Notoginsenoside R1 Protects Against High Glucose-Induced Cell Injury Through AMPK/Nrf2 and Downstream HO-1 Signaling

Notoginsenoside R1 (NGR1), the primary bioactive compound found in Panax notoginseng, is believed to have antihypertrophic and antiapoptotic properties, and has long been used to prevent and treat cardiovascular diseases. However, its potential role in prevention of diabetic cardiomyopathy remains unclear. The present study aimed to investigate the mechanism of NGR1 action in high glucose-induced cell injury. H9c2 cardiomyocytes were cultured in a high-glucose medium as an in-vitro model, and apoptotic cells were visualized using TUNEL staining. Expression of Nrf2 and HO-1 was measured using Western blotting or reverse transcription-quantitative PCR (RT-qPCR). The Nrf2 small interfering (si) RNA was transfected into cardiomyocytes using Opti-MEM containing Lipofectamine® RNAiMAX. NGR1 protected H9c2 cardiomyocytes from cell death, apoptosis and hypertrophy induced by high glucose concentration. Expression of auricular natriuretic peptide and brain natriuretic peptide was remarkably reduced in NGR1-treated H9C2 cells. Western blot analysis showed that high glucose concentration markedly inhibited AMPK, Nrf2 and HO-1, and this could be reversed by NGR1 treatment. However, the cardioprotective effect of NGR1 was attenuated by compound C, which reverses Nrf2 and HO-1 expression levels, suggesting that AMPK upregulates Nrf2 and HO-1 gene expression, protein synthesis and secretion. Transfection of H9C2 cells with Nrf2 siRNA markedly reduced the cardioprotective effect of NGR1 via reduced expression of HO-1. These results indicated that NGR1 attenuated high glucose-induced cell injury via AMPK/Nrf2 signaling and its downstream target, the HO-1 pathway. We conclude that the cardioprotective effects of NGR1 result from upregulation of AMPK/Nrf2 signaling and HO-1 expression in cardiomyocytes. Our findings suggest that NGR1 treatment might provide a novel therapy for diabetic cardiomyopathy.

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Resveratrol inhibits high-glucose-induced inflammatory “metabolic memory” in human retinal vascular endothelial cells through SIRT1-dependent signaling

Canadian Journal of Physiology and Pharmacology ◽

10.1139/cjpp-2019-0201 ◽

2019 ◽

Vol 97 (12) ◽

pp. 1141-1151 ◽

Cited By ~ 4

Author(s):

Tingting Jiang ◽

Junxiang Gu ◽

Wenwen Chen ◽

Qing Chang

Keyword(s):

Endothelial Cells ◽

Inflammatory Cytokines ◽

High Glucose ◽

Glucose Concentration ◽

Vascular Endothelial Cells ◽

Sirtuin 1 ◽

Metabolic Memory ◽

Vascular Endothelial ◽

High Glucose Concentration ◽

Compound C

Diabetes induces vascular endothelial damage and this study investigated high-glucose-induced inflammation “metabolic memory” of human retinal vascular endothelial cells (HRVECs), the effects of resveratrol on HRVECs, and the underlying signaling. HRVECs were grown under various conditions and assayed for levels of sirtuin 1 (SIRT1); acetylated nuclear factor κB (Ac-NF-κB); NOD-like receptor family, pyrin domain containing 3 (NLRP3); and other inflammatory cytokines; and cell viability. A high glucose concentration induced HRVEC inflammation metabolic memory by decreasing SIRT1 and increasing Ac-NF-κB, NLRP3, caspase 1, interleukin-1β, inducible nitric oxide synthase, and tumor necrosis factor α, whereas exposure of HRVECs to a high glucose medium for 4 days, followed by a normal glucose concentration for an additional 4 days, failed to reverse these changes. A high glucose concentration also significantly reduced HRVEC viability. In contrast, resveratrol, a selective SIRT1 activator, markedly enhanced HRVEC viability and reduced the inflammatory cytokines expressions. In addition, high glucose reduced AMP-activated protein kinase (AMPK) phosphorylation and retained during the 4 days of the reversal period of culture. The effects of resveratrol were abrogated after co-treatment with the SIRT1 inhibitor nicotinamide and the AMPK inhibitor compound C. In conclusion, resveratrol was able to reverse high-glucose-induced inflammation “metabolic memory” of HRVECs by activation of the SIRT1/AMPK/NF-κB pathway.

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Lipopolysaccharide (LPS) Aggravates High Glucose- and Hypoxia/Reoxygenation-Induced Injury through Activating ROS-Dependent NLRP3 Inflammasome-Mediated Pyroptosis in H9C2 Cardiomyocytes

Journal of Diabetes Research ◽

10.1155/2019/8151836 ◽

2019 ◽

Vol 2019 ◽

pp. 1-12 ◽

Cited By ~ 14

Author(s):

Zhen Qiu ◽

Yuhong He ◽

Hao Ming ◽

Shaoqing Lei ◽

Yan Leng ◽

...

Keyword(s):

Cell Viability ◽

Nlrp3 Inflammasome ◽

High Glucose ◽

Cell Injury ◽

H9c2 Cell ◽

Ros Production ◽

H9c2 Cells ◽

Caspase 1 ◽

H9c2 Cardiomyocytes ◽

Hypoxia Reoxygenation

Diabetes aggravates myocardial ischemia-reperfusion (I/R) injury because of the combination effects of changes in glucose and lipid energy metabolism, oxidative stress, and systemic inflammatory response. Studies have indicated that myocardial I/R may coincide and interact with sepsis and inflammation. However, the role of LPS in hypoxia/reoxygenation (H/R) injury in cardiomyocytes under high glucose conditions is still unclear. Our objective was to examine whether lipopolysaccharide (LPS) could aggravate high glucose- (HG-) and hypoxia/reoxygenation- (H/R-) induced injury by upregulating ROS production to activate NLRP3 inflammasome-mediated pyroptosis in H9C2 cardiomyocytes. H9C2 cardiomyocytes were exposed to HG (30 mM) condition with or without LPS, along with caspase-1 inhibitor (Ac-YVAD-CMK), inflammasome inhibitor (BAY11-7082), ROS scavenger N-acetylcysteine (NAC), or not for 24 h, then subjected to 4 h of hypoxia followed by 2 h of reoxygenation (H/R). The cell viability, lactate dehydrogenase (LDH) release, caspase-1 activity, and intracellular ROS production were detected by using assay kits. The incidence of pyroptosis was detected by calcein-AM/propidium iodide (PI) double staining kit. The concentrations of IL-1β and IL-18 in the supernatants were assessed by ELISA. The mRNA levels of NLRP3, ASC, and caspase-1 were detected by qRT-PCR. The protein levels of NF-κB p65, NLRP3, ASC, cleaved caspase-1 (p10), IL-1β, and IL-18 were detected by western blot. The results indicated that pretreatment LPS with 1 μg/ml not 0.1 μg/ml could efficiently aggravate HG and H/R injury by activating NLRP3 inflammasome to mediate pyroptosis in H9C2 cells, as evidenced by increased LDH release and decreased cell viability in the cells, and increased expression of NLRP3, ASC, cleaved caspase-1 (p10), IL-1β, and IL-18. Meanwhile, Ac-YVAD-CMK, BAY11-7082, or NAC attenuated HG- and H/R-induced H9C2 cell injury with LPS stimulated by reversing the activation of NLRP3 inflammasome-mediated pyroptosis. In conclusion, LPS could increase the sensitivity of H9C2 cells to HG and H/R and aggravated HG- and H/R-induced H9C2 cell injury by promoting ROS production to induce NLRP3 inflammasome-mediated pyroptosis.

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Stimulated disruption of Cav‐1/eNOS association plays a vital role in adaptation of H9c2 cells against high glucose concentration via Akt /HSP90 /eNOS activation.

The FASEB Journal ◽

10.1096/fasebj.22.1_supplement.1226.10 ◽

2008 ◽

Vol 22 (S1) ◽

Author(s):

Samson Mathews Samuel ◽

Suresh Varma Penumathsa ◽

Perumana R Sudhakaran ◽

Nilanjana Maulik

Keyword(s):

High Glucose ◽

Glucose Concentration ◽

Vital Role ◽

High Glucose Concentration ◽

H9c2 Cells

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Long noncoding RNA OIP5-AS1 overexpression promotes viability and inhibits high glucose-induced oxidative stress of cardiomyocytes by targeting microRNA-34a/SIRT1 axis in diabetic cardiomyopathy

Endocrine Metabolic & Immune Disorders - Drug Targets ◽

10.2174/1871530321666201230090742 ◽

2020 ◽

Vol 21 ◽

Author(s):

Haiyun Sun ◽

Chong Wang ◽

Ying Zhou ◽

Xingbo Cheng

Keyword(s):

Oxidative Stress ◽

Protein Expression ◽

Diabetic Cardiomyopathy ◽

High Glucose ◽

Long Noncoding Rna ◽

Noncoding Rna ◽

Luciferase Reporter ◽

H9c2 Cells ◽

Promoting Effect

Objective: Diabetic cardiomyopathy (DCM) is an important complication of diabetes. This study was attempted to discover the effects of long noncoding RNA OIP5-AS1 (OIP5-AS1) on the viability and oxidative stress of cardiomyocyte in DCM. Methods: The expression of OIP5-AS1 and microRNA-34a (miR-34a) in DCM was detected by qRT-PCR. In vitro, DCM was simulated by high glucose (HG, 30 mM) treatment in H9c2 cells. The viability of HG (30 mM)-treated H9c2 cells was examined by MTT assay. The reactive oxygen species (ROS), superoxide dismutase (SOD) and malondialdehyde (MDA) levels were used to evaluate the oxidative stress of HG (30 mM)-treated H9c2 cells. Dual-luciferase reporter assay was used to confirm the interactions among OIP5-AS1, miR-34a and SIRT1. Western blot was applied to analyze the protein expression of SIRT1. Results: The expression of OIP5-AS1 was down-regulated in DCM, but miR-34a was up-regulated. The functional experiment stated that OIP5-AS1 overexpression increased the viability and SOD level, while decreased the ROS and MDA levels in HG (30 mM)-treated H9c2 cells. The mechanical experiment confirmed that OIP5-AS1 and SIRT1 were both targeted by miR-34a with the complementary binding sites at 3′UTR. MiR-34a overexpression inhibited the protein expression of SIRT1. In the feedback experiments, miR-34a overexpression or SIRT1 inhibition weakened the promoting effect on viability, and mitigated the reduction effect on oxidative stress caused by OIP5-AS1 overexpression in HG (30 mM)-treated H9c2 cells. Conclusions: OIP5-AS1 overexpression enhanced viability and attenuated oxidative stress of cardiomyocyte via regulating miR-34a/SIRT1 axis in DCM, providing a new therapeutic target for DCM.

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High glucose concentration augments angiotensin II mediated contraction via AT1 receptors in rat thoracic aorta

Pharmacological Research ◽

10.1016/j.phrs.2004.06.001 ◽

2004 ◽

Vol 50 (6) ◽

pp. 561-568 ◽

Cited By ~ 18

Author(s):

K ARUN ◽

C KAUL ◽

P RAMARAO

Keyword(s):

Angiotensin Ii ◽

Thoracic Aorta ◽

High Glucose ◽

Glucose Concentration ◽

High Glucose Concentration ◽

At1 Receptors ◽

Rat Thoracic Aorta

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High glucose concentration suppresses mesangial laminin B2 gene expression

Journal of Diabetic Complications ◽

10.1016/0891-6632(91)90039-r ◽

1991 ◽

Vol 5 (2-3) ◽

pp. 118-120 ◽

Cited By ~ 2

Author(s):

Shigehiro Katayama ◽

Mari Abe ◽

Kiyoshi Tanaka ◽

Akira Omoto ◽

Kiyohiko Negishi ◽

...

Keyword(s):

Gene Expression ◽

High Glucose ◽

Glucose Concentration ◽

High Glucose Concentration

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Sulodexide reduces glucose induced senescence in human retinal endothelial cells

Scientific Reports ◽

10.1038/s41598-021-90987-w ◽

2021 ◽

Vol 11 (1) ◽

Author(s):

A. Gericke ◽

K. Suminska-Jasińska ◽

A. Bręborowicz

Keyword(s):

Endothelial Cells ◽

High Glucose ◽

Glucose Concentration ◽

Chronic Exposure ◽

Replicative Senescence ◽

Population Doubling ◽

Population Doubling Time ◽

High Glucose Concentration ◽

Retinal Endothelial Cells

AbstractChronic exposure of retinal endothelium cells to hyperglycemia is the leading cause of diabetic retinopathy. We evaluated the effect of high glucose concentration on senescence in human retinal endothelial cells (HREC) and modulation of that effect by Sulodexide. Experiments were performed on HREC undergoing in vitro replicative senescence in standard medium or medium supplemented with glucose 20 mmol/L (GLU) or mannitol 20 mnol/L (MAN). Effect of Sulodexide 0.5 LRU/mL (SUL) on the process of HREC senescence was studied. Glucose 20 mmol/L accelerates senescence of HREC: population doubling time (+ 58%, p < 0.001) β-galactosidase activity (+ 60%, p < 0.002) intracellular oxidative stress (+ 65%, p < 0.01), expression of p53 gene (+ 118%, p < 0.001). Senescent HREC had also reduced transendothelial electrical resistance (TEER) (− 30%, p < 0.001). Mannitol 20 mmol/L used in the same scenario as glucose did not induce HREC senescence. In HREC exposed to GLU and SUL, the senescent changes were smaller. HREC, which became senescent in the presence of GLU, demonstrated higher expression of genes regulating the synthesis of Il6 and VEGF-A, which was reflected by increased secretion of these cytokines (IL6 + 125%, p < 0.001 vs control and VEGF-A + 124% p < 0.001 vs control). These effects were smaller in the presence of SUL, and additionally, an increase of TEER in the senescent HREC was observed. Chronic exposure of HREC to high glucose concentration in medium accelerates their senescence, and that process is reduced when the cells are simultaneously exposed to Sulodexide. Additionally, Sulodexide decreases the secretion of IL6 and VEGF-A from senescent HREC and increases their TEER.

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The influence of high glucose concentration on the ability of mesenchymal stromal cells to stimulate blood vessel growth

Diabetes Mellitus ◽

10.14341/2072-0351-5631 ◽

2011 ◽

Vol 14 (2) ◽

pp. 32-35 ◽

Cited By ~ 2

Author(s):

Zhanna Alekseevna Akopyan ◽

Georgy Vladimirovich Sharonov ◽

Tatiana Nikolaevna Kochegura ◽

Natalya Fedorovna Il'yashenko ◽

Igor Eduardovich Belyanko ◽

...

Keyword(s):

Adipose Tissue ◽

Blood Vessel ◽

High Glucose ◽

Glucose Concentration ◽

Functional Activity ◽

Human Adipose Tissue ◽

Diabetic Patients ◽

High Glucose Concentration ◽

Vessel Growth ◽

Negative Effect

Adipose issue is a source of mesenchymal stem cells (MSC) that can be used to stimulate blood vessel growth in ischemic tissues. Various metabolicdisorders including hypeglycemia may have negative effect on therapeutic properties of these cells. Aim. To study the influence of high glucose concentration on functional activity in human adipose tissue. Materials and methods. Flow cytometry and real time PCR were used to study functional activity of cultured MSC from human adipose issue at highglucose concentration. Results. Prolonged (10-12 days) incubation at a high glucose concentration (25 mM) suppressed the ability of MSC to stimulate angiogenesis. Also,glucose modified expression of genes activating and inhibiting angiogenesis but had no effect on MSC proliferation and apoptosis. Conclusion. High glucose concentration suppresses angiogenic activity of MSC in adipose tissue; it may account for incomplete restoration of bloodflow in diabetic patients.

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