scholarly journals Upregulation ofα3β1-Integrin in Podocytes in Early-Stage Diabetic Nephropathy

2016 ◽  
Vol 2016 ◽  
pp. 1-7 ◽  
Author(s):  
Kaichiro Sawada ◽  
Masao Toyoda ◽  
Noriko Kaneyama ◽  
Sawako Shiraiwa ◽  
Hitomi Moriya ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Early Stage ◽  
Integrin Expression ◽  
Advanced Stage ◽  
Podocyte Injury ◽  
Expression Levels ◽  
Podocyte Detachment ◽  
Advanced Stages

Background. Podocyte injury plays an important role in the onset and progression of diabetic nephropathy (DN). Downregulation ofα3β1-integrin expression in podocytes is thought to be associated with podocyte detachment from the glomerular basement membrane, although the mechanisms remain obscure. To determine the mechanism of podocyte detachment, we analyzed the expression levels ofα3β1-integrin in podocytes in early and advanced stages of DN.Methods. Surgical specimens from DN patients were examined byin situhybridization, and the expression levels ofα3- andβ1-integrin subunits in glomeruli of early (n=6) and advanced (n=8) stages were compared with those of normal glomeruli (n=5). Heat-sensitive mouse podocytes (HSMP) were cultured with TGF-β1 to reproduce the microenvironment of glomeruli of DN, and the expression levels of integrin subunits and the properties of migration and attachment were examined.Results. Podocytes of early-stage DN showed upregulation ofα3- andβ1-integrin expression while those of advanced stage showed downregulation. Real-time PCR indicated a tendency for upregulation ofα3- andβ1-integrin in HSMP cultured with TGF-β1. TGF-β1-stimulated HSMP also showed enhancedin vitromigration and attachment on collagen substrate.Conclusions. The results suggested that podocyte detachment during early stage of DN is mediated through upregulation ofα3β1-integrin.

Changes in the incidence of cervical tumours by disease stage in a cytology-based screening programme

2019 ◽  
Vol 27 (2) ◽  
pp. 96-104
Author(s):  
Lauro Bucchi ◽  
Silvia Mancini ◽  
Flavia Baldacchini ◽  
Orietta Giuliani ◽  
Alessandra Ravaioli ◽  
...  
Keyword(s):  
Steady State ◽  
Incidence Rate ◽  
Screening Programme ◽  
Early Stage ◽  
Squamous Carcinoma ◽  
Disease Stage ◽  
Adenocarcinoma In Situ ◽  
Advanced Stage ◽  
Tumour Stage

Objectives To report changes in incidence of cervical tumours by disease stage, following the introduction of an organized cytology-based screening programme. Methods An intention-to-screen study of a cytology-based screening programme targeting 1,219,000 women aged 25–64 in northern Italy was carried out. Based on the previously reported trend in total incidence of cervical cancer, the study period 1995–2014 was divided into 1995–1996 (pre-screening, or reference, years), 1997–1998 (screening implementation phase), 1999–2006 (transition phase, when incidence decreased), and 2007–2014 (steady-state phase, when incidence stabilized again). Tumour stage was categorized as preinvasive (cervical intraepithelial neoplasia grade 3 (CIN3) and adenocarcinoma in situ), early (pT1a), advanced (pT1b or greater, ypT), and unknown (pT1 not otherwise specified, pTx, missing information). Average annual incidence rates observed in each phase were compared with the expected (reference) rates, using the incidence rate ratio, calculated with a Poisson regression model. Results In the steady-state phase, incidence rate ratios were: CIN3, 1.55 (95% confidence interval, 1.41–1.70); early-stage squamous carcinoma, 0.49 (0.36–0.67); advanced-stage squamous carcinoma, 0.44 (0.33–0.57); unknown-stage squamous carcinoma, 0.69 (0.48–0.99); adenocarcinoma in situ, 1.44 (0.72–2.88); early-stage adenocarcinoma, 2.65 (0.82–8.53); advanced-stage adenocarcinoma, 1.03 (0.56–1.91); and unknown-stage adenocarcinoma, 0.46 (0.23–0.92). Conclusions After stabilization, changes in incidence by tumour stage included a 55% increase for CIN3 and a 50–55% decrease both for early- and advanced-stage squamous carcinoma, but no significant changes for glandular tumours. These data will serve to quantify the incremental impact of the implementation of human papillomavirus-based screening, introduced in 2015.

An Investigation of Radial Reflux in an Isolated Peripheral Canine Vein Segment

1995 ◽  
Vol 10 (3) ◽  
pp. 115-121 ◽  
Author(s):  
T. P. Crotty
Keyword(s):  
Turbulent Flow ◽  
Varicose Veins ◽  
Early Stage ◽  
Vein Segment ◽  
Adrenergic Activity ◽  
Conventional Histology ◽  
University Department

Objective: To investigate the effects of flow from the vein lumen into the vasa venarum (radial reflux) and the factors regulating it. Design: In vivo and in vitro animal study. Setting: University Department of Physiology. Materials: Segments of canine vein were perfused in vitro ( n = 84) and in situ ( n = 60). Main outcome measures: Specimens were studied by conventional histology using light microscopy. Results: Radial reflux caused dilatation of constricted segments and when associated with turbulent flow resulted in the formation of acute short-lived varices, marked dilatation of sinusoidal venules at the medioadventitial junction of the segments and of sinusoidal venules in the paradventitial tissue. Conclusion: During laminar flow radial reflux operated as a feedback to regulate venoconstrictor tone. During turbulent flow it caused more widespread vascular changes and inhibited adrenergic activity. The changes are of particular interest because they mimicked those that characterize the early stage of varicose veins.

scholarly journals Automated image analysis of a glomerular injury marker desmin in spontaneously diabetic Torii rats treated with losartan

2014 ◽  
Vol 222 (1) ◽  
pp. 43-51 ◽  
Author(s):  
Tetsuhiro Kakimoto ◽  
Kinya Okada ◽  
Yoshihiro Hirohashi ◽  
Raissa Relator ◽  
Mizue Kawai ◽  
...  
Keyword(s):  
Image Analysis ◽  
Diabetic Nephropathy ◽  
Early Stage ◽  
Automated Image Analysis ◽  
Angiotensin Ii Receptor Blocker ◽  
Analysis Method ◽  
Glomerular Injury ◽  
Image Analysis Method ◽  
Podocyte Injury

Diabetic nephropathy is a major complication in diabetes and a leading cause of end-stage renal failure. Glomerular podocytes are functionally and structurally injured early in diabetic nephropathy. A non-obese type 2 diabetes model, the spontaneously diabetic Torii (SDT) rat, is of increasing preclinical interest because of its pathophysiological similarities to human type 2 diabetic complications including diabetic nephropathy. However, podocyte injury in SDT rat glomeruli and the effect of angiotensin II receptor blocker treatment in the early stage have not been reported in detail. Therefore, we have evaluated early stages of glomerular podocyte damage and the beneficial effect of early treatment with losartan in SDT rats using desmin as a sensitive podocyte injury marker. Moreover, we have developed an automated, computational glomerulus recognition method and illustrated its specific application for quantitatively studying glomerular desmin immunoreactivity. This state-of-the-art method enabled automatic recognition and quantification of glomerular desmin-positive areas, eliminating the need to laboriously trace glomerulus borders by hand. The image analysis method not only enabled assessment of a large number of glomeruli, but also clearly demonstrated that glomerular injury was more severe in the juxtamedullary region than in the superficial cortex region. This applied not only in SDT rat diabetic nephropathy but also in puromycin aminonucleoside-induced nephropathy, which was also studied. The proposed glomerulus image analysis method combined with desmin immunohistochemistry should facilitate evaluations in preclinical drug efficacy studies as well as elucidation of the pathophysiology of diabetic nephropathy.

scholarly journals METTL14 aggravates podocyte injury and glomerulopathy progression through N6-methyladenosine-dependent downregulating of Sirt1

2021 ◽  
Vol 12 (10) ◽  
Author(s):  
Zhihui Lu ◽  
Hong Liu ◽  
Nana Song ◽  
Yiran Liang ◽  
Jiaming Zhu ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Posttranscriptional Regulation ◽  
Kidney Diseases ◽  
Luciferase Reporter ◽  
Podocyte Injury ◽  
Glomerular Function ◽  
Apoptosis And Inflammation ◽  
Dual Luciferase Reporter Assay

AbstractPodocytes are known to play a determining role in the progression of proteinuric kidney disease. N6-methyladenosine (m6A), as the most abundant chemical modification in eukaryotic mRNA, has been reported to participate in various pathological processes. However, its role in podocyte injury remains unclear. In this study, we observed the elevated m6A RNA levels and the most upregulated METTL14 expression in kidneys of mice with adriamycin (ADR) and diabetic nephropathy. METTL14 was also evidently increased in renal biopsy samples from patients with focal segmental glomerulosclerosis (FSGS) and diabetic nephropathy and in cultured human podocytes with ADR or advanced glycation end product (AGE) treatment in vitro. Functionally, we generated mice with podocyte-specific METTL14 deletion, and identified METTL14 knockout in podocytes improved glomerular function and alleviated podocyte injury, characterized by activation of autophagy and inhibition of apoptosis and inflammation, in mice with ADR nephropathy. Similar to the results in vivo, knockdown of METTL14 facilitated autophagy and alleviated apoptosis and inflammation in podocytes under ADR or AGE condition in vitro. Mechanically, we identified METTL14 knockdown upregulated the level of Sirt1, a well-known protective deacetylase in proteinuric kidney diseases, in podocytes with ADR or AGE treatment. The results of MeRIP-qPCR and dual-luciferase reporter assay indicated METTL14 promoted Sirt1 mRNA m6A modification and degradation in injured podocytes. Our findings suggest METTL14-dependent RNA m6A modification contributes to podocyte injury through posttranscriptional regulation of Sirt1 mRNA, which provide a potential approach for the diagnosis and treatment of podocytopathies.

scholarly journals Islet Transplantation Reverses Podocyte Injury in Diabetic Nephropathy or Induced by High Glucose via Inhibiting RhoA/ROCK/NF-κB Signaling Pathway

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Chongchu Huang ◽  
Yi Zhou ◽  
Hongjian Huang ◽  
Yushu Zheng ◽  
Lijun Kong ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Signaling Pathways ◽  
Signaling Pathway ◽  
Islet Transplantation ◽  
High Glucose ◽  
Islet Cells ◽  
Inflammatory Factors ◽  
Efficacy Evaluation ◽  
Podocyte Injury ◽  
Expression Levels

Objective. Abnormal signaling pathways play a crucial role in the mechanisms of podocyte injury in diabetic nephropathy. They also affect the recovery of podocytes after islet transplantation (IT). However, the specific signaling abnormalities that affect the therapeutic effect of IT on podocytes remains unclear. The purpose of this study was to assess whether the RhoA/ROCK/NF-κB signaling pathway is related to podocyte restoration after IT. Methods. A mouse model of diabetic nephropathy was established in vivo using streptozotocin. The mice were then subsequently reared for 4 weeks after islet transplantation to determine the effect of IT. Islet cells, CCG-1423 (RhoA Inhibitor), and fasudil (ROCK inhibitor) were then cocultured with podocytes in vitro to assess their protective effects on podocyte injury induced by high glucose (HG). Protein expression levels of RhoA, ROCK1, synaptopodin, IL-6, and MCP-1 in kidney tissues were then measured using immunohistochemistry and Western blotting techniques. Results. Islet transplantation reduced the expression levels of RhoA/ROCK1 and that of related inflammatory factors such as IL-6 and MCP-1 in the kidney podocytes of diabetic nephropathy. In the same line, islet cells reduced the expression of RhoA, ROCK1, and pp65 in immortalized podocytes under high glucose (35.0 mmol/L glucose) conditions. Conclusions. Islet transplantation can reverse podocyte injury in diabetes nephropathy by inhibiting the RhoA/ROCK1 signaling pathway. Islet cells have a strong protective effect on podocytes treated with high glucose (35.0 mmol/L glucose). Discovery of signaling pathways affecting podocyte recovery is helpful for individualized efficacy evaluation and targeted therapy of islet transplantation patients.

scholarly journals MicroRNA-27a targets Sfrp1 to induce renal fibrosis in diabetic nephropathy by activating Wnt/β-Catenin signalling

2020 ◽  
Vol 40 (6) ◽  
Author(s):  
MingJun Shi ◽  
PingPing Tian ◽  
ZhongQiang Liu ◽  
Fan Zhang ◽  
YingYing Zhang ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
High Glucose ◽  
Renal Fibrosis ◽  
Negative Control ◽  
Luciferase Reporter ◽  
Mrna Levels ◽  
Expression Levels ◽  
Stage Renal Disease

Abstract Diabetic nephropathy (DN) commonly causes end-stage renal disease (ESRD). Increasing evidence indicates that abnormal miRNA expression is tightly associated with chronic kidney disease (CKD). This work aimed to investigate whether miR-27a can promote the occurrence of renal fibrosis in DN by suppressing the expression of secreted frizzled-related protein 1 (Sfrp1) to activate Wnt/β-catenin signalling. Therefore, we assessed the expression levels of miR-27a, Sfrp1, Wnt signalling components, and extracellular matrix (ECM)-related molecules in vitro and in vivo. Sfrp1 was significantly down-regulated in a high-glucose environment, while miR-27a levels were markedly increased. A luciferase reporter assay confirmed that miR-27a down-regulated Sfrp1 by binding to the 3′ untranslated region directly. Further, NRK-52E cells under high-glucose conditions underwent transfection with miR-27a mimic or the corresponding negative control, miR-27a inhibitor or the corresponding negative control, si-Sfrp1, or combined miR-27a inhibitor and si-Sfrp1. Immunoblotting and immunofluorescence were performed to assess the relative expression levels of Wnt/β-catenin signalling and ECM components. The mRNA levels of Sfrp1, miR-27a, and ECM-related molecules were also detected by quantitative real-time PCR (qPCR). We found that miR-27a inhibitor inactivated Wnt/β-catenin signalling and reduced ECM deposition. Conversely, Wnt/β-catenin signalling was activated, while ECM deposition was increased after transfection with si-Sfrp1. Interestingly, miR-27a inhibitor attenuated the effects of si-Sfrp1. We concluded that miR-27a down-regulated Sfrp1 and activated Wnt/β-catenin signalling to promote renal fibrosis.

scholarly journals New insights on microRNAs in diabetic nephropathy: potential biomarkers for diagnosis and therapeutic targets

2017 ◽  
Vol 20 (1) ◽  
pp. 42-50 ◽  
Author(s):  
Elena Sergeevna Kamyshova ◽  
Irina Nikolaevna Bobkova ◽  
Irina Mikhailovna Kutyrina
Keyword(s):  
Diabetic Nephropathy ◽  
Early Detection ◽  
Early Stage ◽  
Biological Fluids ◽  
Renal Tissue ◽  
In Vivo Models ◽  
New Biomarkers

Diabetic nephropathy (DN) is a severe complication of diabetes mellitus associated with the progressive deterioration of renal function. Although microalbuminuria is considered as a gold standard for DN diagnosis, it has limited predictive powers and specificity as a diagnostic tool for the early stage of DN. Therefore, new biomarkers are required for the early detection of DN. Studies using in vitro and in vivo models of DN have revealed an important role of microRNAs (miRNAs), short non-coding RNAs that modulate physiological and pathological processes by inhibiting target gene expression, in DN development. Recent studies have shown that the dysregulation of miRNAs, which is associated with the key features of DN, such as the mesangial expansion and accumulation of extracellular matrix proteins, is related to fibrosis and glomerular dysfunction. Thus, the up- and downregulation of miRNA expression in the renal tissue or biological fluids, including urine, may represent new biomarkers for the diagnosis and monitoring of DN progression. In this review, we highlight the significance of miRNAs as biomarkers for the early detection of DN and emphasise their potential role as a therapeutic target.

scholarly journals In-Vivo Toxicity Studies and In-Vitro Inactivation of SARS-CoV-2 by Povidone-iodine In-situ Gel Forming Formulations

2020 ◽  
Author(s):  
Bo Liang ◽  
Xudong Yuan ◽  
Gang Wei ◽  
Wei Wang ◽  
Ming Zhang ◽  
...  
Keyword(s):  
Povidone Iodine ◽  
Early Stage ◽  
Etiologic Agent ◽  
Dependent Manner ◽  
Forming Technology ◽  
Long Acting ◽  
Dose Dependent

AbstractTo curb the spread of SARS-CoV-2, the etiologic agent of the COVID-19 pandemic, we characterize the virucidal activity of long-acting Povidone Iodine (PVP-I) compositions developed using an in-situ gel forming technology. The PVP-I gel forming nasal spray (IVIEW-1503) and PVP-I gel forming ophthalmic eye drop (IVIEW-1201) rapidly inactivated SARS-CoV-2, inhibiting the viral infection of VERO76 cells. No toxicity was observed for the PVP-I formulations. Significant inactivation was noted with preincubation of the virus with these PVP-I formulations at the lowest concentrations tested. It has been demonstrated that both PVP-I formulations can inactivate SARS-CoV-2 virus efficiently in both a dose-dependent and a time-dependent manner. These results suggest IVIEW-1503 and IVIEW-1201 could be potential agents to reduce or prevent the transmission of the virus through the nasal cavity and the eye, respectively. Further studies are needed to clinically evaluate these formulations in early-stage COVID-19 patients.

scholarly journals Study on the Role of Cytc in Response to BmNPV Infection in Silkworm, Bombyx mori (Lepidoptera)

2019 ◽  
Vol 20 (18) ◽  
pp. 4325 ◽  
Author(s):  
Xue-Yang Wang ◽  
Kang-Hui Wu ◽  
Hui-Lin Pang ◽  
Ping-Zhen Xu ◽  
Mu-Wang Li ◽  
...  
Keyword(s):  
Bombyx Mori ◽  
Early Stage ◽  
Malpighian Tubule ◽  
Infection Process ◽  
Insect Vector ◽  
Apoptosis Pathway ◽  
Expression Levels ◽  
Mitochondrial Apoptosis Pathway

Bombyx mori nucleopolyhedrovirus (BmNPV) is one of the primary pathogens of the silkworm. Cytochrome c (cytc) showed a significant response to BmNPV infection in our previous transcriptome study. However, little is known about the role of Bombyx mori cytc (Bmcytc) in resistance to BmNPV infection. In this study, the expression levels analysis of Bmcytc showed stable expression levels in selected tissues of the resistant strain AN following BmNPV infection, while there was downregulation in the susceptible strain p50, except in the malpighian tubule. To further study the role of Bmcytc in viral infection, Bmcytc was knocked down with siRNA in vitro, resulting in significant downregulation of selected downstream genes of the mitochondrial pathway, including Bmapaf, Bmcaspase-Nc, and Bmcaspase-1; this was also confirmed by overexpression of Bmcytc using the pIZT/V5-His-mCherry insect vector, except Bmcaspase-1. Moreover, knockdown of Bmcytc significantly promoted the infection process of BmNPV in vitro, while the infection was inhibited by overexpression of Bmcytc at the early stage and subsequently increased rapidly. Based on these results, we concluded that Bmcytc plays a vital role in BmNPV infection by regulating the mitochondrial apoptosis pathway. Our work provides valuable data for the clarification of the mechanism of silkworm resistance to BmNPV infection.

P0989VDR/ATG3 AXIS REGULATES SLIT DIAGRAM TO TIGHT JUNCTION TRANSITION VIA P62-MEDIATED AUTOPHAGY PATHWAY IN DIABETIC NEPHROPATHY

2020 ◽  
Vol 35 (Supplement_3) ◽  
Author(s):  
Jingyi Qian ◽  
Bin Wang ◽  
Bicheng Liu
Keyword(s):  
Diabetic Nephropathy ◽  
Tight Junction ◽  
Degradation Pathway ◽  
Serum Starvation ◽  
Autophagic Flux ◽  
Vdr Gene ◽  
Podocyte Injury ◽  
Autophagy Pathway

Abstract Background and Aims Vitamin D receptor (VDR) loss, slit diagram (SD) to tight junction (TJ) transition and impaired autophagic flux contribute to podocyte injury in diabetic nephrology. This study aims to examine the effect and mechanism of VDR on autophagic flux and SD-TJ transition in diabetic nephropathy. Method Renal biopsy tissues from DN patients at stage IIa, IIb, III, IV and patients with minimal lesions were used to evaluate the expression of VDR, autophagic flux and SD-TJ transition glomeruli. In vitro, cultured podocytes were treated with serum starvation (SS), autophagic inhibitors (3-methyladenine 3-MA or chloroquine CQ) to determine the degradation pathway of TJ marker ZO-1. Meanwhile, db/db mice and STZ-induced rats were used to explore the therapeutic effect and mechanism of VDR agonist in diabetic nephropathy. Results SD-TJ transition between foot processes could be observed under electron microscopy in DN patients at all stages, whereas foot processes were separated by the filtration slit and appeared to be single cross-strands in the normal glomeruli. There was a trend of increasing expression of autophagic marker p62 and ZO-1 and the expression of p62 is positively correlated with the changes of ZO-1 in the glomeruli of DN patients. In vitro, inhibiting autophagy with 3-MA and CQ resulted in the accumulation of ZO-1 in cultured podocytes. In addition, Co-IP experiments further convinced the interaction between p62 and ZO-1, which was enhanced by the activation of autophagy. Podocytes apoptosis and the activity of caspase 3 and caspase 8 were significantly increased in the presence of 3-MA or CQ, while these effects were rescued by silencing p62. According to VDR gene expression data in GEO database, VDR expression was decreased in diabetic nephropathy patients compared with normal people. Knocking down VDR lowered the expression of atg3 and leaded to the blockage of autophagy, which could be reversed by over-expressing Atg3. Podocytes treated with high glucose resulted in the decrease of VDR and Atg3, impaired autophagic flux and aggravated podocytes injury. However, VDR agonist treatment partially reversed all the changes. In vivo, db/db mice and STZ-induced rats (DN animal models) exhibited SD-TJ transition, massive proteinuria, decreased expression of VDR and podocin and the increased accumulation of p62 and ZO-1, all of which could be partially reversed by VDR agonist. Conclusion VDR loss contributed to the impairment of autophagic flux and SD-TJ transition via down-regulation Atg3 in diabetic nephropathy. Here, we identified a new mechanism and evidence for VDR agonist to treat diabetic nephropathy.

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