Acetaminophen is a drug used to treat many conditions as headache, muscle aches, arthritis, backache, toothache, and fever between others, but collateral effects of this drug are not well known yet. Here is tested its...Acetaminophen is a drug used to treat many conditions as headache, muscle aches, arthritis, backache, toothache, and fever between others, but collateral effects of this drug are not well known yet. Here is tested its effect on proximal tubule epithelium. Acetaminophen (APAP) at doses of 200, 500, 1000 and 1500 mg/Kg i.p. caused cell damage and changes in F-actin distribution in the proximal tubule of male Wistar rats. After 48 hours of treatment, the proximal tubule epithelium showed tumefaction and necrosis. Dose of 200 mg/kg decreased the F-actin and was observed a structure in patches in the basal cytoplasm of epithelial cells of the proximal tubule. This effect was increased depending on the administered dose. Dose of 1000 mg/kg produced the highest histological damage and changes in the actin cytoskeleton. Results of this study suggested that nephrotoxic damage produced by high doses of APAP included breakdown of cytoskeleton in proximal tubule epithelium.展开更多
The mechanism of how SARS-CoV-2 causes severe multi-organ failure is largely unknown. Acute kidney injury(AKI) is one of the frequent organ damage in severe COVID-19 patients. Previous studies have shown that human re...The mechanism of how SARS-CoV-2 causes severe multi-organ failure is largely unknown. Acute kidney injury(AKI) is one of the frequent organ damage in severe COVID-19 patients. Previous studies have shown that human renal tubule cells could be the potential host cells targeted by SARS-CoV-2. Traditional cancer cell lines or immortalized cell lines are genetically and phenotypically different from host cells. Animal models are widely used, but often fail to reflect a physiological and pathogenic status because of species tropisms. There is an unmet need for normal human epithelial cells for disease modeling. In this study, we successfully established long term cultures of normal human kidney proximal tubule epithelial cells(KPTECs) in 2 D and 3 D culture systems using conditional reprogramming(CR) and organoids techniques.These cells had the ability to differentiate and repair DNA damage, and showed no transforming property. Importantly, the CR KPTECs maintained lineage function with expression of specific transporters(SLC34 A3 and cubilin). They also expressed angiotensin-converting enzyme 2(ACE2), a receptor for SARS-CoV and SARS-CoV-2. In contrast, cancer cell line did not express endogenous SLC34 A3, cubilin and ACE2. Very interestingly, ACE2 expression was around twofold higher in 3 D organoids culture compared to that in 2 D CR culture condition. Pseudovirion assays demonstrated that SARS-CoV spike(S) protein was able to enter CR cells with luciferase reporter. This integrated 2 D CR and 3 D organoid cultures provide a physiological ex vivo model to study kidney functions, innate immune response of kidney cells to viruses, and a novel platform for drug discovery and safety evaluation.展开更多
Background:Ischemic acute kidney injury(AKI)is a common syndrome associated with considerable mortality and healthcare costs.Up to now,the underlying pathogenesis of ischemic AKI remains incompletely understood,and sp...Background:Ischemic acute kidney injury(AKI)is a common syndrome associated with considerable mortality and healthcare costs.Up to now,the underlying pathogenesis of ischemic AKI remains incompletely understood,and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking.Here,this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing(scRNA-seq)analysis in kidneys.Methods:In this study,scRNA-seq technology was applied to kidneys from two ischemic AKI patients,and three human public scRNA-seq datasets were collected as controls.Differentially expressed genes(DEGs)and cell clusters of kidneys were determined.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,as well as the ligand-receptor interaction between cells,were performed.We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion(I/R)injury induced AKI mice through immunohistochemistry staining.Results:15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control.The injured proximal tubules(PT)displayed a proapoptotic and proinflammatory phenotype.PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47,RASSF4,EBAG9,IER3,SASH1,SEPTIN7,and NUB1,which have not been reported in ischemic AKI previously.Several hub genes were validated in kidneys from human AKI and renal I/R injury mice,respectively.Furthermore,PT highly expressed DEGs enriched in endoplasmic reticulum stress,autophagy,and retinoic acid-inducible gene I(RIG-I)signaling.DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain(NOD)-like receptor signaling,estrogen signaling,interleukin(IL)-12 signaling,and IL-17 signaling.Overexpressed genes in kidney-resident immune cells including macrophages,natural killer T(NKT)cells,monocytes,and dendritic cells were associated with leukocyte activation,chemotaxis,cell adhesion,and complement activation.In addition,the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI.Conclusion:Together,this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients,altered signaling pathways,and potential cell-cell crosstalk in the development of AKI.These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.展开更多
Background Fanconi-Debré-de Toni syndrome(also known as Fanconi renotubular syndrome,or FRST)profoundly increased the understanding of the functions of the proximal convoluted tubule(PCT)and provided important in...Background Fanconi-Debré-de Toni syndrome(also known as Fanconi renotubular syndrome,or FRST)profoundly increased the understanding of the functions of the proximal convoluted tubule(PCT)and provided important insights into the pathophysiology of several kidney diseases and drug toxicities.Data sources We searched Pubmed and Scopus databases to find relevant articles about FRST.This review article focuses on the physiology of the PCT,as well as on the physiopathology of FRST in children,its diagnosis,and treatment.Results FRST encompasses a wide variety of inherited and acquired PCT alterations that lead to impairment of PCT reab-sorption.In children,FRST often presents as a secondary feature of systemic disorders that impair energy supply,such as Lowe's syndrome,Dent's disease,cystinosis,hereditary fructose intolerance,galactosemia,tyrosinemia,Alport syndrome,and Wilson's disease.Although rare,congenital causes of FRST greatly impact the morbidity and mortality of patients and impose diagnostic challenges.Furthermore,its treatment is diverse and considers the ability of the clinician to identify the correct etiology of the disease.Conclusion The early diagnosis and treatment of pediatric patients with FRST improve the prognosis and the quality of life.展开更多
文摘Acetaminophen is a drug used to treat many conditions as headache, muscle aches, arthritis, backache, toothache, and fever between others, but collateral effects of this drug are not well known yet. Here is tested its effect on proximal tubule epithelium. Acetaminophen (APAP) at doses of 200, 500, 1000 and 1500 mg/Kg i.p. caused cell damage and changes in F-actin distribution in the proximal tubule of male Wistar rats. After 48 hours of treatment, the proximal tubule epithelium showed tumefaction and necrosis. Dose of 200 mg/kg decreased the F-actin and was observed a structure in patches in the basal cytoplasm of epithelial cells of the proximal tubule. This effect was increased depending on the administered dose. Dose of 1000 mg/kg produced the highest histological damage and changes in the actin cytoskeleton. Results of this study suggested that nephrotoxic damage produced by high doses of APAP included breakdown of cytoskeleton in proximal tubule epithelium.
基金supported by the National Natural Science Foundation of China (81571396 and 81771528)Science, Technology and Innovation Commission of Shenzhen Municipality (JCYJ20170411090932146, JCYJ20170818110544730)。
文摘The mechanism of how SARS-CoV-2 causes severe multi-organ failure is largely unknown. Acute kidney injury(AKI) is one of the frequent organ damage in severe COVID-19 patients. Previous studies have shown that human renal tubule cells could be the potential host cells targeted by SARS-CoV-2. Traditional cancer cell lines or immortalized cell lines are genetically and phenotypically different from host cells. Animal models are widely used, but often fail to reflect a physiological and pathogenic status because of species tropisms. There is an unmet need for normal human epithelial cells for disease modeling. In this study, we successfully established long term cultures of normal human kidney proximal tubule epithelial cells(KPTECs) in 2 D and 3 D culture systems using conditional reprogramming(CR) and organoids techniques.These cells had the ability to differentiate and repair DNA damage, and showed no transforming property. Importantly, the CR KPTECs maintained lineage function with expression of specific transporters(SLC34 A3 and cubilin). They also expressed angiotensin-converting enzyme 2(ACE2), a receptor for SARS-CoV and SARS-CoV-2. In contrast, cancer cell line did not express endogenous SLC34 A3, cubilin and ACE2. Very interestingly, ACE2 expression was around twofold higher in 3 D organoids culture compared to that in 2 D CR culture condition. Pseudovirion assays demonstrated that SARS-CoV spike(S) protein was able to enter CR cells with luciferase reporter. This integrated 2 D CR and 3 D organoid cultures provide a physiological ex vivo model to study kidney functions, innate immune response of kidney cells to viruses, and a novel platform for drug discovery and safety evaluation.
基金National Key Research and Development Program of China(No.2020YFC2005000)Key Research and Development Program of Hunan province(No.2020WK2008)+3 种基金science and technology innovation Program of Hunan Province(No.2020RC5002)Natural Science Foundation of Hunan Province(Nos.2022JJ30070,2021JJ31130 and 2021JJ31057)Project of Health Commission of Hunan Province(Nos.A202303050036 and 202104101009)"Yiluqihang Shenmingyuanyang"medical development and Scientific Research Fund project on Kidney Diseases(No.SMYY20220301001)
文摘Background:Ischemic acute kidney injury(AKI)is a common syndrome associated with considerable mortality and healthcare costs.Up to now,the underlying pathogenesis of ischemic AKI remains incompletely understood,and specific strategies for early diagnosis and treatment of ischemic AKI are still lacking.Here,this study aimed to define the transcriptomic landscape of AKI patients through single-cell RNA sequencing(scRNA-seq)analysis in kidneys.Methods:In this study,scRNA-seq technology was applied to kidneys from two ischemic AKI patients,and three human public scRNA-seq datasets were collected as controls.Differentially expressed genes(DEGs)and cell clusters of kidneys were determined.Gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway enrichment analysis,as well as the ligand-receptor interaction between cells,were performed.We also validated several DEGs expression in kidneys from human ischemic AKI and ischemia/reperfusion(I/R)injury induced AKI mice through immunohistochemistry staining.Results:15 distinct cell clusters were determined in kidney from subjects of ischemic AKI and control.The injured proximal tubules(PT)displayed a proapoptotic and proinflammatory phenotype.PT cells of ischemic AKI had up-regulation of novel pro-apoptotic genes including USP47,RASSF4,EBAG9,IER3,SASH1,SEPTIN7,and NUB1,which have not been reported in ischemic AKI previously.Several hub genes were validated in kidneys from human AKI and renal I/R injury mice,respectively.Furthermore,PT highly expressed DEGs enriched in endoplasmic reticulum stress,autophagy,and retinoic acid-inducible gene I(RIG-I)signaling.DEGs overexpressed in other tubular cells were primarily enriched in nucleotide-binding and oligomerization domain(NOD)-like receptor signaling,estrogen signaling,interleukin(IL)-12 signaling,and IL-17 signaling.Overexpressed genes in kidney-resident immune cells including macrophages,natural killer T(NKT)cells,monocytes,and dendritic cells were associated with leukocyte activation,chemotaxis,cell adhesion,and complement activation.In addition,the ligand-receptor interactions analysis revealed prominent communications between macrophages and monocytes with other cells in the process of ischemic AKI.Conclusion:Together,this study reveals distinct cell-specific transcriptomic atlas of kidney in ischemic AKI patients,altered signaling pathways,and potential cell-cell crosstalk in the development of AKI.These data reveal new insights into the pathogenesis and potential therapeutic strategies in ischemic AKI.
文摘Background Fanconi-Debré-de Toni syndrome(also known as Fanconi renotubular syndrome,or FRST)profoundly increased the understanding of the functions of the proximal convoluted tubule(PCT)and provided important insights into the pathophysiology of several kidney diseases and drug toxicities.Data sources We searched Pubmed and Scopus databases to find relevant articles about FRST.This review article focuses on the physiology of the PCT,as well as on the physiopathology of FRST in children,its diagnosis,and treatment.Results FRST encompasses a wide variety of inherited and acquired PCT alterations that lead to impairment of PCT reab-sorption.In children,FRST often presents as a secondary feature of systemic disorders that impair energy supply,such as Lowe's syndrome,Dent's disease,cystinosis,hereditary fructose intolerance,galactosemia,tyrosinemia,Alport syndrome,and Wilson's disease.Although rare,congenital causes of FRST greatly impact the morbidity and mortality of patients and impose diagnostic challenges.Furthermore,its treatment is diverse and considers the ability of the clinician to identify the correct etiology of the disease.Conclusion The early diagnosis and treatment of pediatric patients with FRST improve the prognosis and the quality of life.
