Determination of urinary peptides in patients with proteinuria

Prakash, Mungli; Shetty, JK; Dash, Sambit; Barik, B K; Sarkar, Abhirup; Prabhu, Ravindra

doi:10.4103/0971-4065.45289

Cited by 8 publications

(7 citation statements)

References 18 publications

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“…Urine albumin was measured using the Bradford assay (Bio-Rad Laboratories, Hercules, California, USA). 20 …”

Section: Methodsmentioning

confidence: 99%

“…Urine albumin was measured using the Bradford assay (Bio-Rad Laboratories, Hercules, California, USA). 20 Assessment of serology by ELISA ANA, antidouble-strand DNA (dsDNA), and IgG in the plasma were detected using ELISA kits according to the manufacturer's instructions (MyBioSource, San Diego, California, USA). In addition, serum levels of mouse IL-6 and monocyte chemoattractant protein-1 (MCP-1) were examined using ELISA kits according to the manufacturer's instruction (Thermo Fisher Scientific Inc, Waltham, Massachusetts, USA).…”

Section: Assessment Of Proteinuriamentioning

confidence: 99%

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Lupus pathogenesis and autoimmunity are exacerbated by high fat diet-induced obesity in MRL/lpr mice

et al. 2023

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ObjectiveSLE is an autoimmune disease characterised by persistent inflammation and autoantibody production. Genetic predisposition and environmental factors such as a high-fat diet (HFD) may contribute to lupus development. However, the immune cell profile and gender difference in response to HFD in lupus have not been reported. Here we investigated the impact of HFD on lupus pathogenesis and autoimmunity using lupus-prone mice.MethodsThirty male and 30 female MRL/lymphoproliferation (lpr) mice were fed with regular diet (RD) or HFD. Body weights were recorded weekly. SLE progression was monitored by skin lesion, urine protein, titres of antidouble-strand DNA (dsDNA) and ANA. At week 14, kidney and skin tissue sections were stained with H&E and periodic acid–Schiff to detect histological kidney index and skin score. Splenocytes were identified by immunofluorescence staining and flow cytometry.ResultsHFD significantly increased body weight and lipid levels compared with RD (p<0.01). Skin lesions were observed in 55.6% of the HFD group compared with 11.1% of the RD group, with greater histopathological skin scores in the female HFD group (p<0.01). Although both male and female mice had higher serum IgG in the HFD group than in the RD group, only the male HFD group showed an increased trend in anti-dsDNA Ab and ANA titres. Kidney pathological changes in the HFD group were more severe in male mice than in female mice (p<0.05), detected by proteinuria, kidney index and glomerular cell proliferation. Significant increases of germinal centre B cells and T follicular helper cells were observed in the spleens of HFD mice (p<0.05).ConclusionHFD induced an accelerated and exacerbated lupus development and autoimmunity in MRL/lpr mice. Our results parallel many known clinical lupus phenotypes and sexual dimorphism in which male patients are likelier to have a severe disease (nephritis) than female lupus patients who may have a broader range of lupus symptoms.

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“…Urine albumin was measured using the Bradford assay (Bio-Rad Laboratories, Hercules, California, USA). 20 …”

Section: Methodsmentioning

confidence: 99%

Section: Assessment Of Proteinuriamentioning

confidence: 99%

Lupus pathogenesis and autoimmunity are exacerbated by high fat diet-induced obesity in MRL/lpr mice

et al. 2023

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“…Microscopic haematuria was defined as >5 red blood cells (RBCs) per high power field (HPF, 20×). Quantitative measurements for proteinuria were performed using the Bradford assay (Bio‐Rad) 8 . Microscopic examination of urine, on a phase contrast microscope, was performed on urine samples that had been centrifuged for 5 minutes at 2000 rpm.…”

Section: Methodsmentioning

confidence: 99%

“…Quantitative measurements for proteinuria were performed using the Bradford assay (Bio-Rad). 8 Microscopic examination of urine, on a phase contrast microscope, was performed on urine samples that had been centrifuged for 5 minutes at 2000 rpm. The sediment was examined under high power magnification (40×).…”

Section: Renal Functionmentioning

confidence: 99%

Alport syndrome: Proteomic analysis identifies early molecular pathway alterations in Col4a3 knock out mice

et al. 2020

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Aim: Alport syndrome (AS) is the second most common hereditary kidney disease caused by mutations in collagen IV genes. Patients present with microhaematuria that progressively leads to proteinuria and end stage renal disease. Currently, no specific treatment exists for AS. Using mass spectrometry based proteomics, we aimed to detect early alterations in molecular pathways implicated in AS before the stage of overt proteinuria, which could be amenable to therapeutic intervention. Methods: Kidneys were harvested from male Col4a3 −/− knock out and sex and agematched Col4a3 +/+ wild-type mice at 4 weeks of age. Purified peptides were separated by liquid chromatography and analysed by high resolution mass spectrometry. The Cytoscape bioinformatics tool was used for function enrichment and pathway analysis. PPARα expression levels were evaluated by immunofluorescence and immunoblotting. Results: Proteomic analysis identified 415 significantly differentially expressed proteins, which were mainly involved in metabolic and cellular processes, the extracellular matrix, binding and catalytic activity. Pathway enrichment analysis revealed among others, downregulation of the proteasome and PPAR pathways. PPARα protein expression levels were observed to be downregulated in Alport mice, supporting further the results of the discovery proteomics. Conclusion: This study provides additional evidence that alterations in proteins which participate in cellular metabolism and mitochondrial homeostasis in kidney cells are early events in the development of chronic kidney disease in AS. Of note is the dysregulation of the PPAR pathway, which is amenable to therapeutic intervention and provides a new potential target for therapy in AS.

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“…Urine samples were collected every week and stored at -80ºC. Urine albumin was measured using the Bradford assay [24]. Brie y, 4 µL of thawed and diluted urine sample was added to 196 µL of Bradford reagent (Bio-Rad Laboratories, Hercules, CA).…”

Section: Assessment Of Proteinuriamentioning

confidence: 99%

Lupus pathogenesis and autoimmunity are exacerbated by high fat diet–induced obesity in MRL/lpr mice

Zhang

Meng

Shi

et al. 2022

Preprint

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Background: Systemic lupus erythematosus (SLE) is an autoimmune disease characterized by persistent inflammation and autoantibody production, leading to systemic damage. Genetic predisposition and environmental factors such as high fat diet (HFD)/obesity may contribute to lupus development. In this study, we investigated the impact of HFD in lupus pathogenesis and autoimmunity to reveal the pathological link of HFD and SLE using-lupus prone mice.Methods: Sixty MRL/lpr mice were grouped and fed with a regular diet (RD) or HFD. Their body weights were recorded weekly. SLE progression was monitored by skin lesion, urine protein, and titers of anti-dsDNA and antinuclear antibodies (ANA). Mice were euthanized at week 14. Kidney and skin tissue sections were stained with hematoxylin and eosin (H&E), periodic acid–Schiff (PAS), and Masson's trichrome to detect lupus histopathological lesions and were quantified as kidney index and histological skin score. Immune cells in the spleen were identified by immunofluorescence staining and flow cytometry. Student t test or two-way ANOVA was used for statistical analysis.Results: HFD induced a significant increase in mouse body weight by week 5 and continued until week 14 compared to RD (p<0.05 to p<0.01). Skin lesions resembling human discoid lupus were observed as early as week 6 and became severe at week 14 in 55.6% of the HFD group compared to 11.1% of the RD group (p<0.05). Proteinuria occurred from week 11 to week 14 in the HFD group. Splenomegaly was observed in the HFD mice (p<0.05). The HFD group had a higher histological skin score and kidney index than the RD group. An increased trend of anti-dsDNA antibody was detected in the HFD group but no difference in ANA was seen between HFD and RD groups. A significant increase of germinal center B cells and plasma cells was observed in the spleens of HFD mice. The percentage of Tfh cells and the ratio of Tfh/Treg were significantly increased in the HFD group (p<0.05). Conclusion: Our results showed that HFD induced an accelerated and exacerbated lupus development and autoimmunity in MRL/lpr mice. Tfh/Treg cells may be the interconnection of HFD and SLE. Interventions of healthy diet or restoring balance of Tfh/Treg cells may improve both lupus symptoms and outcomes in genetically predisposed SLE patients.

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Determination of urinary peptides in patients with proteinuria

Cited by 8 publications

References 18 publications

Lupus pathogenesis and autoimmunity are exacerbated by high fat diet-induced obesity in MRL/lpr mice

Lupus pathogenesis and autoimmunity are exacerbated by high fat diet-induced obesity in MRL/lpr mice

Alport syndrome: Proteomic analysis identifies early molecular pathway alterations in Col4a3 knock out mice

Lupus pathogenesis and autoimmunity are exacerbated by high fat diet–induced obesity in MRL/lpr mice

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