Carnosine Protects Mouse Podocytes from High Glucose Induced Apoptosis through PI3K/AKT and Nrf2 Pathways

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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Circ-ACTR2 aggravates the high glucose-induced cell dysfunction of human renal mesangial cells through mediating the miR-205-5p/HMGA2 axis in diabetic nephropathy

Diabetology & Metabolic Syndrome ◽

10.1186/s13098-021-00692-x ◽

2021 ◽

Vol 13 (1) ◽

Author(s):

Jie Yun ◽

Jinyu Ren ◽

Yufei Liu ◽

Lijuan Dai ◽

Liqun Song ◽

...

Keyword(s):

Oxidative Stress ◽

Diabetic Nephropathy ◽

High Glucose ◽

Cell Injury ◽

Mesangial Cells ◽

Protein Detection ◽

Cell Counting ◽

Luciferase Reporter ◽

Enzyme Linked Immunosorbent Assay ◽

Cell Dysfunction

Abstract Background Circular RNAs (circRNAs) have been considered as pivotal biomarkers in Diabetic nephropathy (DN). CircRNA ARP2 actin-related protein 2 homolog (circ-ACTR2) could promote the HG-induced cell injury in DN. However, how circ-ACTR2 acts in DN is still unclear. This study aimed to explore the molecular mechanism of circ-ACTR2 in DN progression, intending to provide support for the diagnostic and therapeutic potentials of circ-ACTR2 in DN. Methods RNA expression analysis was conducted by the quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Cell growth was measured via Cell Counting Kit-8 and EdU assays. Inflammatory response was assessed by Enzyme-linked immunosorbent assay. The protein detection was performed via western blot. Oxidative stress was evaluated by the commercial kits. The molecular interaction was affirmed through dual-luciferase reporter and RNA immunoprecipitation assays. Results Circ-ACTR2 level was upregulated in DN samples and high glucose (HG)-treated human renal mesangial cells (HRMCs). Silencing the circ-ACTR2 expression partly abolished the HG-induced cell proliferation, inflammation and extracellular matrix accumulation and oxidative stress in HRMCs. Circ-ACTR2 was confirmed as a sponge for miR-205-5p. Circ-ACTR2 regulated the effects of HG on HRMCs by targeting miR-205-5p. MiR-205-5p directly targeted high-mobility group AT-hook 2 (HMGA2), and HMGA2 downregulation also protected against cell injury in HG-treated HRMCs. HG-mediated cell dysfunction was repressed by miR-205-5p/HMGA2 axis. Moreover, circ-ACTR2 increased the expression of HMGA2 through the sponge effect on miR-205-5p in HG-treated HRMCs. Conclusion All data have manifested that circ-ACTR2 contributed to the HG-induced DN progression in HRMCs by the mediation of miR-205-5p/HMGA2 axis.

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Norcantharidin-induced Apoptosis of AGS Human Gastric Cancer Cells Through Reactive Oxygen Species Production, and Caspase- and Mitochondria-dependent Signaling Pathways

Anticancer Research ◽

10.21873/anticanres.11192 ◽

2016 ◽

Vol 36 (11) ◽

pp. 6031-6042 ◽

Cited By ~ 10

Author(s):

LI-CHENG ZHENG ◽

MEI-DUE YANG ◽

CHAO-LIN KUO ◽

CHIA-HSIN LIN ◽

MING-JEN FAN ◽

...

Keyword(s):

Gastric Cancer ◽

Reactive Oxygen Species ◽

Cancer Cells ◽

Signaling Pathways ◽

Reactive Oxygen Species Production ◽

Human Gastric Cancer ◽

Oxygen Species ◽

Gastric Cancer Cells ◽

Induced Apoptosis ◽

Species Production

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ALK7 Inhibition Protects Osteoblast Cells Against High Glucose-induced ROS Production via Nrf2/HO-1 Signaling Pathway

Current Molecular Medicine ◽

10.2174/1566524021666210614144337 ◽

2021 ◽

Vol 21 ◽

Author(s):

Zhen Zhao ◽

Yu Lu ◽

Huan Wang ◽

Xiang Gu ◽

Luting Zhu ◽

...

Keyword(s):

Oxidative Stress ◽

Cell Viability ◽

Signaling Pathway ◽

High Glucose ◽

Collagen I ◽

Cell Counting ◽

Enzyme Linked Immunosorbent Assay ◽

Heme Oxygenase 1 ◽

Ros Production ◽

Alizarin Red

Background: Some studies demonstrated that under high-glucose (HG) condition, osteoblasts develop oxidative stress, which will impair their normal functions. The effects of activin receptor-like kinase 7 (ALK7) silencing on HG-induced osteoblasts remained unclear. Objective: The aim of this study was to explore the effect of ALK7 on HG-induced osteoblasts. Methods: MC3T3-E1 cells were treated with different concentrations of HG (0, 50, 100, 200 and 300mg/dL), and the cell viability was detected using cell counting kit-8 (CCK-8). HG-treated MC3T3-E1 cells were transfected with siALK7 or ALK7 overexpression plasmid or siNrf2, and then the viability and apoptosis were detected by CCK-8 and flow cytometry. The levels of reactive oxygen species (ROS), collagen I and calcification nodule were determined by oxidative stress kits, Enzyme-linked immunosorbent assay and Alizarin red staining. The expressions of NF-E2-related factor 2 (Nrf2), heme oxygenase-1 (HO-1) and osteoblast-associated genes were determined by quantitative real-time PCR (qRT-PCR) and Western blot. Results: Cell viability was reduced with HG treatment. Silencing ALK7 inhibited the effect of HG on increasing cell apoptosis and ROS production, reduced cell viability, mineralized nodules, and downregulated collagen I and osteoblast-associated genes expression in MC3T3-E1 cells. ALK7 silencing activated the Nrf2/HO-1 signaling pathway by affecting expressions of HO-1 and Nrf2. ALK7 overexpression had the opposite effects. In addition, siNrf2 partially reversed the effects of ALK7 silencing on HG-induced MC3T3-E1 cells. Conclusion: ALK7 silencing protected osteoblasts under HG condition possibly through activating the Nrf2/HO-1 pathway.

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Abstract 098: MicroRNA Regulation of NADPH Oxidase Mediates the Antioxidant Effect of Renal Paraoxonase 2

Hypertension ◽

10.1161/hyp.66.suppl_1.098 ◽

2015 ◽

Vol 66 (suppl_1) ◽

Author(s):

Santiago Cuevas ◽

Yu Yang ◽

Laureano D Asico ◽

John Jones ◽

Ines Armando ◽

...

Keyword(s):

Nadph Oxidase ◽

Antioxidant Effect ◽

Receptor Subtype ◽

Ros Production ◽

Oxygen Species ◽

Wild Type ◽

Compensatory Response ◽

Microrna Regulation ◽

Dopamine 2 Receptor ◽

Renal Expression

Increased renal generation of reactive oxygen species (ROS) is important in the pathogenesis of hypertension caused by absent or dysfunctional dopamine receptor subtype. Germline deletion of the dopamine 2 receptor in mice increases renal NADPH oxidase (NOX) activity and decreases expression paraoxonase 2 (PON2) and results in ROS-dependent hypertension. We determined if microRNA (miR) is involved in PON2-mediated regulation of NOX. Silencing PON2 in human renal proximal tubules cells decreased PON2 (-60±4%, n=3,*P<0.05) and increased NOX2 (110±15%, n=3,*P<0.05) and NOX4 (80±10%, n=3,*P<0.05) proteins, NOX activity (50±6%, n=3,*P<0.05), and ROS production (57±3%, n=4,*P<0.05). Inhibition of NOX activity by diphenyleneiodonium normalized the increase in ROS caused by PON2 silencing. Renal-selective silencing of Pon2 in mice by the renal subcapsular infusion of Pon2 siRNA decreased PON2 (~50%), and increased NOX2 (191±11%,n=3, P<0.05), NOX4 (60±4%,n=3, P<0.05), NOX activity (94±23%, n=3, P<0.05), and blood pressure (BP) (+41±6 mmHg, n=3, P<0.05). Pon2-/- mice also had higher BP than wild-type littermates (+15±2 mmHg,n=3/4,*P<0.05) but less than observed with renal-selective silencing indicating extrarenal compensation. Renal NOX2 (220±64%, n=3/4,*P<0.05) and NOX activity (195±77%, n=3,*P<0.05) were also increased in Pon2-/- mice. However, the renal expression of NOX4 was similar in Pon2-/- and wild-type littermates. The renal expressions of miR-23b, miR-34a and miR-155 (reported to regulate NOX expression) were also similar in Pon2-/- mice and wild-type littermates. However, renal miR-146a expression was decreased (-25±4%, n=3/4,*P<0.05) while miR-204 (150±12%, n=3/4,*P<0.05) and NFAT expressions (21±7%, n=3/4,*P<0.05) were increased in Pon2-/- mice. The increase in miR-204 could be a compensatory response because miR-204 has been shown to decrease NFAT expression. It is known that NFAT and NOX2 can positively regulate each other’s expression while miR-146a negatively regulates NOX4 expression and inflammation. We conclude that PON2 by increasing miR-146a and decreasing NFAT expression negatively regulates NOX activity and reduce ROS production that would contribute to the maintenance of normal BP.

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Clinical Isolates ofMycobacterium tuberculosisDiffer in Their Ability to Induce Respiratory Burst and Apoptosis in Neutrophils as a Possible Mechanism of Immune Escape

Clinical and Developmental Immunology ◽

10.1155/2012/152546 ◽

2012 ◽

Vol 2012 ◽

pp. 1-11 ◽

Cited By ~ 18

Author(s):

María M. Romero ◽

Luciana Balboa ◽

Juan I. Basile ◽

Beatriz López ◽

Viviana Ritacco ◽

...

Keyword(s):

Reactive Oxygen Species ◽

Reference Strain ◽

Immune Escape ◽

Multidrug Resistant ◽

Polymorphonuclear Neutrophils ◽

Ros Production ◽

Oxygen Species ◽

Phenotypic Differences ◽

Induced Apoptosis ◽

P38 Pathway

Tuberculosis pathogenesis was earlier thought to be mainly related to the host but now it appears to be clear that bacterial factors are also involved. Genetic variability ofMycobacterium tuberculosis(Mtb) could be slight but it may lead to sharp phenotypic differences. We have previously reported that nonopsonizedMtbH37Rv induce apoptosis of polymorphonuclear neutrophils (PMNs) by a mechanism that involves the p38 pathway. Here we evaluated the capability to induce PMN apoptosis of two prevalentMtblineages in Argentina, the Latin America and Mediterranean (LAM), and Haarlem, using the H37Rv as a reference strain. Results showed that LAM strains strongly induced apoptosis of PMN which correlated with the induction of reactive oxygen species (ROS) production and p38 activation. Interestingly, the highly prosperous multidrug-resistant M strain, belonging to the Haarlem lineage, lacked the ability to activate and to induce PMN apoptosis as a consequence of (1) a weak ROS production and (2) the contribution of antiapoptotic mechanisms mediated at least by ERK. Although with less skill, M is able to enter the PMN so that phenotypic differences could lead PMN to be a reservoir allowing some pathogens to prevail and persist over other strains in the community.

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