Silencing of USP22 suppresses high glucose-induced apoptosis, ROS production and inflammation in podocytes

Diabetic nephropathy is the complication of diabetes mellitus that can lead to chronic renal failure. Reactive oxygen species (ROS) production plays an important role in its pathological process. Previous studies showed that carnosine may reduce diabetic nephropathy by antioxidant effect. However, the molecular mechanism of its antioxidant was not fully understood. In the current study, we developed high glucose containing different concentrations of carnosine to reduce ROS levels and podocytes apoptosis, and Cell Counting Kit-8 test was used to observe the cell viability. Carnosine (5-20mM) was found to protect mouse podocytes (MPC5) cells from HG-induced injury. Quantitative real-time PCR, Western blotting, and immunofluorescence staining revealed that high glucose induced ROS levels and podocytes apoptosis were downregulated by PI3K/AKT and Nrf2 signaling pathways. The current findings suggest that carnosine may reduce ROS levels and MPC5 cells apoptosis by PI3K/AKT and Nrf2 signaling pathways activation.

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High-glucose-induced apoptosis, ROS production and pro-inflammatory response in cardiomyocytes is attenuated by metformin treatment via PP2A activation

Journal of Biosciences ◽

10.1007/s12038-020-00096-5 ◽

2020 ◽

Vol 45 (1) ◽

Author(s):

Gang Cheng ◽

Lihuan Li

Keyword(s):

Inflammatory Response ◽

High Glucose ◽

Ros Production ◽

Metformin Treatment ◽

Induced Apoptosis ◽

Pp2a Activation

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1,25(OH)2D3 Inhibits High Glucose-induced Apoptosis and ROS Production in Human Peritoneal Mesothelial Cells Through MAPK/P38 Pathway

Hong Kong Journal of Nephrology ◽

10.1016/j.hkjn.2015.09.204 ◽

2015 ◽

Vol 17 (2) ◽

pp. S127

Author(s):

Lina Yang ◽

Jianfei Ma

Keyword(s):

High Glucose ◽

Mesothelial Cells ◽

Ros Production ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

Induced Apoptosis ◽

P38 Pathway

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High glucose induces Nox4 expression and podocyte apoptosis through the Smad3/ezrin/PKA pathway

Biology Open ◽

10.1242/bio.055012 ◽

2020 ◽

pp. bio.055012

Author(s):

Wanxu Guo ◽

Hang Gao ◽

Wei Pan ◽

Panapn Yu ◽

Guanghua Che

Keyword(s):

High Glucose ◽

Molecular Mechanisms ◽

Ros Generation ◽

Intracellular Camp ◽

Ros Production ◽

Intracellular Ros ◽

Induced Apoptosis ◽

Pka Pathway ◽

Major Target

Podocyte is the major target in proteinuric kidney disease such as diabetic nephropathy. The underlying molecular mechanisms by which high glucose (HG) results in podocyte damage remain unclear. This study investigated the regulatory role of Smad3, ezrin, and protein kinase A (PKA) in NADPH oxidase (Nox4) expression, reactive oxidative species (ROS) production, and apoptosis in HG-treated podocytes. Human podocyte cell line was cultured and differentiated, then treated with 30 mM HG. Apoptosis and intracellular ROS level was assessed using TUNEL and DCF assay, respectively. Expressions of Nox4, phospho-Smad3Ser423/425, phospho-PKAThr197, and phospho-ezrinThr567 were evaluated using Western blotting. ELISA was used to quantify intracellular cAMP concentration and PKA activity. Knockdown assay was used to inhibit the expressions of Smad3, Nox4, and ezrin by lentiviral shRNA. In HG-treated podocytes, the level of phospho-Smad3Ser423/425 and phospho-ezrinThr567 was increased significantly, which was accompanied by the reduction of cAMP and phospho-PKAThr197. HG-induced apoptosis was significantly prevented by the Smad3 inhibitor SIS3 or shRNA-Smad3. In podocytes expressing shRNA-ezrin or shRNA-Nox4, apoptosis was remarkably mitigated following HG treatment. HG-induced upregulation of phospho-ezrinThr567 and downregulation of phospho-PKAThr197 was significantly prevented by SIS3, shRNA-ezrin or shRNA-Smad3. Forskolin, a PKA activator, significantly inhibited HG-mediated upregulation of Nox4 expression, ROS generation, and apoptosis. Additionally, an increase in the ROS level was prohibited in HG-treated podocytes with the knockdown of Nox4, Smad3, or ezrin. Taken together, our findings provided evidence that Smad3-mediated ezrin activation upregulates Nox4 expression and ROS production, by suppressing PKA activity, which may at least in part contribute to HG-induced podocyte apoptosis.

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1,25(OH)2D3 inhibits high glucose-induced apoptosis and ROS production in human peritoneal mesothelial cells via the MAPK/P38 pathway

Molecular Medicine Reports ◽

10.3892/mmr.2016.5323 ◽

2016 ◽

Vol 14 (1) ◽

pp. 839-844 ◽

Cited By ~ 14

Author(s):

LINA YANG ◽

LAN WU ◽

SHUYAN DU ◽

YE HU ◽

YI FAN ◽

...

Keyword(s):

High Glucose ◽

Mesothelial Cells ◽

Ros Production ◽

Peritoneal Mesothelial Cells ◽

Human Peritoneal Mesothelial Cells ◽

Induced Apoptosis ◽

P38 Pathway

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Effects of High Glucose Concentration on Pericyte-Like Differentiated Human Adipose-Derived Mesenchymal Stem Cells

International Journal of Molecular Sciences ◽

10.3390/ijms22094604 ◽

2021 ◽

Vol 22 (9) ◽

pp. 4604

Author(s):

Giuliana Mannino ◽

Anna Longo ◽

Florinda Gennuso ◽

Carmelina Daniela Anfuso ◽

Gabriella Lupo ◽

...

Keyword(s):

Stem Cells ◽

Mesenchymal Stem Cells ◽

Mrna Expression ◽

Inflammatory Cytokines ◽

High Glucose ◽

Angiogenic Factors ◽

Diabetic Patients ◽

Ros Production ◽

Migration Ability ◽

And Migration

A pericyte-like differentiation of human adipose-derived mesenchymal stem cells (ASCs) was tested in in vitro experiments for possible therapeutic applications in cases of diabetic retinopathy (DR) to replace irreversibly lost pericytes. For this purpose, pericyte-like ASCs were obtained after their growth in a specific pericyte medium. They were then cultured in high glucose conditions to mimic the altered microenvironment of a diabetic eye. Several parameters were monitored, especially those particularly affected by disease progression: cell proliferation, viability and migration ability; reactive oxygen species (ROS) production; inflammation-related cytokines and angiogenic factors. Overall, encouraging results were obtained. In fact, even after glucose addition, ASCs pre-cultured in the pericyte medium (pmASCs) showed high proliferation rate, viability and migration ability. A considerable increase in mRNA expression levels of the anti-inflammatory cytokines transforming growth factor-β1 (TGF-β1) and interleukin-10 (IL-10) was observed, associated with reduction in ROS production, and mRNA expression of pro-inflammatory cytokines interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), and angiogenic factors. Finally, a pmASC-induced better organization of tube-like formation by retinal endothelial cells was observed in three-dimensional co-culture. The pericyte-like ASCs obtained in these experiments represent a valuable tool for the treatment of retinal damages occurring in diabetic patients.

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CD36 promotes NLRP3 inflammasome activation via the mtROS pathway in renal tubular epithelial cells of diabetic kidneys

Cell Death and Disease ◽

10.1038/s41419-021-03813-6 ◽

2021 ◽

Vol 12 (6) ◽

Author(s):

Yanjuan Hou ◽

Qian Wang ◽

Baosheng Han ◽

Yiliang Chen ◽

Xi Qiao ◽

...

Keyword(s):

Nlrp3 Inflammasome ◽

High Glucose ◽

Inflammasome Activation ◽

Ros Production ◽

Tubular Epithelial Cells ◽

Renal Tubular Epithelial Cells ◽

Nlrp3 Inflammasome Activation ◽

Ampk Activity ◽

Renal Tubular ◽

Tubulointerstitial Inflammation

AbstractTubulointerstitial inflammation plays a key role in the pathogenesis of diabetic nephropathy (DN). Interleukin-1β (IL-1β) is the key proinflammatory cytokine associated with tubulointerstitial inflammation. The NLRP3 inflammasome regulates IL-1β activation and secretion. Reactive oxygen species (ROS) represents the main mediator of NLRP3 inflammasome activation. We previously reported that CD36, a class B scavenger receptor, mediates ROS production in DN. Here, we determined whether CD36 is involved in NLRP3 inflammasome activation and explored the underlying mechanisms. We observed that high glucose induced-NLRP3 inflammasome activation mediate IL-1β secretion, caspase-1 activation, and apoptosis in HK-2 cells. In addition, the levels of CD36, NLRP3, and IL-1β expression (protein and mRNA) were all significantly increased under high glucose conditions. CD36 knockdown resulted in decreased NLRP3 activation and IL-1β secretion. CD36 knockdown or the addition of MitoTempo significantly inhibited ROS production in HK-2 cells. CD36 overexpression enhanced NLRP3 activation, which was reduced by MitoTempo. High glucose levels induced a change in the metabolism of HK-2 cells from fatty acid oxidation (FAO) to glycolysis, which promoted mitochondrial ROS (mtROS) production after 72 h. CD36 knockdown increased the level of AMP-activated protein kinase (AMPK) activity and mitochondrial FAO, which was accompanied by the inhibition of NLRP3 and IL-1β. The in vivo experimental results indicate that an inhibition of CD36 could protect diabetic db/db mice from tubulointerstitial inflammation and tubular epithelial cell apoptosis. CD36 mediates mtROS production and NLRP3 inflammasome activation in db/db mice. CD36 inhibition upregulated the level of FAO-related enzymes and AMPK activity in db/db mice. These results suggest that NLRP3 inflammasome activation is mediated by CD36 in renal tubular epithelial cells in DN, which suppresses mitochondrial FAO and stimulates mtROS production.

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