scholarly journals Cissus quadrangularis Linn. Extract Reduces High Glucose Induced Inhibition of In Vitro Proliferation and Matrix Mineralization in MG-63 Cells

2018 ◽  
Vol 16 (2) ◽  
pp. 159-164
Author(s):  
Madhu Gupta ◽  
Md Arshad
Keyword(s):  
Cell Viability ◽  
High Glucose ◽  
Reactive Oxygen Species Generation ◽  
Ethanolic Extract ◽  
Nodule Formation ◽  
Physiological Control ◽  
In Vitro Proliferation ◽  
Matrix Mineralization ◽  
Cissus Quadrangularis

The aim of this study was to evaluate the effect and mechanism of action of ethanolic extract of Cissus quadrangularis Linn. stem in an in vitro system using MG-63 (osteoblast-like) cells. Cells were pre-exposed to high glucose (45 mmol/l). Insulin was used as positive control. To determine whether C. quadrangularis (CQ) extract could prevent hyperglycaemia induced oxidative stress, and apoptosis we performed reactive oxygen species generation using 2’,7’-dichlorofluorescin diacetate and propidium iodide staining for apoptosis. For cell proliferation, we carried out cell viability (MTT) assay. Further, alkaline phosphate assay and alizarin staining were done to study the matrix mineralization. High glucose (45 mmol/l) significantly (P < 0.001) inhibited the cell viability and matrix mineralization, and induced apoptosis as compared to physiological control. C. quadrangularis extract as well as insulin treatment significantly (p < 0.001 versus high glucose) restored the cell viability, cell differentiation and bone nodule formation. Thus, ethanolic extract of C. quadrangularis stem prevents the antiproliferative effect of high glucose.Dhaka Univ. J. Pharm. Sci. 16(2): 159-164, 2017 (December)

scholarly journals Metformin Protects H9C2 Cardiomyocytes from High-Glucose and Hypoxia/Reoxygenation Injury via Inhibition of Reactive Oxygen Species Generation and Inflammatory Responses: Role of AMPK and JNK

2016 ◽  
Vol 2016 ◽  
pp. 1-9 ◽  
Author(s):  
Mingyan Hu ◽  
Ping Ye ◽  
Hua Liao ◽  
Manhua Chen ◽  
Feiyan Yang
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Viability ◽  
High Glucose ◽  
Molecular Mechanisms ◽  
Ischemia Reperfusion ◽  
Reactive Oxygen Species Generation ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Compound C ◽  
Hypoxia Reoxygenation

Metformin is a first-line drug for the management of type 2 diabetes. Recent studies suggested cardioprotective effects of metformin against ischemia/reperfusion injury. However, it remains elusive whether metformin provides direct protection against hypoxia/reoxygenation (H/R) injury in cardiomyocytes under normal or hyperglycemic conditions. This study in H9C2 rat cardiomyoblasts was designed to determine cell viability under H/R and high-glucose (HG, 33 mM) conditions and the effects of cotreatment with various concentrations of metformin (0, 1, 5, and 10 mM). We further elucidated molecular mechanisms underlying metformin-induced cytoprotection, especially the possible involvement of AMP-activated protein kinase (AMPK) and Jun NH(2)-terminal kinase (JNK). Results indicated that 5 mM metformin improved cell viability, mitochondrial integrity, and respiratory chain activity under HG and/or H/R (P<0.05). The beneficial effects were associated with reduced levels of reactive oxygen species generation and proinflammatory cytokines (TNF-α, IL-1α, and IL-6) (P<0.05). Metformin enhanced phosphorylation level of AMPK and suppressed HG + H/R induced JNK activation. Inhibitor of AMPK (compound C) or activator of JNK (anisomycin) abolished the cytoprotective effects of metformin. In conclusion, our study demonstrated for the first time that metformin possessed direct cytoprotective effects against HG and H/R injury in cardiac cells via signaling mechanisms involving activation of AMPK and concomitant inhibition of JNK.

MicroRNA-204-3p Attenuates High Glucose-Induced MPC5 Podocytes Apoptosis by Targeting Braykinin B2 Receptor

2018 ◽  
Vol 127 (06) ◽  
pp. 387-395 ◽  
Author(s):  
Xu Han ◽  
Qiaobei Li ◽  
Chunyan Wang ◽  
Yinyan Li
Keyword(s):  
Cell Viability ◽  
High Glucose ◽  
Protective Role ◽  
Luciferase Reporter ◽  
Gene And Protein Expression ◽  
Glucose Treatment ◽  
Apoptosis Gene ◽  
Induced Apoptosis

Abstract Background Previous study has been reported that braykinin B2 receptor (Bdkrb2) involves in high glucose-induced renal and podocytes injuries. However, there have been some studies with contradictory results that Bdkrb2 has a protective effect on hyperglycemia-induced injuries in vivo and in vitro. The purpose of the present study was carried out to further investigate the post-transcriptional regulatory mechanism of microRNA (miR) in high glucose-treated podocytes by targeting Bdkrb2 signaling in vitro. Methods The CCK-8 and flow cytometry were performed to measure the cell viability and apoptosis. Gene and protein expression were assayed by RT-qPCR and western blotting, respectively. Results High glucose treatment decreased cell viability and induced membrane and DNA damage, as well as apoptosis in podocytes. High glucose treatment also increased the expression of Bdkrb2, which was blocked by miR-204-3p mimics transfection in podocytes. Bioinformatics and luciferase reporter activity showed that miR-204-3p was directly targeted to the 3′-untranslated region (3′-UTR) of Bdkrb2. High glucose-induced apoptosis and dysfunction in podocytes were reserved by miR-204-3p mimics transfection, while the effects of miR-204-3p mimics in high glucose-treated podocytes were neutralized by overexpressed Bdkrb2. Conclusions These findings suggested that miR-204-3p may play a protective role in high glucose-induced apoptosis and dysfunction in podocytes through down-regulation of Bdkrb2.

scholarly journals PHNQ from Evechinus chloroticus Sea Urchin Supplemented with Calcium Promotes Mineralization in Saos-2 Human Bone Cell Line

Marine Drugs ◽  
2020 ◽  
Vol 18 (7) ◽  
pp. 373
Author(s):  
Yakun Hou ◽  
Alan Carne ◽  
Michelle McConnell ◽  
Sonya Mros ◽  
Elena A. Vasileva ◽  
...  
Keyword(s):  
Cell Viability ◽  
Sea Urchin ◽  
Xylenol Orange ◽  
Bone Cell ◽  
Future Research ◽  
Nodule Formation ◽  
Dependent Manner ◽  
Cell Viability Assay ◽  
Echinochrome A

Polyhydroxylated naphthoquinones (PHNQs), known as spinochromes that can be extracted from sea urchins, are bioactive compounds reported to have medicinal properties and antioxidant activity. The MTT (3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) cell viability assay showed that pure echinochrome A exhibited a cytotoxic effect on Saos-2 cells in a dose-dependent manner within the test concentration range (15.625–65.5 µg/mL). The PHNQ extract from New Zealand sea urchin Evechinus chloroticus did not induce any cytotoxicity within the same concentration range after 21 days of incubation. Adding calcium chloride (CaCl2) with echinochrome A increased the number of viable cells, but when CaCl2 was added with the PHNQs, cell viability decreased. The effect of PHNQs extracted on mineralized nodule formation in Saos-2 cells was investigated using xylenol orange and von Kossa staining methods. Echinochrome A decreased the mineralized nodule formation significantly (p < 0.05), while nodule formation was not affected in the PHNQ treatment group. A significant (p < 0.05) increase in mineralization was observed in the presence of PHNQs (62.5 µg/mL) supplemented with 1.5 mM CaCl2. In conclusion, the results indicate that PHNQs have the potential to improve the formation of bone mineral phase in vitro, and future research in an animal model is warranted.

In- vitro Antioxidant and Cardio-protective effect of Delonix elata (L.) Leaf extract against Doxorubicin induced toxicity in H9c2 Cardio-myocyte cell line

2021 ◽  
pp. 5635-5641
Author(s):  
Archana V ◽  
Indumathy R
Keyword(s):  
Reactive Oxygen Species ◽  
Protective Effect ◽  
Cell Viability ◽  
Leaf Extract ◽  
Radical Scavenging ◽  
Reactive Oxygen ◽  
Ethanolic Extract ◽  
Antioxidant Property ◽  
Oxygen Species

Objective: The aim of this study is to evaluate the protective effect of Delonix elata (L.) leaf extract against doxorubicin-induced cardiotoxicity in H9c2 cells. Methods: Doxorubicin has been used to treat cancer, but its clinical uses are limited because of its dose-dependent cardiotoxicity. Reactive oxygen species play an important role in the pathological process of cardiotoxicity. The various extracts (pet.ether, ethyl acetate and ethanol) of Delonix elata leaves antioxidant property was evaluated by SOD antioxidant assay and DPPH free radical scavenging assay. The cells were incubated with different concentrations of various extracts of Delonix elata leaves for 2 hr, followed by incubation with 5µM doxorubicin for 24 hr. Cell viability was determined by using MTT assay, respectively. Results: The various extracts of Delonix elata leaves exhibits antioxidant activity. The Doxorubicin significantly decreased cell viability which was accompanied by an increased ROS production. Pre-treatment with various extracts of Delonix elata leaves increased the viability ofcells and inhibit the generation of reactive oxygen species. Conclusion: In this study, findings how that Delonix elata leaf extract exhibited a protective effect against oxidative stress-induced cardiomyocyte damage. The ethanolic extract of Delonix elata leaves possesses significant antioxidant and cardioprotective activity.

scholarly journals Renoprotective Effects of Aldose Reductase Inhibitor Epalrestat against High Glucose-Induced Cellular Injury

2017 ◽  
Vol 2017 ◽  
pp. 1-11 ◽  
Author(s):  
Heba El Gamal ◽  
Ali Hussein Eid ◽  
Shankar Munusamy
Keyword(s):  
Cell Cycle ◽  
Cell Viability ◽  
Aldose Reductase ◽  
High Glucose ◽  
Mitochondrial Membrane ◽  
Cell Injury ◽  
Renal Tubular ◽  
Stage Renal Disease ◽  
End Stage

Diabetic nephropathy (DN) is the leading cause of end stage renal disease worldwide. Increased glucose flux into the aldose reductase (AR) pathway during diabetes was reported to exert deleterious effects on the kidney. The objective of this study was to investigate the renoprotective effects of AR inhibition in high glucose milieu in vitro. Rat renal tubular (NRK-52E) cells were exposed to high glucose (30 mM) or normal glucose (5 mM) media for 24 to 48 hours with or without the AR inhibitor epalrestat (1 μM) and assessed for changes in Akt and ERK1/2 signaling, AR expression (using western blotting), and alterations in mitochondrial membrane potential (using JC-1 staining), cell viability (using MTT assay), and cell cycle. Exposure of NRK-52E cells to high glucose media caused acute activation of Akt and ERK pathways and depolarization of mitochondrial membrane at 24 hours. Prolonged high glucose exposure (for 48 hours) induced AR expression andG1 cell cycle arrest and decreased cell viability (84% compared to control) in NRK-52E cells. Coincubation of cells with epalrestat prevented the signaling changes and renal cell injury induced by high glucose. Thus, AR inhibition represents a potential therapeutic strategy to prevent DN.

scholarly journals Propolis, a Constituent of Honey, Inhibits the Development of Sugar Cataracts and High-Glucose-Induced Reactive Oxygen Species in Rat Lenses

2016 ◽  
Vol 2016 ◽  
pp. 1-6 ◽  
Author(s):  
Teppei Shibata ◽  
Shinsuke Shibata ◽  
Naoko Shibata ◽  
Etsuko Kiyokawa ◽  
Hiroshi Sasaki ◽  
...  
Keyword(s):  
Reactive Oxygen Species ◽  
Cell Death ◽  
Cell Viability ◽  
High Glucose ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Histochemical Analysis ◽  
Intracellular Ros ◽  
Sugar Cataract

Purpose.This study investigated the effects of oral propolis on the progression of galactose-induced sugar cataracts in rats and thein vitroeffects of propolis on high-glucose-induced reactive oxygen species (ROS) and cell death in cultured rat lens cells (RLECs).Methods. Galactose-fed rats and RLECs cultured in high glucose (55 mM) medium were treated with propolis or vehicle control. Relative lens opacity was assessed by densitometry and changes in lens morphology by histochemical analysis. Intracellular ROS levels and cell viability were measured.Results. Oral administration of propolis significantly inhibited the onset and progression of cataract in 15% and 25% of galactose-fed rats, respectively. RLECs cultured with high glucose showed a significant increase in ROS expression with reduced cell viability. Treatment of these RLECs with 5 and 50 μg/mL propolis cultured significantly reduced ROS levels and increased cell viability, indicating that the antioxidant activity of propolis protected cells against ROS-induced damage.Conclusion. Propolis significantly inhibited the onset and progression of sugar cataract in rats and mitigated high-glucose-induced ROS production and cell death. These effects may be associated with the ability of propolis to inhibit hyperglycemia-evoked oxidative or osmotic stress-induced cellular insults.

scholarly journals Bioactive Glass Particles in Two-Dimensional and Three-Dimensional Osteogenic Cell Cultures

2017 ◽  
Vol 28 (3) ◽  
pp. 307-316 ◽  
Author(s):  
Luciana B. Alves ◽  
Sérgio L. S. de Souza ◽  
Mario Taba Jr ◽  
Arthur B. Novaes Jr ◽  
Paulo T. de Oliveira ◽  
...  
Keyword(s):  
Cell Viability ◽  
Bioactive Glass ◽  
Bone Matrix ◽  
Three Dimensional ◽  
Nodule Formation ◽  
Two Dimensional ◽  
Alizarin Red ◽  
Alp Activity ◽  
Growing Cell

Abstract This study aimed to investigate the influence of a three-dimensional cell culture model and bioactive glass (BG) particles on the expression of osteoblastic phenotypes in rat calvaria osteogenic cells culture. Cells were seeded on two-dimensional (2D) and three-dimensional (3D) collagen with BG particles for up to 14 days. Cell viability and alkaline phosphatase (ALP) activity was performed. Cell morphology and immunolabeling of noncollagenous bone matrix proteins were assessed by epifluorescence and confocal microscopy. The expressions of osteogenic markers were analyzed using RT-PCR. Mineralized bone-like nodule formation was visualized by microscopy and calcium content was assessed quantitatively by alizarin red assay. Experimental cultures produced a growing cell viability rate up to 14 days. Although ALP activity at 7 days was higher on BG cultures, cells on 3D and 3D+BG had an activity decrease of ALP at 14 days. Three-dimensional conditions favored the immunolabeling for OPN and BSP and the expression of ALP and COL I mRNAs. BG particles influenced positively the OC and OPN mRNAs expression and calcified nodule formation in vitro. The results indicated that the 3D cultures and BG particles contribute to the expression of osteoblastic phenotype and to differentiated and mineralized matrix formation.

Isolation and culture of wapiti (Cervus elaphus) satellite cells

2000 ◽  
Vol 80 (2) ◽  
pp. 303-309
Author(s):  
N.M. Burton ◽  
L. Shipley ◽  
K. M. Byrne ◽  
J. L. Vierck ◽  
M. V. Dodson
Keyword(s):  
Cell Viability ◽  
Satellite Cells ◽  
Cervus Elaphus ◽  
Horse Serum ◽  
Media Effect ◽  
In Vitro Proliferation ◽  
Pig Skin ◽  
Defined Media ◽  
Proliferation And Differentiation

Myogenic satellite cells (SC) were isolated from the sternomandibularis muscles of two, 227 kg, male wapiti (Cervus elaphus) and studied in primary cell culture. Wapiti-derived SC were capable of attaching to culture substrata and following the myogenic program of proliferation and differentiation to form multinucleated myotubes. Wapiti SC attached equally well to pig skin gelatin (PSG), fibronectin (FN), Matrigel® and plastic (P > 00.05), but cell viability measured at 120 h varied depending on initial substratum type. Pig skin gelatin (0.02% wt vol−1) was chosen for the majority of subsequent experimentation for cost efficiency. The greatest amount of wapiti SC proliferation was observed in media containing 10% (vol/vol−1) horse serum (HS), 15% HS and 15% fetal bovine serum (FBS) (P > 0.05). Wapiti SC proliferated more when exposed to HS and FBS than to sheep serum (SS) (P < 0.05). No proliferation, differentiation or decrease in cell viability was observed in Dulbecco's Modified Eagle Medium (DMEM) + 1% HS, DMEM + 2% HS, DMEM + 3% HS or DMEM + 4% HS (P > 0.05) after 120 h in vitro. Proliferation of SC was doubled when insulin was added to both 10% HS- and 2% HS-containing media (P < 0.05). Although insulin alone in serum-containing media did not promote fusion of wapiti SC, two defined media (ITT and ITT-CF) that contain insulin did promote fusion of wapiti SC cultures. ITT-CF induced 3% fusion of wapiti SC into myotubes, and ITT induced 1% (P < 0.05). There was also an increase in total cell numbers in SC exposed to ITT-CF in comparison with ITT, ovine defined media (ODM) or ovine defined media-Modified (ODM-Mod))(P < 0.05). Although defined media differed in their ability to induce proliferation or differentiation (P < 0.05), the substrata on which the SC were plated did not influence the defined media effect on SC activity (P > 0.05). Satellite cells exposed to ITT and ITT-CF differed morphologically from SC exposed to ODM and ODM-Mod, which may suggest that formulation differences are influencing wapiti-derived SC proliferation and differentiation. Key words: Wapiti, satellite cells, primary culture, myotubes

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