potential of MSC-EVs in various kidney diseases. Although challenges related to standardization and clinical translation persist, ongoing progress supports the view that MSC-EVs are poised to become key next-generation therapies for kidney diseases.

ObjectiveTo evaluate the synergistic immunological protection of exosomal T-cell epitope vaccine and antibody-inducing vaccine against SARS-CoV-2 in highly humanized mice.MethodsRed blood cell-derived exosomes were loaded with 27 CD8+ T-cell epitope peptides and 19 CD4+ T-cell epitope peptides followed by combined immunization with S1 protein vaccine of SARS-CoV-2 and adjuvant poly I:C in HLA-A2/DR1 double transgenic mice. After immunizations, splenocytes were assessed for epitope-specific T cell responses by intracellular cytokine staining and ELISPOT, and for functional T cell subset analysis through flow cytometry. Meanwhile, serum anti-S1 protein IgG and neutralizing antibodies were quantified via ELISA and BA.5 pseudovirus neutralization assay, respectively. Furthermore, viral challenge was performed after the combined immunization in HLA-A2/DR1/hACE2 triple transgenic mice, followed by viral load quantification, viral protein detection, and H&E staining in lungs.ResultsThe combined immunization i) increased the titers of S1 protein-specific IgG antibodies and neutralizing antibodies as well as Tfh cell frequency as compared to the S1 protein vaccine alone; ii) induced significantly more S1 protein-specific T cells and effector memory CD4+ T cells, and inhibited T cell exhaustion and regulatory T cell differentiation, compared to the exosomal T-cell epitope vaccine alone; iii) achieved the lowest pulmonary viral loads, inflammatory cell infiltration, and histopathological damage after SARS-CoV-2 infection.ConclusionThis study, for the first time, demonstrates the synergistic humoral and cellular immune responses and protective efficacy induced by the combined immunization of exosomal T-cell epitope vaccine and antibody-inducing vaccine, and provides preclinical evidence from highly humanized mice for optimizing next-generation SARS-CoV-2 vaccine protocols.

Renal aging involves structural and functional kidney decline (reduced size/nephrons, glomerulosclerosis, tubular atrophy, functional loss) and is an independent risk factor for kidney and systemic degenerative diseases. On 16 May 2025, the Aging Biomarker Consortium convened an expert consensus meeting in Shanghai, proposing a multidimensional biomarker framework: functional (estimated glomerular filtration rate, renal blood flow), structural (renal volume loss), and humoral (Klotho, N-terminal Pro-B-type natriuretic peptide, senescence-associated secretory phenotype factors). The consensus also supports machine-learning models for biological age assessment and calls for multi-center cohorts and translational collaboration to improve elderly kidney health.

Plasma membrane repair is crucial for resealing membrane disruptions from physiological and pathological stimuli to preserve cell integrity and homeostasis. Tubular epithelial cells (TECs) die of unrepaired membrane injury induced by biochemical and immune factors, leading to the onset and progression of acute kidney injury (AKI). Indeed, mammalian cells are equipped with repair pathways and molecular machinery to safeguard cell viability. Depending on the severity and nature of plasma membrane injury, membrane disruptions can be resealed by vesicle-dependent and independent approaches. Besides, the process of membrane resealing is also important for the repair of damaged organelle membranes. Herein, different formats of plasma membrane damage were discussed, highlighting the membrane disruption induced by pore-forming proteins (PFPs), including MAC, perforin, and membrane-damaging proteins in regulated cell death (RCD). Moreover, the mechanisms of plasma and organelle membrane repair to guard against the death of TECs in AKI were discussed, with the aim of proposing novel strategies for AKI therapy.

BACKGROUND:The activation of the renin-angiotensin system (RAS) and lipid disorders are major risk factors in progressive chronic kidney disease. This study aimed to investigate the potential synergistic mechanisms of RAS activation and lipid disorders that contribute to glomerulosclerosis. MATERIALS AND METHODS:Human renal mesangial cells (HMCs) were treated with 10(-7) mol/L angiotensin II (Ang II) or with 30 μg/ml cholesterol and 1 μg/ml 25-hydroxycholesterol (lipid loading) for 24 hours. Lipid accumulation in the cells was evaluated by Oil Red O staining and intracellular cholesterol quantitative assays. The gene and protein expression of molecules in the low-density lipoprotein receptor (LDLr) pathway, the RAS family, and the extracellular matrix were examined by real-time polymerase chain reaction and Western blotting. The translocation of sterol regulatory element-binding protein (SREBP) cleavage activating protein (SCAP), which escorts SREBP-2 from the endoplasmic reticulum (ER) to the Golgi, was examined by immunofluorescent staining. RESULTS:Ang II increased lipid droplet accumulation in HMCs. Further analysis revealed that Ang II increased the mRNA and protein expression of LDLr, SCAP, and SREBP-2. This increase was correlated with an enhanced translocation of the SCAP/SREBP-2 complex from the ER to the Golgi in HMCs that was induced by Ang II, thereby activating LDLr gene transcription. Interestingly, lipid loading increased the mRNA and protein expression of angiotensinogen, Ang II, renin, angiotensin-converting enzyme, angiotensin II type 1 receptor, and type 2 receptor in HMCs with increased mRNA and protein expression of collagen I, α-smooth muscle actin, and fibronectin. CONCLUSIONS:This study demonstrates that the interaction of RAS activation and lipid disorders accelerates the progression of glomerulosclerosis.

Key PointsIntegrated single-cell spatial transcriptomics and metabolomics identified cholesterol reprogramming as a driver of ferroptosis in proximal tubules.Targeting the Cyp7b1/27-hydroxycholesterol axis efficiently alleviated ferroptosis-induced proximal tubule injury during ischemic AKI.BackgroundCell death plays a pivotal role in ischemic AKI, with metabolic dysfunction emerging as a key contributor. However, the mechanism by which metabolism imbalance initiates renal tubular cell death is poorly understood.MethodsWe combined single-cell spatial transcriptomics and metabolomics to characterize the function and metabolites of murine renal proximal cell subpopulations during ischemic AKI to CKD transition.ResultsFerroptosis was identified as the predominant mode of cell death in severely injured proximal straight tubules after AKI. Additional investigation revealed a critical deficiency in Cyp7b1, an enzyme responsible for metabolizing 27-hydroxycholesterol (27-HC) into 7 alpha,27-dihydroxycholesterol, resulting in substantial 27-HC accumulation in proximal tubular cells during the early phase of ischemic AKI. Mechanistically, 27-HC acts as an endogenous ligand for estrogen receptor alpha, inducing downstream Hmox1 activation and thereby potentiating ferroptosis susceptibility in proximal tubular cells. Notably, adeno-associated virus-mediated Cyp7b1 overexpression in a murine ischemia-reperfusion injury model attenuated ferroptosis by enhancing 27-HC degradation, effectively mitigating ischemic AKI progression. These findings underscore the pivotal role of the Cyp7b1/27-HC axis in this pathologic context.ConclusionsOur study delineated a unique mechanism of Cyp7b1/27-HC axis in proximal tubular cell ferroptosis in early AKI.

Acute kidney injury (AKI) is a severe clinical event with a high risk of transitioning to chronic kidney disease (CKD). Persistent hypoxia resulting from peritubular capillaries (PTCs) loss is a key factor driving this chronic transition. However, the mechanisms governing PTCs injury and repair, particularly under different AKI severity, remain unclear. This study demonstrates that small extracellular vesicle (sEV) carried VEGF-A as an essential regulator of PTCs repair and decreased as the severity of tubular injury increased. We found that VEGF-A was sorted into sEV via syndecan-1 (SDC-1), attaching to the heparan sulfate chain in moderate AKI. SDC-1 knockdown in renal tubules significantly reduced VEGF-A packaging into EVs and aggravated PTCs rarefaction and renal fibrosis. Overexpression of SDC-1 promoted the production of VEGF-A+ sEV and their proliferative effects on endothelial cells. These findings demonstrate that SDC-1 contributes to the secretion of VEGF-A+ sEV from moderately injured tubules, facilitating PTCs repair after AKI.

Mesenchymal stem cell-derived extracellular vesicles (MSC-EVs) have emerged as a promising approach in regenerative therapy. However, the clinical application of MSC-EVs is hindered by the presence of xenogenic components, such as fetal bovine serum (FBS), which is the most used culture supplement for MSCs. Human platelet lysate (HPL) has been proposed as an alternative to FBS, but whether MSC-EVs derived from HPL-cultured MSCs are suitable for clinical translation remains unclear. In this study, we comprehensively compared the characterization of EVs derived from MSCs cultured in the medium with FBS (F-EVs) and HPL (H-EVs). Our study showed that HPL promoted MSC-EV production without compromising EVs critical quality attributes. Multiomics sequencing revealed the stability of H-EVs from different umbilical cord donors and global functional alterations for MSC-EVs under different culture conditions. In comparison to F-EVs, H-EVs enriched more angiogenesis-related molecules and exhibited enhanced angiogenesis, which were further confirmed by in vivo and in vitro studies. H-EVs significantly reduced renal microvascular rarefaction and promoted the regeneration of umbilical vein endothelial cells to hypoxia stimulation compared to that of F-EVs. In conclusion, our findings demonstrated that HPL as culture supplements did not alter the critical quality attributes of MSC-EVs, specifically holding a higher yield and quality of MSC-EVs with enhanced angiogenic potential.

Sepsis-associated acute kidney injury (SA-AKI) portends severe health burden due to significant morbidity and mortality, while early diagnosis remains challenging. In this study, proximity-dependent barcoding assay (PBA) is established to profile the surface proteome of single urinary extracellular vesicle (uEV). Principle uEV clusters with unique function and origination are profiled in SA-AKI in a screening cohort. Complement receptor CD35 on single uEV (CD35-uEV) displays high diagnostic accuracy for SA-AKI (AUC-ROC 0.89 in validation cohort, n = 134). Besides, CD35-uEV enables identification of subclinical AKI (AUC-ROC 0.84 in prospective cohort, n = 72). Moreover, CD35-uEV correlates closely with AKI severity which also predicts persistent AKI (AUC-ROC 0.77), mortality risks (AUC-ROC 0.70) and progression to AKD (AUC-ROC 0.66). Multi-omics profiling reveals that CD35-uEV are predominantly released from injured podocytes exhibiting diminished CD35 expression. Overall, this study identifies a single uEV biomarker related to injured podocyte for early diagnosis and risk stratification of SA-AKI.

Macrophages are heterogeneous and play pivotal roles in renal injury and repair, making them prime therapeutic targets for kidney disease. However, their diversity and function in the transition from acute kidney injury (AKI) to chronic kidney disease (CKD) remain poorly understood. Here, we employed single-cell RNA-sequencing (scRNA-seq) to identify key macrophage subpopulations involved in renal repair after AKI. We also explored a chemical approach to reprogram macrophages for AKI recovery. A mouse model of renal ischemia-reperfusion AKI (IR-AKI) was established. Renal tissue was collected at different time points post-AKI for scRNA-seq analysis. Macrophages were identified and clustered based on gene expression profiles. Gene enrichment analysis and Cellchat were performed for the identification of pro-repair macrophage. Key transcription factors (TFs) were predicted to understand the gene regulatory network of pro-repair macrophages. To induce a pro-repair macrophage phenotype, small molecule compounds targeting key TFs and repair-associated factors were applied to RAW cells in various combinations. The efficacy of these chemical cocktails was evaluated by repair factors levels and transcriptome sequencing analysis. The repair function of chemically reprogrammed macrophages was validated in vitro using co-culture systems with tubular epithelial cells or vascular endothelial cells. Finally, chemical cocktails were administered to AKI mice to reprogram macrophages into a pro-repair functional phenotype to facilitate renal repair. We identified 2241 macrophages through renal scRNA-seq, which clustered into seven distinct subgroups. A pro-repair macrophage subpopulation with unique renal repair functions was defined, characterized by high expression of epithelial repair factors (Hbegf, Gdf15, Spink1) and angiogenic factors (Vegfa, Ccn1, Timp3, Adamts1). Cellchat analysis showed that pro-repair macrophages have strong interactions with damaged tubular epithelial cells and endothelial cells. Gene regulatory network analysis identified Cebpb, Ppara and Egr1 as core transcription factors governing pro-repair macrophage function. Furthermore, we screened fifteen combinations of small molecule drugs and found that the FDRC chemical cocktail effectively activates Cebpb, Ppara and EGR1 in RAW cells, leading to significant expression of Hbegf, Gdf15, and Vegfa. RNA-seq found that FDRC-reprogrammed macrophages display a unique gene expression profile distinct from M0 or M2 macrophages, which significantly increased gene expressions related to angiogenesis, anti-inflammatory, and wound healing. FDRC-reprogrammed RAW cells can significantly promote their proliferation when co-cultured with endothelial cells and tubular epithelial cells. Notably, FDRC successfully reprogrammed macrophages toward a pro-repair-like phenotype in IR-AKI mice, which significantly ameliorated tubular injury and peritubular capillary rarefaction, and potently prevented the AKI-to-CKD transition. Our study uncovers a novel pro-repair macrophage subset during AKI and introduces a promising chemical reprogramming cocktail to manipulate macrophage phenotype in vivo, facilitating the repair of renal tubules and peritubular capillaries.