Concentration-dependent effect of fumonisin B1 on apoptosis in oesophageal cancer cells

2017 ◽  
Vol 37 (7) ◽  
pp. 762-771 ◽  
Author(s):  
RB Khan ◽  
A Phulukdaree ◽  
AA Chuturgoon
Keyword(s):  
Cell Viability ◽  
Oesophageal Cancer ◽  
Caspase 3 ◽  
Quantitative Polymerase Chain Reaction ◽  
Fumonisin B1 ◽  
Annexin V ◽  
Tail Length ◽  
Bax Protein ◽  
Genotoxic Carcinogens ◽  
Parp 1

The geographical distribution of oesophageal cancer is linked to the exposure of fumonisin B1 (FB1), a mycotoxin produced by fungi that contaminates staple food worldwide. Non-genotoxic carcinogens like FB1 disturb homeostasis through increased cell proliferation or suppression of apoptosis. This study investigated the involvement of FB1 (0–20 μM) in spindle-shaped N-cadherin (+) CD45 (−) osteoblastic (SNO) cell death. Cell viability and death were assessed using the MTS and Annexin V-Fluos assays, respectively. Caspase activities were determined luminometrically and the comet assay assessed DNA damage. Induction of oxoguanine glycosylase 1 (OGG1) was measured using quantitative Polymerase Chain Reaction (qPCR), while cleaved poly (ADP-ribose) polymerase 1 (PARP-1) and Bax were determined by western blotting. Cell viability and PARP-1 cleavage were not affected by 1.25 μM FB1, but phosphatidylserine externalization, Bax protein expression, caspase activity, comet tail length and OGG1 transcripts were increased. The reduced cell viability in 10 μM FB1-treated cells was accompanied by corresponding increases in externalized phosphatidylserine, Bax, caspase-3/7 activity and cleaved PARP-1. The OGG1 transcripts were not significantly increased, but comet tails were increased. Bax, caspase-3/7 activities and cleaved PARP-1 were inhibited at 20 μM FB1. In addition, the OGG1 transcript levels were decreased ( p < 0.0001) along with comet lengths ( p < 0.0001). This study showed that FB1-induced apoptosis in SNO cells may be caspase-dependent or caspase-independent; the pathway used depends on the exposure concentration.

scholarly journals The Protective Effect of Fluorofenidone against Cyclosporine A-Induced Nephrotoxicity

2019 ◽  
Vol 44 (4) ◽  
pp. 656-668 ◽  
Author(s):  
Yang Chen ◽  
Nasui Wang ◽  
Qiongjing Yuan ◽  
Jiao  Qin ◽  
Gaoyun Hu ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cyclosporine A ◽  
Caspase 3 ◽  
Renal Tubule ◽  
Annexin V ◽  
Type I ◽  
Tubulointerstitial Injury ◽  
Tubule Cells ◽  
Parp 1 ◽  
Tgf Β1

Background/Aims: Cyclosporine A (CsA) is an immunosuppressant drug that is used during organ transplants. However, its utility is limited by its nephrotoxic potential. This study aimed to investigate whether fluorofenidone (AKF-PD) could provide protection against CsA-induced nephrotoxicity. Methods: Eighty-five male Sprague-Dawley rats were divided into 5 groups: drug solvent, CsA, CsA with AKF-PD (250, 500 mg/kg/day), and CsA with pirfenidone (PFD, 250 mg/kg/day). Tubulointerstitial injury index, extracellular matrix (ECM) deposition, expression of type I and IV collagen, transforming growth factor (TGF)-β1, platelet-derived growth factor (PDGF), Fas ligand (FASL), cleaved-caspase-3, cleaved-poly(ADP-ribose) polymerase (PARP)-1, and the number of transferase-mediated nick end-labeling (TUNEL)-positive renal tubule cells were determined. In addition, levels of TGF-β1, FASL, cleaved-caspase-3, cleaved-PARP-1, and number of annexin V-positive cells were determined in rat proximal tubular epithelial cells (NRK-52E) treated with CsA (20 μmol/L), AKF-PD (400 μg/mL), PFD (400 μg/mL), and GW788388 (5 μmol/L). Results: AKF-PD (250, 500 mg/kg/day) significantly reduced tubulointerstitial injury, ECM deposition, expression of type I and IV collagen, TGF-β1, PDGF, FASL, cleaved-caspase-3, cleaved-PARP-1, and number of TUNEL-positive renal tubule cells in the CsA-treated kidneys. In addition, AKF-PD (400 μg/mL) significantly decreased TGF-β1, FASL, cleaved-caspase-3, and PARP-1 expression in NRK-52E cells and further reduced the number of annexin V-positive cells. Conclusion: AKF-PD protect kidney from fibrosis and apoptosis in CsA-induced kidney injury.

Iron-Overload Induces Apoptosis in Cardiomyocytes and Hepatocytes Via Mitochondrial/Caspase-3 Pathways.

Blood ◽  
2008 ◽  
Vol 112 (11) ◽  
pp. 1872-1872
Author(s):  
Mo Yang ◽  
Shing Chan ◽  
Yiu Fai Cheung ◽  
Shau Yin Ha ◽  
Godfrey ChiFung Chan
Keyword(s):  
Iron Overload ◽  
Protective Effect ◽  
Cell Viability ◽  
Caspase 3 ◽  
Annexin V ◽  
Apoptotic Cells ◽  
H9c2 Cells ◽  
Plot Analysis ◽  
Iron Treatment ◽  
Induced Apoptosis

Abstract Cardiomyopathy and liver damage due to iron-overload are the major complications in patients with beta-thalassaemia major. Iron-overload may induce apoptosis in cardiomyocytes and hepatic cells, and that TPO may exert protective effect on apoptosis of cardiomyocytes (Circulation, 2006). In this study, we demonstrated firstly that iron induced apoptosis in cardiomyocytes. Using H9C2 cells, we have shown that iron reduced cell viability in a dose-dependent manner (0.003–3 mM) (n=6). By annexin V and PI staining, apoptotic cells were found to be significantly increased after iron treatment (0.3 mM, 72 hrs) (n=6). The expression of active caspase-3 was significantly increased in iron-treated cells. Furthermore, iron treatment increased the proportion of cells containing JC-1 monomers, indicating a trend in the drop of mitochondrial membrane potential (n=6). Secondly, we found that TPO exerted cardio-protective effect on iron-induced apoptosis. H9C2 cells were cultured in the presence of iron (0.3 mM) with or without TPO (5, 10, 20, 50, 100 ng/mL, 72 hrs). The cell viability was significantly increased with the treatment of TPO at 50 ng/mL and 100 ng/mL (n=4). Dot-plot analysis of annexin V/PI staining demonstrated that TPO (50 ng/mL) significantly reduced the population of apoptotic cells (n=6). Incubation with TPO also decreased the iron-induced caspase-3 expression (n=6). Flow cytometric dot-plot analysis of H9C2 cells also showed trends of amelioration of the increase in JC-1 monomers in the iron plus TPO group (n=6). The population of phospho-Akt and Erk1/2 were also significantly increased after treatment by TPO (P&lt;0.05, n=4). Human liver cell line MIHA was also used as a cell model. We showed that iron-overload reduced cell viability in a dose-dependent manner (0.0375–0.6 mM) (n=7). By annexin V and PI staining, apoptotic cells were found to be significantly increased after iron treatment (0.15–0.6 mM) for 72 hrs (n=7). The expression of active caspase-3 was also significantly increased in iron-treated cells (n=5). We also found that TPO exerted proliferation effect on MIHA cell by activation of phospho-Akt. However, MIHA cells were cultured in the presence of iron (0.3 mM) with TPO (50 ng/mL, 72 hrs). The cell viability was not significantly increased with the treatment of TPO (n=5). Dot-plot analysis of annexin V/PI staining did not demonstrated that TPO reduced the population of apoptotic cells induced by iron-overload (n=5). Also, incubation with TPO did not decrease the iron-induced caspase-3 expression in these cells (n=5). Our findings suggest that iron-overload induces apoptosis in cardiomyocytes and hepatocytes via mitochondrial/caspase-3 pathways and that TPO might exert a protective effect on iron-overload induced apoptosis via the activation of Akt and Erk1/2 pathways in cardiomyocytes.

Bone Marrow Stromal Cell-Derived Exosomes Facilitate Multiple Myeloma Cell Survival Through Inhibition Of The JNK Pathway

Blood ◽  
2013 ◽  
Vol 122 (21) ◽  
pp. 679-679
Author(s):  
Jinheng Wang ◽  
An Hendrix ◽  
Elke De Bruyne ◽  
Els Van Valckenborgh ◽  
Eddy Himpe ◽  
...  
Keyword(s):  
Multiple Myeloma ◽  
Bone Marrow ◽  
Cell Proliferation ◽  
Western Blot ◽  
Cell Viability ◽  
Cell Survival ◽  
Caspase 3 ◽  
Annexin V ◽  
Jnk Pathway ◽  
Functional Components

Abstract Interplay between bone marrow stromal cells (BMSCs) and multiple myeloma (MM) cells plays a crucial role in MM pathogenesis by secreting growth factors, cytokines, and other functional components. Exosomes are 30-100nm diameter membranous vesicles constitutively released by several cell types including reticulocytes, cytotoxic T lymphocytes, B lymphocytes, epithelial and endothelial cells. Exosomes mediate local cell-cell communication by transferring mRNAs, miRNAs and proteins. Due to their ability to transfer functional components, exosomes play multiple roles by stimulating target cells, transferring membrane receptors, delivering proteins, and inducing epigenetic changes in recipient cells. Although the promotion of MM growth and survival induced by BMSCs has been studied, the role of BMSC-derived exosomes in this action remains unclear. Here, we investigated the effect and mechanisms of BMSC-derived exosomes on the proliferation and survival of MM cells using the murine 5T33MM model. This model mimics the human disease closely and of this model two lines exist: the 5T33MMvv model which is propagated in vivo and the 5T33MMvt line which is derived from 5T33MMvv cells but which can grow stroma-independently. Exosomes were isolated from conditioned medium using the ExoQuick-TC Exosome Precipitation Solution (System Biosciences) after culture of primary BMSCs obtained from naïve C57BL/KaLwRij mice or 5T33MM diseased mice. The size of exosomes derived from naïve BMSCs, 5T33 BMSCs and 5T33MMvt cells were confirmed using a NanoSight LM10. Several exosomal markers such as CD63, Flotillin-1, heat shock protein 90 (HSP90), and HSP70 were detected using Western blot. We co-cultured the BMSCs or MM cells with fluorescent dye-labeled exosomes to examine whether exosomes could be transferred into cells. The results showed that both naïve and 5T33 BMSC-derived exosomes could fuse with 5T33MMvt cells and that the uptake of 5T33MMvt cell-derived exosomes by BMSCs was also observed. As several cytokines were found to be present in BMSC- and MMvt cell-derived exosomes, this suggests that BMSCs and MM cells could exchange cytokines with each other through exosomes secretion and uptake. Furthermore, the cytokine composition of 5T33BMSC-derived exosomes compared to naïve BMSC-derived exosomes was different. We next performed luminescent cell viability assays, BrdU cell proliferation assays and 7-AAD/annexin-V stainings to examine the effects of BMSC-derived exosomes on MM cell viability, proliferation and survival, respectively. Both naïve and 5T33 BMSC-derived exosomes increased 5T33MMvt and MMvv cell viability in a dose- and time-dependently manner. BrdU uptake in 5T33MMvt and MMvv cells was also increased after treatment with BMSC-derived exosomes. Significantly reduced apoptosis of 5T33 MMvt and MMvv cells was observed when they were treated with BMSC-derived exosomes as judged by 7-AAD/annexin-V staining. 5T33MMvt and MMvv cells were treated with different amounts of BMSC-derived exosomes and apoptosis-related proteins Bcl-2, Bax, and caspase-3 were determined using western blot. Bcl-2 was increased slightly and activated (cleaved) caspase-3 was reduced after co-culture with exosomes, coinciding with the results of 7-AAD/annexin-V staining. To elucidate the mechanisms responsible for exosome-induced MM cell survival, we examined the activation of several proteins involved. Reduced phosphorylation of p53, p38MAPK and JNK were detected when 5T33MMvt were treated with naïve-BMSC-derived exosomes for 24h, whereas phosphorylated Erk1/2, Akt, and IGF1Rβ were not changed. Surprisingly, activation of p53 and p38MAPK were not changed after the treatment with 5T33 BMSC-derived exosomes. 5T33 BMSC-derived exosomes further decreased the activation of JNK, Bim expression and phosphorylated Bim compared to naïve BMSC-derived exosomes. As Bim is a pro-apoptosis protein, mainly regulated by the JNK pathway; promotion of MM cell survival likely results from the inhibition of the JNK pathway by BMSC-derived exosomes. In summary, our results demonstrate a positive role for BMSC-derived exosomes in induction of MM cell proliferation and survival. BMSC-derived exosomes could inhibit the JNK pathway, thereby reducing caspase-3 activation and protecting MM cells from apoptosis. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Antioxidant effects on hypoxia-induced oxidative stress and apoptosis in rat rotator cuff fibroblasts

2021 ◽  
Vol 41 ◽  
pp. 680-693
Author(s):  
RJ Kim ◽  
◽  
SH An ◽  
JY Gwark ◽  
HB Park
Keyword(s):  
Rotator Cuff ◽  
Cell Viability ◽  
Caspase 3 ◽  
Hypoxia Inducible Factor ◽  
Matrix Metalloproteinase 2 ◽  
Heme Oxygenase 1 ◽  
Ros Production ◽  
Hypoxia Group ◽  
Pre Treatment ◽  
Parp 1

Most cells, highly sensitive to oxygen levels, undergo apoptosis under hypoxia. Therefore, the involvement of hypoxia in rotator cuff tendon degeneration has been proposed. While previous studies have reported that hypoxia induces apoptosis in rotator cuff fibroblasts (RCFs), little research has investigated whether antioxidants have cytoprotective effects against RCF apoptosis. The present study aimed at determining whether the antioxidant N-acetylcysteine (NAC) exerted cytoprotective effects against hypoxia-induced RCF apoptosis. Third-passage rat RCFs were divided into normoxia, NAC, hypoxia and NAC-hypoxia groups. The hypoxia inducer was 1,000 µmol/L cobalt chloride (CoCl2); the antioxidant was 20 mmol/L NAC. Expressions of hypoxia-inducible factor-1α (HIF-1α) and heme oxygenase-1 (HO-1), cell viability, intracellular reactive oxygen species (ROS) production, apoptosis rates as well as expressions of cleaved caspase-3, cleaved poly ADP-ribose polymerase-1 (PARP-1), vascular endothelial growth factors-β (VEGF-β) and matrix metalloproteinase-2 (MMP-2) were evaluated. Expression of HIF-1α and HO-1 was significantly higher in the hypoxia group than in the normoxia group (p < 0.001). Cell viability was significantly lower in the hypoxia group than in the normoxia group (p < 0.001). Intracellular ROS production, apoptosis rate and expressions of cleaved caspase-3, cleaved PARP-1, VEGF-β and MMP-2 were significantly higher in the hypoxia group than in the normoxia group (p < 0.001). All these responses were significantly attenuated by pre-treatment with NAC (p ≤ 0.001). ROS were involved in hypoxic RCF apoptosis induced by CoCl2; NAC, an ROS scavenger, inhibited hypoxia-induced RCF apoptosis by inhibiting ROS production.

The Anti-Inflammatory Investigational Agent LMP-420 Demonstrates in Vitro Cytotoxic Activity against Chronic Lymphocytic Leukemia Cells.

Blood ◽  
2009 ◽  
Vol 114 (22) ◽  
pp. 2358-2358
Author(s):  
Daphne R Friedman ◽  
Yvonne Mowery ◽  
Margaret Kennedy ◽  
Karen M Bond ◽  
J. Brice Weinberg ◽  
...  
Keyword(s):  
High Risk ◽  
Chronic Lymphocytic Leukemia ◽  
Cell Viability ◽  
Cytotoxic Activity ◽  
Colony Formation ◽  
Cytotoxic Effect ◽  
Caspase 3 ◽  
Annexin V ◽  
Lymphocytic Leukemia ◽  
Serial Dilutions

Abstract Abstract 2358 Poster Board II-335 Background: Chronic Lymphocytic Leukemia (CLL) is a common incurable hematologic malignancy. When therapy is required, maximizing durable responses is often at the risk of increasing toxicity. Thus, developing novel therapeutic agents that have minimal overlapping toxicity with currently used chemotherapy would be advantageous. To this end, we investigated LMP-420, a boronic acid containing purine nucleoside analogue, that potently inhibits tumor necrosis factor alpha (TNF) transcription in stimulated peripheral blood mononuclear cells (PBMCs) without affecting cell viability. Since TNF has been implicated in promoting CLL cell viability and can be produced by CLL cells themselves, we hypothesized that LMP-420 would be cytotoxic for CLL lymphocytes, either alone or in combination with fludarabine. Methods: To test the activity of LMP-420, we negatively selected circulating CLL cells from blood collected from patients using the RosetteSep B-cell enrichment cocktail (StemCell Technologies) and a Ficoll-Hypaque gradient. This method yields greater than 95% purity of malignant lymphocytes, determined by immunophenotyping CD5+CD19+ cells. Prognostic markers such as IgVH mutation status, CD38 and ZAP70 expression, and interphase cytogenetics were determined as previously described. We assessed the fractional toxicity and 50% effective dose (ED50) of LMP-420, fludarabine, or the combination, with the MTS colorimetric cytotoxicity assay, in which CLL cells were incubated for three days in Hybridoma media + 10% fetal bovine serum with serial dilutions of drug alone or in combination. Apoptosis was measured via annexin V flow cytometry methods and caspase 3/7 activity assays. We determined the effect of LMP-420 compared to fludarabine on the normal hematopoietic system by testing serial dilutions of both agents in killing normal PBMCs and in suppressing erythroid and myeloid colony formation. Results: The median ED50 of LMP-420 for CLL cells was 423 nM (range 0.01 to 2224 nM, n = 21). Two patients had high-risk cytogenetics (17p or 11q deletions), and their ED50 values for LMP-420 were 691 and 90 nM, respectively. The cytotoxic effect of fludarabine was potentiated on average 80 or 261 fold with the addition of LMP-420 at concentrations of 62 or 250 nM, respectively (ranges 1.14 – 947 and 1.19 – 2754). This agent killed malignant lymphocytes by apoptotic mechanisms in a dose-responsive fashion, as demonstrated by both Annexin V staining and caspase 3/7 activity assays. While LMP-420 has potent anti-CLL activity, it has minimal effects on normal hematologic cells. For example, fludarabine suppresses erythroid and myeloid colony formation by greater than 50% at a concentration of 1 uM, while this level of inhibition is seen for LMP-420 at a concentration of 90 uM. The average cytotoxic ED50 of LMP-420 on normal PBMCs using the MTS assay was greater than 90 uM, whereas for fludarabine, it was 5.3 uM. This finding was confirmed with apoptosis assays. Conclusions: The results of these experiments demonstrate that LMP-420, a novel inhibitor of TNF expression, has cytotoxic activity against CLL cells, including those with high-risk features. LMP-420 appears to increase the cytotoxic effect of the chemotherapy agent fludarabine, while imparting minimal increase in hematologic toxicity. Thus, LMP-420 is promising new therapeutic agent in CLL. Disclosures: No relevant conflicts of interest to declare.

The p55 TNF receptor mediates TNF inhibition of osteoblast differentiation independently of apoptosis

2005 ◽  
Vol 288 (5) ◽  
pp. E1011-E1018 ◽  
Author(s):  
Linda C. Gilbert ◽  
Janet Rubin ◽  
Mark S. Nanes
Keyword(s):  
Cell Viability ◽  
Caspase 3 ◽  
Annexin V ◽  
Inhibitory Effect ◽  
Tnf Receptor ◽  
Trypan Blue Exclusion ◽  
Total Dna ◽  
Factor Α ◽  
Tnf Inhibition

After menopause, increased tumor necrosis factor-α (TNF-α) stimulates bone resorption while inhibiting differentiation of new bone-forming osteoblasts (OB). TNF receptors, p55 and p75, signal similar intracellular pathways, but only p55 activates apoptosis. To evaluate the relationship between the TNF receptor mediating inhibition of OB differentiation and the role of apoptosis, marrow stromal cells (MSC) were cultured from mice deficient in either or both receptors. Cells grown in ascorbate and β-glycerophosphate produce alkaline phosphatase and osteocalcin and mineralize matrix. Treatment of wild-type or p55+/+/p75−/− MSC with murine TNF (binds p55 and p75) or human TNF (binds only p55) inhibited OB differentiation. TNF did not inhibit OB differentiation in p55−/− MSC. Expression of p75 modestly attenuated sensitivity to TNF. To determine the role of apoptosis, changes in total DNA, cell viability, caspase 3, and percentage of annexin V-positive cells were measured in MSC and preosteoblastic MC3T3 cells. TNF treatment that reduced differentiation by 50% did not decrease cell viability or increase apoptosis, as determined by alamar blue reduction, trypan blue exclusion, and percentage of annexin V-positive cells. TNF increased caspase 3 activity 1.5-fold in MC3T3 and insignificantly in MSC cells compared with >4-fold after 4 h actinomycin D. Treatment of MSC or MC3T3 cells with three caspase inhibitors failed to reverse the inhibitory effect of TNF on OB differentiation despite inhibition of caspase activity. These results suggest that the p55 receptor is essential, and p75 dispensable, for TNF inhibition of OB differentiation through a mechanism that does not require apoptosis.

scholarly journals Protective effect of resveratrol against corticosterone-induced neurotoxicity in PC12 cells

2019 ◽  
Vol 10 (1) ◽  
pp. 235-240 ◽  
Author(s):  
Ye Zhang ◽  
Yun He ◽  
Ning Deng ◽  
Yan Chen ◽  
Jiecong Huang ◽  
...  
Keyword(s):  
Protein Expression ◽  
Protective Effect ◽  
Cell Viability ◽  
Pc12 Cells ◽  
Caspase 3 ◽  
Annexin V ◽  
Cell Counting ◽  
Depressant Effect ◽  
Protective Efficiency ◽  
Related Proteins

Abstract Objective Resveratrol(RES) is a natural polyphenol which possesses an anti-depressant effect. However, the mechanisms of its anti-depressant effect remain unclear. The aim of the study is to investigate the potential mechanisms in the neuro-protective efficiency in the corticosterone-induced pheochromacytoma 12 (PC12) cells. Methods PC12 cells were treated with 200 μM of corticosterone in the absence or presence of different concentrations of RES for 24 h. Then, cell viability was measured by Cell Counting Kit-8 assay. Apoptosis of PC12 cells was measured by Annexin V-FITC and Propidium iodide (PI) labelling. The expression of apoptosis-related proteins including Bax, Bcl-2, caspase-3 was determined by western blotting. Results The results showed that treatment with 200 μM of corticosterone induced cytotoxicity in PC12 cells. However, different concentrations of RES (2.5μmol/L, 5μmol/L and 10 μmol/L) significantly increased the cell viability, suppressed the apoptosis of PC12 cells, down-regulated Bax and caspase-3 protein expression, and up-regulated Bcl-2 protein expression, compared to the model group (p<0.05). Conclusion Resveratrol has a protective effect on corticosterone-induced neurotoxicity in PC12 cells, which may be related to the apoptosis via inhibition of apoptosis-related proteins and displays the antidepressant-like effect.

scholarly journals Mitigation of H2O2-Induced Mitochondrial-Mediated Apoptosis in NG108-15 Cells by Novel Mesuagenin C fromMesua kunstleri(King) Kosterm

2012 ◽  
Vol 2012 ◽  
pp. 1-18 ◽  
Author(s):  
Gomathi Chan ◽  
Muhamad Noor Alfarizal Kamarudin ◽  
Daniel Zin Hua Wong ◽  
Nor Hadiani Ismail ◽  
Faizuri Abdul Latif ◽  
...  
Keyword(s):  
Cell Viability ◽  
Caspase 3 ◽  
Chromatin Condensation ◽  
Annexin V ◽  
Data Interpretation ◽  
Hexane Extract ◽  
Neuroprotective Activity ◽  
Apoptotic Pathway ◽  
Chemical Structures ◽  
Induced Apoptosis

This study was aimed to isolate and evaluate neuroprotective compounds from the hexane extract of the bark ofMesua kunstleri(Clusiaceae) on H2O2-induced apoptosis in NG108-15 cells. Five 4-phenylcoumarins were isolated by using various chromatographic techniques via neuroprotective activity-guided fractionation and isolation from the active hexane extract. The chemical structures of the isolated compounds were confirmed by NMR spectroscopic data interpretation and comparison with literature values. Cell viability data demonstrated that mesuagenin C3significantly increased cell viability. Hoechst 33342/PI staining illustrated mesuagenin C3was able to abate the nuclear shrinkage, chromatin condensation and formation of apoptotic bodies. Pretreatment with mesuagenin C3reduced total annexin V positive cells and increased the level of intracellular glutathione (GSH). Mesuagenin C3attenuated membrane potential (Δψm), reduced Bax/Bcl-2 ratio and inactivated of caspase-3/7 and -9. These results indicated that mesuagenin C3could protect NG108-15 cells against H2O2-induced apoptosis by increasing intracellular GSH level, aggrandizingΔψm, and modulating apoptotic signalling pathway through Bcl-2 family and caspase-3/7 and -9. These findings confirmed the involvement of intrinsic apoptotic pathway in H2O2-induced apoptosis and suggested that mesuagenin C3may have potential therapeutic properties for neurodegenerative diseases.

scholarly journals Zinc attenuates ecstasy-induced apoptosis through downregulation of caspase-3 in cultured TM3 cells: An experimental study

2020 ◽  
Author(s):  
Marziyeh Lozeie ◽  
Morteza Bagheri ◽  
Isa Abdi Rad ◽  
Nadia Hossein-Zadeh ◽  
Mahdyieh Nasir-Zadeh
Keyword(s):  
Cell Viability ◽  
Caspase 3 ◽  
Quantitative Polymerase Chain Reaction ◽  
Mrna Level ◽  
Control Cell ◽  
Control Group ◽  
High Concentration ◽  
Group I ◽  
The Mean ◽  
Induced Apoptosis

Background: 3, 4-Methylenedioxymethamphetamine (MDMA) is commonly known as the most famous amphetamine derivative. Objective: To evaluate the influence of zinc on MDMA-induced apoptosis and caspase- 3 gene expression in Leydig cell line (TM3). Materials and Methods: Leydig cells were studied in differenet treatment groups regarding MDMA (0, 0.5, 1, 3, 5 mM) and zinc (0, 4, 8, 16, 32 μM). By the way, the effective concentration was determined to be 5 mM for MDMA and 8 μM for zinc. Then, TM3 cells were cultured in free medium as control (group I), medium containing MDMA (5 mM) (group II), zinc (8 μM) (group III), and zinc (8 μM) prior to MDMA (5 mM) (group IV) as well as in an untreated group (control). Cell viability was assessed at different times after cell culture by MTT assay. The mRNA expression level of caspase-3 was analyzed using real-time quantitative polymerase chain reaction. Results: The cellular viability was significantly reduced in TM3 cells after 24 hr and 48 hr exposure time regarding different concentrations of MDMA as well as high concentration of zinc (16 and 32 μM). Cell viability was increased in the group that received zinc (8 μM) before addition of MDMA (5 mM) compared to the control and MDMA groups. The mean ± SE of fold was 22.40 ± 7.5, 0.06 ± 0.02, and 0.009 ± 0.003 in MDMA, zinc, and zinc + MDMA groups, respectively. The mean of caspase-3 mRNA level was significantly increased in the MDMA-treated group (5 mM), while the relative expression of caspase-3 gene was significantly decreased in the zinc (8 μM) + MDMA (5 mM) group compared with the MDMA (5 mM) group (p = 0.001). Conclusion: Dietary intake of zinc has a protective effect against MDMA consumption in mouse. Key words: Zinc, MDMA, Apoptosis, TM3 cells.

Protective effect of dexmedetomidine on neuronal hypoxic injury through inhibition of miR-134

2021 ◽  
pp. 096032712110237
Author(s):  
Y-J Li ◽  
D-Z Zhang ◽  
Y Xi ◽  
C-A Wu
Keyword(s):  
Oxidative Stress ◽  
Cell Viability ◽  
Pc12 Cells ◽  
Cell Apoptosis ◽  
Caspase 3 ◽  
Cell Damage ◽  
Annexin V ◽  
Protective Effects ◽  
Gene And Protein Expression ◽  
And Oxidative Stress

Objective: To explore the mechanism of dexmedetomidine (DEX)-mediated miR-134 inhibition in hypoxia-induced damage in PC12 cells. Methods: Hydrogen peroxide (H2O2)-stimulated PC12 cells were divided into control, H2O2, DEX + H2O2, miR-NC/inhibitor + H2O2, and miR-NC/ mimic + DEX + H2O2 groups. Cell viability and apoptosis were assessed by the 3-(4,5-dimethylthiazol(-2-y1)-2,5-diphenytetrazolium bromide (MTT) assay and Annexin V-FITC/PI staining, while gene and protein expression levels were detected by qRT-PCR and western blotting. Reactive oxygen species (ROS) levels were tested by 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA) staining, and malondialdehyde (MDA) content was determined with a detection kit. Results: DEX treatment decreased H2O2-elevated miR-134 expression. H2O2-induced PC12 cell damage was improved by DEX and miR-134 inhibitor; additionally, cell viability was increased, while cell apoptosis was reduced. In addition, both DEX and miR-134 inhibitor reduced the upregulated expression of cleaved caspase-3 and increased the downregulated expression of Bcl-2 in H2O2-induced PC12 cells. However, compared to that in the DEX + H2O2 group, cell viability in the mimic + DEX + H2O2 group was decreased, and the apoptotic rate was elevated with increased cleaved caspase-3 and decreased Bcl-2 expression. Inflammation and oxidative stress were increased in H2O2-induced PC12 cells but improved with DEX or miR-134 inhibitor treatment. However, this improvement of H2O2-induced inflammation and oxidative stress induced by DEX in PC12 cells could be reversed by the miR-134 mimic. Conclusion: DEX exerts protective effects to promote viability and reduce cell apoptosis, inflammation, and oxidative stress in H2O2-induced PC12 cells by inhibiting the expression of miR-134.

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