Beta-2-microglobulin (B2M) expression in the urinary sediment correlates with clinical markers of kidney disease in patients with type 1 diabetes

Monteiro, Maria Beatriz; Thieme, Karina; Santos‐Bezerra, Daniele Pereira; Queiroz, Márcia Silva; Woronik, Viktoria; Passarelli, Marisa; Machado, Ubiratan Fabres; Giannella-Neto, Daniel; Oliveira‐Souza, Maria; Corrêa-Giannella, Maria Lúcia

doi:10.1016/j.metabol.2016.02.012

Cited by 26 publications

(18 citation statements)

References 27 publications

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“…For example, in diabetic kidney, B2m contributes to diabetic tubulopathy and might not be a good RG for the kidney [51]. GAPDH has been reported to contribute to neurological disorder such as Huntington and Alzheimer diseases [52] which indicates GAPDH might not be an optimal candidate gene in hypothalamus.…”

Section: Discussionmentioning

confidence: 99%

Selection of suitable endogenous reference genes for qPCR in kidney and hypothalamus of rats under testosterone influence

et al. 2017

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Real-time quantitative PCR (qPCR) is the most reliable and accurate technique for analyses of gene expression. Endogenous reference genes are being used to normalize qPCR data even though their expression may vary under different conditions and in different tissues. Nonetheless, verification of expression of reference genes in selected studied tissue is essential in order to accurately assess the level of expression of target genes of interest. Therefore, in this study, we attempted to examine six commonly used reference genes in order to identify the gene being expressed most constantly under the influence of testosterone in the kidneys and hypothalamus. The reference genes include glyceraldehyde-3-phosphate dehydrogenase (GAPDH), actin beta (ACTB), beta-2 microglobulin (B2m), hypoxanthine phosphoribosyltransferase 1 (HPRT), peptidylprolylisomerase A (Ppia) and hydroxymethylbilane synthase (Hmbs). The cycle threshold (Ct) value for each gene was determined and data obtained were analyzed using the software programs NormFinder, geNorm, BestKeeper, and rank aggregation. Results showed that Hmbs and Ppia genes were the most stably expressed in the hypothalamus. Meanwhile, in kidneys, Hmbs and GAPDH appeared to be the most constant genes. In conclusion, variations in expression levels of reference genes occur in kidneys and hypothalamus under similar conditions; thus, it is important to verify reference gene levels in these tissues prior to commencing any studies.

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Section: Discussionmentioning

confidence: 99%

Selection of suitable endogenous reference genes for qPCR in kidney and hypothalamus of rats under testosterone influence

et al. 2017

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“…Several studies have shown that the phenotype of cultured GMCs caused by HG environment mimics the pathophysiological changes in diabetic nephropathy [24]. Gene variations also critically contribute to the renal dysfunction in diabetes [25, 26]. We found that high concentration DBD extract significantly inhibited GMCs proliferation at all three time-points (24, 48 and 72 h), while medium concentration DBD extract only inhibited GMCs proliferation at 72 h. Low concentration DBD extract did not seem to inhibit proliferation of GMCs.…”

Section: Discussionmentioning

confidence: 99%

Effects of Dangguibuxue decoction on rat glomerular mesangial cells cultured under high glucose conditions

Ren

Zhang

Wang

et al. 2017

BMC Complement Altern Med

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BackgroundDysfunction of glomerular mesangial cells (GMCs) plays an important role in pathogenesis of diabetic nephropathy. Here, we investigated the effects of Dangguibuxue decoction (DBD), an herbal traditional Chinese medicinal (TCM) formula composed of Astragali Radix and Angelicae Sinensis Radix, on GMC proliferation and fibrogenesis under high-glucose (HG) conditions.MethodsSixty male Sprague Dawley rats were divided into 5 groups and administered intragastric 0.9% saline, low concentration DBD (DBD-L, 1.75 g/kg/d), middle concentration DBD (DBD-M, 3.5 g/kg/d), high concentration DBD (DBD-H, 7.0 g/kg/d) and gliclazide (GL, 2 mg/kg/d), respectively, for 1 week, and then their sera were obtained. Rat mesangial cells (HBZY-1 cells) were treated with these sera under HG condition (30 mmol/L).ResultsThe proliferation of GMCs under HG conditions was significantly greater than that under normal glucose condition. Low concentration DBD (DBD-L) inhibited proliferation of GMCs after 72-h incubation (P < 0.01), while high concentration DBD (DBD-H) inhibited GMCs proliferation at 24, 48 and 72 time points (P < 0.01). There was no significant difference between the inhibitory effect of DBD-H and GL sera on GMC proliferation (P > 0.05). Furthermore, all concentrations of DBD (DBD-L, DBD-M and DBD-H) significantly decreased the protein expression of α-SMA(α-smooth muscle actin) (P < 0.01), an indicator of interstitial fibrosis of GMCs. Finally, DBD-L, DBD-M, DBD-H sera obviously inhibited the increase of HYP (hydroxyproline)secretion under HG condition (P < 0.01).ConclusionOur results demonstrate an inhibitory effect of DBD extract on proliferation and fibrogenesis of GMCs under HG conditions. The potential role of DBD in the treatment of diabetic neuropathy merits further investigation.

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“…All patients were receiving intensive insulin therapy. At the time of recruitment, urine samples were collected as previously described (5,6) and later used for gene expression analyses. Briefly, urine specimens were collected in sterile RNAse-free flasks.…”

Section: Participantsmentioning

confidence: 99%

“…Gene expression analyses were performed with 10 ng of cDNA/sample, in duplicates, with the use of Taqman assays in a StepOne plus Real-Time PCR System (ThermoFisher Scientific, Carlsbad, CA). The relative mRNA abundance was calculated using the 2-Ct method (13) and E74-Like Factor 1 (ELF1) was used as reference gene (5). Taqman assays are listed in Supplemental Table 1.…”

Section: Qrt-pcr Validationmentioning

confidence: 99%

“…Besides being an organ-specific sample, urine is an easily obtainable material, without the need of invasive procedures. We previously described the expression of markers of proximal tubule epithelial cells and podocytes in the human urine sediment while attempting to correlate clinical markers of kidney disease with expression of target genes in this biological material (5). The same approach was taken by other research groups (3) showing the use of urine as a translatable strategy to study markers of kidney injury without the need of invasive interventions.…”

Section: Introductionmentioning

confidence: 99%

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Urinary Sediment Transcriptomic and Longitudinal Data to Investigate Renal Function Decline in Type 1 Diabetes

et al. 2020

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Beta-2-microglobulin (B2M) expression in the urinary sediment correlates with clinical markers of kidney disease in patients with type 1 diabetes

Cited by 26 publications

References 27 publications

Selection of suitable endogenous reference genes for qPCR in kidney and hypothalamus of rats under testosterone influence

Selection of suitable endogenous reference genes for qPCR in kidney and hypothalamus of rats under testosterone influence

Effects of Dangguibuxue decoction on rat glomerular mesangial cells cultured under high glucose conditions

Urinary Sediment Transcriptomic and Longitudinal Data to Investigate Renal Function Decline in Type 1 Diabetes

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