Quantification of gene expression in urinary sediment for the study of renal diseases

Background. The association between hypothyroidism and renal diseases has been described in many studies. Nigella Sativa was among the recently reported natural product that has the potential to prevent renal tissue damage and fibrosis. The aim of this study was to evaluate the possible protective effect of thymoquinone on the structure of the renal cortex of hypothyroid rats and explore the mechanism behind it. Methods. An experimental model of hypothyroidism was induced in adult male Wistar rats by administration of propylthiouracil (6 mg/kg/body weight). One hypothyroid group was treated with thymoquinone at the dose of 50 mg/kg/body weight and compared to the untreated group. Thyroid function and oxidant/antioxidant status were assessed in the serum. Catalase gene expression was assessed using the real-time polymerase chain reaction. The kidney was assessed both histologically and immunohistochemically. Results. Administration of propylthiouracil resulted in a significant decrease in the serum levels of nitric oxide, reduced glutathione, and superoxide dismutase activity while the level of malondialdehyde significantly (p<0.001) increased. Administration of thymoquinone alleviated this effect on the thyroid hormones and significantly increased the serum levels of antioxidants. Thymoquinone significantly (p<0.001) upregulated catalase transcription by about 24-fold and could block the hypothyroidism-induced glomerular and tubular injury. Conclusion. Thymoquinone may have a potential protective effect against hypothyroidism-induced renal injury acting through the attenuation of the oxidative stress and upregulation of renal catalase gene expression.

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Melittin Inhibits TGF-β-Induced Pro-Fibrotic Gene Expression Through the Suppression of the TGFβRII-Smad, ERK1/2 and JNK-Mediated Signaling Pathway

The American Journal of Chinese Medicine ◽

10.1142/s0192415x14500712 ◽

2014 ◽

Vol 42 (05) ◽

pp. 1139-1152 ◽

Cited By ~ 13

Author(s):

Su-Hyun Park ◽

Hyun-Ji Cho ◽

Yun-Jeong Jeong ◽

Jae-Moon Shin ◽

Jeong-Han Kang ◽

...

Keyword(s):

Gene Expression ◽

Type I Collagen ◽

Bee Venom ◽

Renal Diseases ◽

Type I ◽

Inhibitory Effects ◽

Genes Expression ◽

Kidney Fibroblast ◽

Pai 1 ◽

Excessive Accumulation

Renal fibrosis is characterized by the excessive accumulation of extracellular matrix (ECM) proteins such as type I collagen, fibronectin, and by the increased expression of PAI-1. This study evaluated the anti-fibrotic effect of bee venom and its major compounds (melittin and apamin) on TGF-β-induced pro-fibrotic gene expression. Bee venom and melittin significantly suppressed type I collagen, fibronectin, and PAI-1 protein expression in the TGF-β-treated kidney fibroblast. However, apamin only inhibited the expression of fibronectin and type I collagen. These results indicated that the inhibitory effects of bee venom on TGF-β-induced pro-fibrotic gene expression are caused by melittin. Moreover, we attempted to elucidate mechanisms underlying the anti-fibrotic effect of melittin. Melittin dramatically inhibited the phosphorylation of TGFβRII and Smad2/3. Also, melittin inhibited the phosphorylation of ERK1/2 and JNK, but not the phosphorylation of PI3K, Akt, and p38. These results suggested that melittin inhibits TGF-β-induced pro-fibrotic genes expression through the suppression of TGFβR-Smad2/3, ERK1/2, and JNK phosphorylation, and melittin can be used as a clinical drug for the treatment of fibrosis associated with renal diseases.

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Inflammatory cytokine gene expression in the urinary sediment of patients with lupus nephritis

Arthritis & Rheumatism ◽

10.1002/art.11062 ◽

2003 ◽

Vol 48 (5) ◽

pp. 1326-1331 ◽

Cited By ~ 55

Author(s):

Rebecca Wing-Yan Chan ◽

Lai-Shan Tam ◽

Edmund Kwok-Ming Li ◽

Fernand Mac-Moune Lai ◽

Kai-Ming Chow ◽

...

Keyword(s):

Gene Expression ◽

Lupus Nephritis ◽

Inflammatory Cytokine ◽

Cytokine Gene Expression ◽

Cytokine Gene ◽

Urinary Sediment

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The urine biomarker panel [IGFBP7]x[TIMP-2] (NephroCheck® parameter) does not correlate with IGFBP7 and TIMP-2 gene expression in urinary sediment

PLoS ONE ◽

10.1371/journal.pone.0188316 ◽

2017 ◽

Vol 12 (11) ◽

pp. e0188316 ◽

Cited By ~ 2

Author(s):

Daniela Knafl ◽

Markus Müller ◽

Sahra Pajenda ◽

Zeynep Genc ◽

Manfred Hecking ◽

...

Keyword(s):

Gene Expression ◽

Urinary Sediment ◽

Biomarker Panel ◽

Urine Biomarker

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Histone Lysine Methylation in TGF-β1 Mediated p21 Gene Expression in Rat Mesangial Cells

BioMed Research International ◽

10.1155/2016/6927234 ◽

2016 ◽

Vol 2016 ◽

pp. 1-9 ◽

Cited By ~ 4

Author(s):

Qiaoyan Guo ◽

Xiaoxia Li ◽

Hongbo Han ◽

Chaoyuan Li ◽

Shujun Liu ◽

...

Keyword(s):

Gene Expression ◽

Transforming Growth Factor ◽

Mesangial Cells ◽

Gene Promoter ◽

Renal Diseases ◽

Lysine Methylation ◽

Histone Lysine Methylation ◽

Histone Lysine ◽

Chronic Renal Diseases

Transforming growth factor beta1- (TGF-β1-) induced p21-dependent mesangial cell (MC) hypertrophy plays a key role in the pathogenesis of chronic renal diseases including diabetic nephropathy (DN). Increasing evidence demonstrated the role of posttranscriptional modifications (PTMs) in the event; however, the precise regulatory mechanism of histone lysine methylation remains largely unknown. Here, we examined the roles of both histone H3 lysine 4 and lysine 9 methylations (H3K4me/H3K9me) in TGF-β1 induced p21 gene expression in rat mesangial cells (RMCs). Our results indicated that TGF-β1 upregulated the expression of p21 gene in RMCs, which was positively correlated with the increased chromatin marks associated with active genes (H3K4me1/H3K4me2/H3K4me3) and negatively correlated with the decreased levels of repressive marks (H3K9me2/H3K9me3) at p21 gene promoter. TGF-β1 also elevated the recruitment of the H3K4 methyltransferase (HMT) SET7/9 to the p21 gene promoter. SET7/9 gene silencing with small interfering RNAs (siRNAs) significantly abolished the TGF-β1 induced p21 gene expression. Taken together, these results reveal the key role of histone H3Kme in TGF-β1 mediated p21 gene expression in RMC, partly through HMT SET7/9 occupancy, suggesting H3Kme and SET7/9 may be potential renoprotective agents in managing chronic renal diseases.

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The Non-coding MicroRNA-223 is a Promising Biomarker of Chronic Kidney Disease

EMJ Nephrology ◽

10.33590/emjnephrol/21-00057 ◽

2021 ◽

pp. 91-95

Author(s):

Valerie Metzinger-Le Meuth ◽

Laurent Metzinger

Keyword(s):

Gene Expression ◽

Chronic Kidney Disease ◽

Kidney Disease ◽

Renal Diseases ◽

Mirna Regulation ◽

Mrna Targets ◽

Cardiovascular Damage ◽

Non Coding Rnas ◽

Post Transcriptional Regulation

Renal diseases are consecutive to a deregulation of gene expression regulated by non-coding RNAs. These non-coding RNAs were discovered at the turn of the 21st century when it was established that post-transcriptional regulation was performed through small non-coding RNAs, known as microRNAs (miRNAs). Up to 3,000 miRNAs are expressed by human cells. They are small, single-stranded nucleic acids, which trigger translational repression of mRNA by base-pairing with the 3′ untranslated region of their mRNA targets. In addition to miRNA regulation, it was also demonstrated that 60,000 long non-coding RNAs are expressed in the human cell and that they are able to regulate gene expression at all levels. The roles of these various RNA families are just beginning to be understood in the field of nephrology. In the past decade, the authors and various others have published that several miRNAs are deregulated during the onset of chronic kidney disease (CKD) and are associated with cardiovascular damage. This review focuses on miRNA-223 (miR-223) as its expression is increased in vivo in the large vessels of a mouse model of CKD, whereas it is diminished in the serum of both mice and human patients with CKD. In patients, miR-223 expression was correlated with all-cause mortality, as well as cardiovascular and renal events. Molecular clues were given by a multi-omics approach, indicating that miR-223 modulates gene regulation at all levels including mRNA expression, protein amounts, and metabolic molecule accumulation. miR-223 is thus a potential target to prevent or treat complications of CKD pathogenesis.

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Abstract 437: Endothelial Cell Gene Expression in Hypertensive Mice

Arteriosclerosis Thrombosis and Vascular Biology ◽

10.1161/atvb.37.suppl_1.437 ◽

2017 ◽

Vol 37 (suppl_1) ◽

Author(s):

Stephen J Wilson ◽

Kate L Lowe ◽

Michael P Bokoch ◽

Yoga Srinivasan ◽

Roland S Wu ◽

...

Keyword(s):

Gene Expression ◽

Endothelial Cells ◽

Angiotensin Ii ◽

Weight Ratio ◽

Heart Tissue ◽

Renal Diseases ◽

Fatty Acid Transport ◽

Premature Cardiovascular Disease ◽

Matrix Metabolism ◽

Endothelial Genes

Hypertension is the major risk factor for premature cardiovascular disease, morbidity and mortality. It is the leading cause of death and disability globally, contributing to cardiovascular and renal diseases, including stroke, heart failure, coronary heart disease, and chronic kidney disease. We sought to determine the transcriptional changes that occur in cardiac endothelial cells secondary to hypertension, using ribosomal profiling of mice expressing the ribosomal protein Rpl22 tagged with the hemagglutinin (HA) epitope under control of Tie2-Cre (Tie2-Cre:RiboTag mice). Immunoprecipitation of HA-Rpl22 from heart tissue isolated from Tie2-Cre:RiboTag mice resulted in an approximate 10-fold enrichment of endothelial genes (Pecam-1, VE-cadherin) relative to genes highly expressed in cardiomyocytes (Tnnt2, Fabp3) as assessed by RT-PCR. Subsequently, Tie2-Cre:RiboTag mice were implanted with osmotic pumps delivering saline or angiotensin II (1000ng/kg/min) for 4 weeks. Angiotensin II infusion significantly increased blood pressure (p<0.0001, n=7-8) resulting in an increased heart to body weight ratio (6.07 ± 0.48 vs 8.14 ± 0.73; p<0.05, n=7-8). Transcriptomes from Tie2-Cre positive cells within the heart were isolated using Ribotag ribosomal immunoprecipitation and RNA-seq was performed. Five hundred and eighty seven differentially expressed genes were detected (fold change >1.5, p<0.05, n=3), representing pathways such as extracellular matrix metabolism, angiogenesis and fatty acid transport. Analysis of hypertension associated gene expression in endothelial cells across different organs may reveal the mechanism of hypertension-associated diseases.

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FC 020SINGLE CELL SEQUENCING REVEALS TRANSCRIPTIONAL SIGNATURES AND CELL CROSSTALK IN PATIENTS WITH HYPERTENSIVE NEPHROPATHY

Nephrology Dialysis Transplantation ◽

10.1093/ndt/gfab132.003 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

Rong Tang ◽

Ting Meng ◽

Wei Lin ◽

Yong Zhong

Keyword(s):

Gene Expression ◽

Renal Diseases ◽

Receptor Interaction ◽

Pathway Enrichment Analysis ◽

Specific Gene ◽

Hypertensive Nephropathy ◽

Specific Gene Expression ◽

Distinct Cell ◽

Fibrotic Process ◽

Cell Cell

Abstract Background and Aims Hypertensive nephropathy (HTN) is one of the leading causes of end-stage renal disease (ESRD). HTN is characterized by injury to the glomerulus, arterioles and tubulointerstitium, yet the precise mechanisms and cell-specific gene expression changes are still unknown. This study used single-cell RNA sequencing (scRNA-seq) to explore novel molecular mechanisms and the gene targets for HTN. Method The gene expression profiles of human renal biospsy samples obtained from subjects with HTN and pre-transplant healthy living controls were determined by scRNA-seq technology. Then the differentially expressed genes (DEGs) and their functions were identified. Gene ontology (GO) and the Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis were conducted, and the ligand-receptor interaction among different cell populations were anlyzed. Results 18 distinct cell clusters were identified in kidney from HTN patients. Endothelial cells overexpressed LRG1, a pleiotropic factor linked to apoptosis and inflammation, which was validated at proteome level in kidney from HTN, providing a potential novel molecular target. In HTN patients, mesangial cells highly expressed proliferation related signatures (MGST1, TMSB10, EPS8 and IER2) not detected in renal diseases before. The upregualted genes in tubules of HTN were mainly participating in inflammatory signatures including IFN-γ signature, IL-17 signaling and TLR signaling. Specific gene expression of kidney-resident immune cells including dendritic cells and CD8+ T cells revealed abnormal regulation associated with cell adhesion and inflammation. Furthermore, the receptor-ligand interactions analysis indicated cell-cell crosstalks in kidney contribute to recruitment and infiltration of inflammatory cells into kidneys, and fibrotic process in hypertensive renal injury. Conclusion In brief, our data identifies distinct cell-specific gene expression profile, pathogenic signaling pathways and potential cell-cell communication in pathogenesis of HTN. These findings will provide a promising novel landscape for mechanisms and treatment of HTN.

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Gremlin activates the Notch pathway linked to renal inflammation

Clinical Science ◽

10.1042/cs20171553 ◽

2018 ◽

Vol 132 (11) ◽

pp. 1097-1115 ◽

Cited By ~ 9

Author(s):

Carolina Lavoz ◽

Jonay Poveda ◽

Laura Marquez-Exposito ◽

Sandra Rayego-Mateos ◽

Raul R. Rodrigues-Diez ◽

...

Keyword(s):

Gene Expression ◽

Kidney Development ◽

Nuclear Translocation ◽

Kidney Diseases ◽

Notch Pathway ◽

Renal Diseases ◽

Notch Inhibition ◽

Anti Inflammatory ◽

Regulated Gene Expression

Preclinical studies suggest that Gremlin participates in renal damage and could be a potential therapeutic target for human chronic kidney diseases. Inflammation is a common characteristic of progressive renal disease, and therefore novel anti-inflammatory therapeutic targets should be investigated. The Notch signaling pathway is involved in kidney development and is activated in human chronic kidney disease, but whether Gremlin regulates the Notch pathway has not been investigated. In cultured tubular cells, Gremlin up-regulated gene expression of several Notch pathway components, increased the production of the canonical ligand Jagged-1, and caused the nuclear translocation of active Notch-1 (N1ICD). In vivo administration of Gremlin into murine kidneys elicited Jagged-1 production, increased N1ICD nuclear levels, and up-regulated the gene expression of the Notch effectors hes-1 and hey-1. All these data clearly demonstrate that Gremlin activates the Notch pathway in the kidney. Notch inhibition using the γ-secretase inhibitor DAPT impaired renal inflammatory cell infiltration and proinflammatory cytokines overexpression in Gremlin-injected mice and in experimental models of renal injury. Moreover, Notch inhibition blocked Gremlin-induced activation of the canonical and noncanonical nuclear factor-κB (NF-κB) pathway, identifying an important mechanism involved in the anti-inflammatory actions of Notch inhibition. In conclusion, Gremlin activates the Notch pathway in the kidney and this is linked to NF-κB-mediated inflammation, supporting the hypothesis that Notch inhibition could be a potential anti-inflammatory strategy for renal diseases.

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