UHPLC/Q-TOF-MS-based blood-component profiling and multi-omics analysis reveal potential protective mechanisms of Shenzhuo Formula against diabetic kidney disease.
Shenzhuo Formula shows potential in modulating pathways involved in diabetic kidney disease, but human trials are needed to confirm these findings.
Where it sits
this study against the rest of the vip (vasoactive intestinal polypeptide) corpusSummary and findings
The study investigated the protective mechanisms of Shenzhuo Formula (SZF) against diabetic kidney disease (DKD) using a multi-omics approach. In db/db mice, SZF reduced urinary albumin-to-creatinine ratio and renal injury biomarkers, and improved renal histopathology. Zebrafish models showed improved microcirculatory phenotypes with SZF treatment.
Abstract
Diabetic kidney disease (DKD) is a common microvascular complication of diabetes, with no definitive treatment currently available for DKD-related kidney injury. Podocyte injury, apoptosis, and microcirculation dysfunction play critical roles in the pathogenesis and progression of DKD. Shenzhuo Formula (SZF), a traditional Chinese herbal formulation, has shown clinical potential in reducing proteinuria, but its chemical basis and protective mechanisms remain unclear. This study combined Ultra-high-performance liquid chromatography-high resolution mass spectrometry (UHPLC/Q-TOF-MS) analysis, network pharmacology, renal proteomics, metabolomics, single-nucleus RNA sequencing (snRNA-seq), and in vivo validation to explore the potential mechanisms of SZF against DKD-related renal injury. UHPLC/Q-TOF-MS analysis annotated 27 SZF-related serum constituents, including 19 putative prototype constituents and 8 putative metabolites. Network pharmacology predicted 87 overlapping targets between SZF-related constituents and DKD, mainly enriched in apoptosis, inflammatory responses, focal adhesion, and metabolic regulation. In db/db mice, SZF reduced urinary albumin-to-creatinine ratio and renal injury biomarkers, including NGAL, CysC, and KIM-1, increased estimated glomerular filtration rate, and alleviated glomerular basement membrane thickening, mesangial matrix expansion, inflammatory infiltration, and collagen deposition. Proteomic and metabolomic analyses suggested that SZF regulated pathways related to necroptosis, complement and coagulation cascades, platelet activation, amino acid metabolism, and the citrate cycle. snRNA-seq and histological validation indicated that SZF reduced apoptosis-related gene signatures and podocyte injury. Western blot analysis showed increased BCL-2 expression and decreased cleaved Caspase-3 and cleaved PARP1 expression after SZF treatment. In zebrafish models, SZF improved microcirculatory phenotypes by increasing vessel diameter, cardiac output, and blood-flow velocity, and by reducing thrombus incidence. These hypothesis-generating findings suggest that SZF warrants further investigation as a potential modulator of podocyte apoptosis and microcirculatory dysfunction in DKD.
Background
Diabetic kidney disease (DKD) is a serious complication of diabetes with limited treatment options. Podocyte injury and microcirculation dysfunction are key factors in its progression. Shenzhuo Formula (SZF), a traditional Chinese herbal medicine, has shown potential in reducing proteinuria, but its mechanisms of action are not well understood. This study aims to elucidate the protective mechanisms of SZF against DKD using a comprehensive multi-omics approach.
Methods
The study employed a combination of ultra-high-performance liquid chromatography-high resolution mass spectrometry (UHPLC/Q-TOF-MS), network pharmacology, renal proteomics, metabolomics, and single-nucleus RNA sequencing (snRNA-seq) to investigate SZF's effects. In vivo validation was conducted in db/db mice and zebrafish models. Primary outcomes included changes in urinary albumin-to-creatinine ratio, renal injury biomarkers, and renal histopathology.
Results
SZF treatment in db/db mice resulted in a reduction in urinary albumin-to-creatinine ratio and renal injury biomarkers such as NGAL, CysC, and KIM-1. Histological analysis showed alleviated glomerular basement membrane thickening and mesangial matrix expansion. Proteomic and metabolomic analyses indicated regulation of pathways related to necroptosis and metabolic processes. In zebrafish models, SZF improved microcirculatory phenotypes, including increased vessel diameter and blood-flow velocity.
Interpretation
The findings suggest that SZF may modulate pathways involved in podocyte apoptosis and microcirculatory dysfunction, which are critical in DKD progression. While the results are promising, they are based on animal models, and the clinical relevance in humans remains to be established. Further research, including human trials, is needed to confirm these potential therapeutic effects.
Key findings
- 27 SZF-related serum constituents annotated by UHPLC/Q-TOF-MS.
- 87 overlapping targets predicted between SZF constituents and DKD.
- Reduced urinary albumin-to-creatinine ratio in db/db mice.
- Increased estimated glomerular filtration rate in db/db mice.
- Improved microcirculatory phenotypes in zebrafish models.
Limitations
- Animal model-based study.
- Exploratory findings.
- No human clinical data.
- Potential species-specific effects.
- Hypothesis-generating nature.