scholarly journals Down-regulation of Risa Improves Podocyte Injury by Enhancing Autophagy in Diabetic Nephropathy

2021 ◽  
Author(s):  
Peipei Su ◽  
Dongwei Liu ◽  
Sijie Zhou ◽  
Hang Chen ◽  
Xianming Wu ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Poor Prognosis ◽  
Kidney Tissue ◽  
Podocyte Injury ◽  
Tissue Samples ◽  
Adeno Associated Virus ◽  
Qrt Pcr ◽  
Tail Vein Injection ◽  
Transmission Electron ◽  
Pas Staining

Abstract BackgroundLncRNA AK044604 (Risa), Sirt1 and GSK3β are autophagy related genes that can play important roles in diabetic nephropathy (DN). In this study, we sought to explore the effect of Risa on Sirt1/GSK3β-induced podocyte injury.MethodsTransgenic db/db mice were fed and injected with Risa inhibition of adeno-associated virus by tail vein injection, as well as intraperitoneally injected with LiCl. Blood, urine, kidney tissue samples, and clinical data were collected at different time points. Immortalized mouse podocyte cells (MPCs) were cultured and treated with Risa inhibition of lentivirus, EX-527, and LiCl. MPCs were collected under different stimulations. The effects of Risa on podocyte autophagy were examined by qRT-PCR, Western blot analysis, transmission electron microscope, PAS staining, and immunofluorescence staining. ResultsRisa and activated GSK3β were overexpressed, but Sirt1 decreased in Renal tissues of DN mice and high-glucose-treated MPCs and correlated with poor prognosis. Risa overexpression attenuated Sirt1-mediated downstream autophagy levels and aggravated the injury of podocytes by inhibiting the expression of Sirt1. In contrast, Risa inhibition enhanced Sirt1-induced autophagy and attenuated podocyte injury, but this effect could be abrogated by EX-527, suggesting that Risa overexpression aggravated podocyte injury by decreasing autophagy. ConclusionsRisa inhibits autophagy by regulating the Sirt1/GSK3β axis and thereby aggravates podocyte injury in DN. Risa may serve as a therapeutic target for the treatment of DN.

scholarly journals Artificially Cultivated Ophiocordyceps sinensis Alleviates Diabetic Nephropathy and Its Podocyte Injury via Inhibiting P2X7R Expression and NLRP3 Inflammasome Activation

2018 ◽  
Vol 2018 ◽  
pp. 1-16 ◽  
Author(s):  
Chao Wang ◽  
Xiao-xia Hou ◽  
Hong-liang Rui ◽  
Li-jing Li ◽  
Jing Zhao ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Protein Expression ◽  
Nlrp3 Inflammasome ◽  
Kidney Tissue ◽  
Therapeutic Effects ◽  
Low Grade ◽  
Podocyte Injury ◽  
Ophiocordyceps Sinensis ◽  
Caspase 1 ◽  
Mrna And Protein Expression

Background/Aims. It is known that chronic low-grade inflammation contributes to the initiation and development of both diabetes and diabetic nephropathy (DN), so we designed this study to investigate the role of P2X7R and NLRP3 inflammasome in DN pathogenesis and the antagonistic effects of artificially cultivated Ophiocordyceps sinensis (ACOS). Methods. A rat model of DN caused by high-fat-diet feeding and low-dose streptozotocin injection and a mouse podocyte injury model induced by high-glucose (HG) stimulation were established, and the intervention effects of ACOS on them were observed. The biological parameters of serum and urine and the pathological manifestations of kidney tissue were examined. The expression of mRNA and protein of P2X7R and NLRP3 inflammasome (NLRP3, ASC, and caspase-1) and downstream effectors (IL-1β and IL-18), as well as podocyte-associated molecules, was determined by real-time quantitative PCR and Western blot assay, respectively. Results. The DN rats showed to have developed insulin resistance, elevated fasting blood glucose, increased urinary protein excretion, and serum creatinine level as well as corresponding glomerular pathological alterations including podocyte damages. ACOS significantly antagonized the above changes. The experiments in vivo and in vitro both displayed that the mRNA and protein expression of P2X7R, NLRP3, ASC, caspase1 (procaspase-1 mRNA in the gene level and active caspase-1 subunit P10 in the protein level), IL-1β, and IL-18 was significantly upregulated and the mRNA and protein expression of podocyte-associated molecules was significantly changed (downregulation of nephrin, podocin, and WT-1 expression and upregulation of desmin expression) indicating podocyte injury in the kidney tissue of DN rats and in the HG-stressed mouse podocytes, respectively. ACOS also significantly antagonized all the above changes. Conclusion. Our research work suggests that P2X7R and NLRP3 inflammasome are involved in the pathogenesis of DN, and ACOS can effectively inhibit the high expression of P2X7R and the activation of NLRP3 inflammasome, which may contribute to the therapeutic effects of Ophiocordyceps sinensis.

Soluble Flt-1 gene therapy ameliorates albuminuria but accelerates tubulointerstitial injury in diabetic mice

2010 ◽  
Vol 298 (3) ◽  
pp. F609-F616 ◽  
Author(s):  
Tomoki Kosugi ◽  
Takahiro Nakayama ◽  
Qiuhong Li ◽  
Vince A. Chiodo ◽  
Li Zhang ◽  
...  
Keyword(s):  
Gene Therapy ◽  
Diabetic Nephropathy ◽  
Diabetic Mice ◽  
Tubulointerstitial Injury ◽  
Podocyte Injury ◽  
Adeno Associated Virus ◽  
Translational Study ◽  
Renal Histology ◽  
Progression Of Renal Disease ◽  
Different Doses

VEGF is recognized as a major mediator in the development of diabetic nephropathy. Soluble Flt-1 (sFlt-1) is the endogenous inhibitor of VEGF, and recently genetic overexpression of sFlt-1 in the podocyte was shown to be protective in murine diabetic nephropathy. In this study, we performed a translational study to determine whether an intramuscular gene transfer of sFlt-1 can prevent the progression of renal disease in diabetic db/db mice. Adeno-associated virus-1 (AAV1) encoding human sFlt-1 in two different doses was intramuscularly administrated in db/db and wild-type mice. The sFlt-1-AAV1 treatment significantly increased serum sFlt-1 level at 4 and 8 wk. A dose that was developed in this study caused minimal abnormalities in normal mice but reduced albuminuria in diabetic db/db mice. In renal histology, sFlt-1 treatment at this dose had minimal effects on mesangial expansion in diabetic mice, whereas podocyte injury was significantly improved, at 8 wk. Unfortunately, tubulointerstitial injury was markedly exacerbated by sFlt-1 treatment in association with a reduction in endogenous VEGF expression and peritubular capillary loss. In conclusion, gene therapy with sFlt-1-AAV1 protects podocytes but accelerates tubulointerstitial injury in diabetic db/db mice. These data suggest systemic overexpression of sFlt-1 will not likely be useful for treating diabetic nephropathy.

scholarly journals Baicalin improves podocyte injury in rats with diabetic nephropathy by inhibiting PI3K/Akt/mTOR signaling pathway

Open Medicine ◽  
2021 ◽  
Vol 16 (1) ◽  
pp. 1286-1298
Author(s):  
Yi Ou ◽  
Wenjuan Zhang ◽  
Shaopeng Chen ◽  
Haihua Deng
Keyword(s):  
Diabetic Nephropathy ◽  
Signaling Pathway ◽  
Periodic Acid ◽  
Fasting Blood Glucose ◽  
Kidney Tissue ◽  
Mtor Signaling ◽  
Dependent Manner ◽  
Mtor Signaling Pathway ◽  
Podocyte Injury ◽  
Treatment Groups

Abstract Objective To investigate the effect of baicalin on diabetic nephropathy (DN) rats and podocytes and its mechanism. Methods The rat models with DN were established by high-fat and high-sugar diet and intraperitoneal injection of streptozotocin. The fasting blood glucose (FBG) and weight of rats in each group were measured at 0, 1, 2, 3, and 4 weeks. Their biochemical indicators, expression of inflammatory, and antioxidant factors were measured using an automatic biochemical analyzer together with ELISA. Hematoxylin–eosin staining and periodic acid-schiff staining were used to observe the morphological changes in the kidneys of rats in each group. Finally, the expressions of related molecules and PI3K/Akt/mTOR signaling pathway proteins in renal tissues and podocytes were examined by qRT-PCR and Western blot. Results Compared with the DN group, the FBG and weight, serum creatinine, blood urea nitrogen, total cholesterol, triacylglycerol, microalbumin, and albumin/creatinine ratio were all significantly decreased in the Baicalin treatment groups in a concentration-dependent manner. The levels of inflammatory factors in kidney tissue and podocytes were decreased. In addition, the activities of lactate dehydrogenase and malondialdehyde in tissue were decreased, while the superoxide dismutase was increased. The pathological sections showed that glomerular atrophy and glomerular basement membrane thickening caused by hyperglycemia were improved in the Baicalin treatment groups. Meanwhile, baicalin inhibited the downregulation of Nephrin and Podocin expressions and upregulation of Desmin expression caused by DN, and inhibited the expressions of p-PI3K, p-Akt, and p-mTOR proteins. Conclusion Baicalin slows down podocyte injury caused by DN by inhibiting the activity of PI3K/Akt/mTOR signaling pathway.

scholarly journals Silencing of miR-150-5p Ameliorates Diabetic Nephropathy by Targeting SIRT1/p53/AMPK Pathway

2021 ◽  
Vol 12 ◽  
Author(s):  
Wenmin Dong ◽  
Huiqian Zhang ◽  
Cheng Zhao ◽  
Yun Luo ◽  
Ying Chen
Keyword(s):  
Diabetic Nephropathy ◽  
Kidney Tissue ◽  
Protective Role ◽  
Sirtuin 1 ◽  
Podocyte Injury ◽  
Stage Renal Disease ◽  
End Stage ◽  
Amp Activated Protein Kinase

Diabetic nephropathy (DN) is a common complication of diabetes and an important cause of end-stage renal disease. Increasing evidence suggests that microRNAs (miRNAs) regulate the development of DN. In a preliminary study, high levels of miR-150-5p were detected in the serum and urine of patients with DN. Consequently, we investigated the effect and mechanism of action of miR-150-5p in DN in vitro and in vivo. Our results showed that inhibition of miR-150-5p reversed high glucose-induced podocyte injury and Streptozocin (STZ)-induced diabetic nephropathy in mice. Further analysis revealed that miR-150-5p targeted the 3′ untranslated region (UTR) of sirtuin 1 (SIRT1), consequently decreasing SIRT1 levels in podocytes. Importantly, we found that the silencing of miR-150-5p promoted the interaction between SIRT1 and p53, causing the suppression of p53 acetylation in podocytes and kidney tissue. This resulted in the stimulation of AMP-activated protein kinase (AMPK)-dependent autophagy. In conclusion, our study demonstrated that the silencing of miR-150-5p played a reno-protective role in DN mice through targeting SIRT1.

scholarly journals Inhibition of miRNA-155 Alleviates High Glucose-Induced Podocyte Inflammation by Targeting SIRT1 in Diabetic Mice

2021 ◽  
Vol 2021 ◽  
pp. 1-11
Author(s):  
Xiaolei Wang ◽  
Yanbin Gao ◽  
Wenming Yi ◽  
Yu Qiao ◽  
Hao Hu ◽  
...  
Keyword(s):  
Diabetic Nephropathy ◽  
Kidney Tissue ◽  
Luciferase Assay ◽  
Inflammatory Factors ◽  
Luciferase Reporter ◽  
Inflammatory Factor ◽  
Podocyte Injury ◽  
Markers Of Inflammation ◽  
Inflammatory Molecule ◽  
Molecule Production

Objective. Microinflammation plays a crucial role in podocyte dysfunction in diabetic nephropathy, but its regulatory mechanism is still unclear. This study is aimed at discussing the mechanisms underlying the effect of miRNA-155 on podocyte injury to determine its potential as a therapeutic target. Methods. Cultured immortalized mouse podocytes and diabetic KK-Ay mice models were treated with a miR-155 inhibitor. Western blotting, real-time PCR, ELISA, immunofluorescence, and Luciferase reporter assay were used to analyze markers of inflammation cytokines and podocyte injury. Results. miRNA-155 was found to be highly expressed in serum and kidney tissue of mice with diabetic nephropathy and in cultured podocytes, accompanied by elevated levels of inflammatory factors. Inhibition of miRNA-155 can reduce proteinuria and ACR levels, diminish the secretion of inflammatory molecules, improve kidney function, inhibit podocyte foot fusion, and reverse renal pathological changes in diabetic nephropathy mice. Overexpression of miRNA-155 in vitro can increase inflammatory molecule production in podocytes and aggravates podocyte injury, while miRNA-155 inhibition suppresses inflammatory molecule production in podocytes and reduces podocyte injury. A luciferase assay confirmed that miRNA-155 could selectively bind to 3 ′ -UTR of SIRT1, resulting in decreased SIRT1 expression. In addition, SIRT1 siRNA could offset SIRT1 upregulation and enhance inflammatory factor secretion in podocytes, induced by the miRNA-155 inhibitor. Conclusions. These findings strongly support the hypothesis that miRNA-155 inhibits podocyte inflammation and reduces podocyte injury through SIRT1 silencing. miRNA-155 suppression therapy may be useful for the management of diabetic nephropathy.

scholarly journals The High Expression of PTPRH Is Associated with Poor Prognosis of Human Lung Adenocarcinoma

2021 ◽  
Vol 2021 ◽  
pp. 1-9
Author(s):  
Aifeng Chen ◽  
Shibiao Ding ◽  
Xiaoqiang Shen ◽  
Xuai Lin
Keyword(s):  
Lung Adenocarcinoma ◽  
Roc Curve ◽  
Immunohistochemical Staining ◽  
Poor Prognosis ◽  
Clinicopathological Characteristics ◽  
The Cancer Genome Atlas ◽  
Disease Stage ◽  
Tissue Samples ◽  
Qrt Pcr ◽  
Kaplan Meier

Objective. The aim of the study is to explore the prognosis value of PTPRH in patients with lung adenocarcinoma (LUAD). Methods. Oncomine, UALCAN, and GEPIA databases were employed to examine the differential expression of PTPRH between LUAD and adjacent tissues. 100 pairs of LUAD and adjacent tissue samples were involved in this study. qRT-PCR and immunohistochemical staining were performed. Meanwhile, we analyzed The Cancer Genome Atlas (TCGA) data to investigate the correlation between PTPRH gene expression and clinicopathological characteristics. Kaplan-Meier analysis and univariate and multivariate Cox analyses were performed to estimate the relationship between PTPRH expression and LUAD prognosis. The evaluation performance was verified by drawing a ROC curve. In addition, through GSEA, the changes of PTPRH expression were analyzed by GSEA to screen out primarily affected signaling pathway. Results. Oncomine, UALCAN, and GEPIA databases showed that the mRNA expression of PTPRH in LUAD tissues was significantly higher than that in adjacent tissues. qRT-PCR and immunohistochemical staining indicated the mRNA and protein levels of PTPRH in LUAD tissues were markedly upregulated. TCGA data showed that the expression of PTPRH was significantly correlated with T stage and disease stage. Kaplan-Meier analysis showed that the patients with high PTPRH expression had a poor prognosis. Univariate and multivariate Cox analyses exhibited that PTPRH expression could act as an independent prognostic factor for LUAD. The ROC curve showed that PTPRH combined with various clinicopathological features could effectively predict the prognosis of LUAD. Finally, GSEA indicated that changes in PTPRH expression level may affect p53, VEGF, Notch, and mTOR cancer-related signaling pathways. Conclusion. Our results demonstrated that PTPRH was highly expressed in LUAD and may be closely correlated with the poor prognosis of LUAD patients.

scholarly journals Genetics and Epigenetics of Diabetic Nephropathy

2015 ◽  
Vol 1 (1) ◽  
pp. 42-51 ◽  
Author(s):  
Ruijie Liu ◽  
Kyung Lee ◽  
John Cijiang He
Keyword(s):  
Diabetic Nephropathy ◽  
Drug Targets ◽  
Rare Variants ◽  
Rapid Development ◽  
Strong Association ◽  
Kidney Tissue ◽  
Epigenetic Modifications ◽  
Diabetic Patients ◽  
Epigenetic Changes ◽  
Tissue Samples

Background: Diabetic nephropathy (DN) is the most common cause of end-stage renal disease (ESRD) in the USA and worldwide, contributing to significant morbidity and mortality in diabetic patients. A genetic factor for the development of DN is strongly implicated, as only one third of diabetic patients eventually develop kidney disease. Growing evidence also supports an important role of epigenetic modifications in DN. Summary: Multiple studies have been performed to identify risk genes and loci associated with DN. So far, only several genes and loci have been identified, none of which showed a strong association with DN. Therefore, a better study design with a larger sample size to identify rare variants and a clinically defined patient population to identify genes and loci associated with progressive DN are still needed. In addition to genetic factors, epigenetic modifications, such as DNA methylation, histone modifications and microRNAs, also play a major role in the pathogenesis of DN through a second layer of gene regulation. Although a major progress has been made in this field, epigenetic studies in DN are still in the early phase and have been limited mostly due to the heterogeneity of kidney tissue samples with multiple cells. However, rapid development of high-throughput genome-wide techniques will help us to better identify genetic variants and epigenetic changes in DN. Key Message: Understanding of genetic and epigenetic changes in DN is needed for the development of new biomarkers and better drug targets against DN. Summarized in this review are important recent findings on genetic and epigenetic studies in the field of DN.

Microprobe analysis of inclusion bodies related to two benzofuran derivatives

1987 ◽  
Vol 45 ◽  
pp. 672-673
Author(s):  
T. L. Benning ◽  
P. Ingram ◽  
J. D. Shelburne
Keyword(s):  
Inclusion Bodies ◽  
Uranyl Acetate ◽  
Multiple Organ ◽  
High Dose ◽  
En Bloc ◽  
Tissue Samples ◽  
Transmission Electron ◽  
Organ Systems ◽  
Dense Inclusion ◽  
Melanin Complex

Two benzofuran derivatives, chlorpromazine and amiodarone, are known to produce inclusion bodies in human tissues. Prolonged high dose chlorpromazine therapy causes hyperpigmentation of the skin with electron-dense inclusion bodies present in dermal histiocytes and endothelial cells ultrastructurally. The nature of the deposits is not known although a drug-melanin complex has been hypothesized. Amiodarone may also cause cutaneous hyperpigmentation and lamellar lysosomal inclusion bodies have been demonstrated within the cells of multiple organ systems. These lamellar bodies are believed to be the product of an amiodarone-induced phospholipid storage disorder. We performed transmission electron microscopy (TEM) and energy dispersive x-ray microanalysis (EDXA) on tissue samples from patients treated with these drugs, attempting to detect the sulfur atom of chlorpromazine and the iodine atom of amiodarone within their respective inclusion bodies.A skin biopsy from a patient with hyperpigmentation due to prolonged chlorpromazine therapy was fixed in 4% glutaraldehyde and processed without osmium tetroxide or en bloc uranyl acetate for Epon embedding.

Hepatocyte ultrastructural alterations in perimetastatic areas

1996 ◽  
Vol 54 ◽  
pp. 768-769
Author(s):  
H. J. Finol ◽  
M. E. Correa ◽  
L.A. Sosa ◽  
A. Márquez ◽  
N.L. Díaz
Keyword(s):  
Malignant Tumor ◽  
Malignant Tumors ◽  
Surrounding Tissue ◽  
Ultrastructural Changes ◽  
Pancreas Carcinoma ◽  
Duct Carcinoma ◽  
Tissue Samples ◽  
Primary Malignant Tumor ◽  
Transmission Electron ◽  
Remote Sites

In classical oncological literature two mechanisms for tissue aggression in patients with cancer have been described. The first is the progressive invasion, infiltration and destruction of tissues surrounding primary malignant tumor or their metastases; the other includes alterations produced in remote sites that are not directly affected by any focus of disease, the so called paraneoplastic phenomenon. The non-invaded tissue which surrounds a primary malignant tumor or its metastases has been usually considered a normal tissue . In this work we describe the ultrastructural changes observed in hepatocytes located next to metastases from diverse malignant tumors.Hepatic biopsies were obtained surgically in patients with different malignant tumors which metatastized in liver. Biopsies included tumor mass, the zone of macroscopic contact between the tumor and the surrounding tissue, and the tissue adjacent to the tumor but outside the macroscopic area of infiltration. The patients (n = 5), 36–75 years old, presented different tumors including rhabdomyosarcoma, leiomyosarcoma, pancreas carcinoma, biliar duct carcinoma and colon carcinoma. Tissue samples were processed with routine techniques for transmission electron microscopy and observed in a Hitachi H-500 electron microscope.

scholarly journals Evidence for the Use of Mucus Swabs to Detect Renibacterium salmoninarum in Brook Trout

Pathogens ◽  
2021 ◽  
Vol 10 (4) ◽  
pp. 460
Author(s):  
Tawni B. Riepe ◽  
Victoria Vincent ◽  
Vicki Milano ◽  
Eric R. Fetherman ◽  
Dana L. Winkelman
Keyword(s):  
Brook Trout ◽  
Sampling Methods ◽  
Kidney Tissue ◽  
Detection Methods ◽  
Fish Health ◽  
Wild Populations ◽  
Renibacterium Salmoninarum ◽  
Tissue Samples ◽  
Future Studies ◽  
Aquaculture Facilities

Efforts to advance fish health diagnostics have been highlighted in many studies to improve the detection of pathogens in aquaculture facilities and wild fish populations. Typically, the detection of a pathogen has required sacrificing fish; however, many hatcheries have valuable and sometimes irreplaceable broodstocks, and lethal sampling is undesirable. Therefore, the development of non-lethal detection methods is a high priority. The goal of our study was to compare non-lethal sampling methods with standardized lethal kidney tissue sampling that is used to detect Renibacterium salmoninarum infections in salmonids. We collected anal, buccal, and mucus swabs (non-lethal qPCR) and kidney tissue samples (lethal DFAT) from 72 adult brook trout (Salvelinus fontinalis) reared at the Colorado Parks and Wildlife Pitkin Brood Unit and tested each sample to assess R. salmoninarum infections. Standard kidney tissue detected R. salmoninarum 1.59 times more often than mucus swabs, compared to 10.43 and 13.16 times more often than buccal or anal swabs, respectively, indicating mucus swabs were the most effective and may be a useful non-lethal method. Our study highlights the potential of non-lethal mucus swabs to sample for R. salmoninarum and suggests future studies are needed to refine this technique for use in aquaculture facilities and wild populations of inland salmonids.

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