scholarly journals Protection of SH-SY5Y Neuronal Cells from Glutamate-Induced Apoptosis by 3,6′-Disinapoyl Sucrose, a Bioactive Compound Isolated from Radix Polygala

2012 ◽  
Vol 2012 ◽  
pp. 1-5 ◽  
Author(s):  
Yuan Hu ◽  
Jie Li ◽  
Ping Liu ◽  
Xu Chen ◽  
Dai-Hong Guo ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Mtt Assay ◽  
Neuronal Cells ◽  
Bioactive Compound ◽  
Glutamate Toxicity ◽  
Flow Cytometry Assay ◽  
Neuroprotective Effects ◽  
Induced Apoptosis ◽  
Proapoptotic Gene

The neuroprotective effects of 3,6′-disinapoyl sucrose (DISS) from Radix Polygala against glutamate-induced SH-SY5Y neuronal cells injury were evaluated in the present study. SH-SY5Y neuronal cells were pretreated with glutamate (8 mM) for 30 min followed by cotreatment with DISS for 12 h. Cell viability was determined by (3,4,5-dimethylthiazol-2-yl)-2,5-diphenylte-trazolium bromide (MTT) assay, and apoptosis was confirmed by cell morphology and flow cytometry assay, evaluated with propidium iodide dye. Treatment with DISS (0.6, 6, and 60 μmol/L) increased cell viability dose dependently, inhibited LDH release, and attenuated apoptosis. The mechanisms by which DISS protected neuron cells from glutamate-induced excitotoxicity included the downregulation of proapoptotic gene Bax and the upregulation of antiapoptotic gene Bcl-2. The present findings indicated that DISS exerts neuroprotective effects against glutamate toxicity, which might be of importance and contribute to its clinical efficacy for the treatment of neurodegenerative diseases.

scholarly journals Evaluation of the Anti-apoptotic and Anti-cytotoxic Effect of Epicatechin Gallate and Edaravone on SH-SY5Y Neuroblastoma Cells

2019 ◽  
pp. 619-630
Author(s):  
Mohammad Shokrzadeh ◽  
Hashem Javanmard ◽  
Golpar Golmohammad Zadeh ◽  
Hossein Asgarian Omran ◽  
Mona Modanlou ◽  
...  
Keyword(s):  
Synergistic Effect ◽  
Cell Viability ◽  
Mtt Assay ◽  
Neuroprotective Effect ◽  
Neuroblastoma Cells ◽  
Protective Effects ◽  
Older Individuals ◽  
Epicatechin Gallate ◽  
Neuroprotective Effects ◽  
Late Apoptosis

Introduction: Parkinson disease (PD) is the second most common neurodegenerative disease affecting older individuals with signs of motor disability and cognitive impairment. Epicatechin (EC) and edaravone have neuroprotective effects most probably due to their antioxidant activity; however, a limited number of studies have considered their role in PD. This research aimed at investigating the neuroprotective effect of EC and edaravone in a neurotoxin-induced model of PD. Methods: An in vitro model of PD was made by subjecting SH-SY5Y neuroblastoma cells to neurotoxin: 6-hydroxydopamine (6-OHDA) 100 µM/well. The cytoprotective effect of EC and edaravone in five concentrations on cell viability was tested using the MTT assay. The apoptotic assay was done by annexin V and propidium iodide method using flow cytometry. Results: According to the MTT assay analysis, EC and edaravone had protective effects against 6-OH DA-induced cytotoxicity in SH-SY5Y neuroblastoma cells that were much more significant for edaravone and also a relative synergistic effect between EC and edaravone was observed. The apoptotic analysis showed that edaravone alone could decrease early and late apoptosis, whereas EC diminished early apoptosis, but enhanced late apoptosis and necrosis. Besides, co-treatment of edaravone and EC had a synergistic effect on decreasing apoptosis and increasing cell viability. Conclusion: The protective effect of edaravone on apoptosis and cytotoxicity was demonstrated clearly and EC had a synergistic effect with edaravone.

scholarly journals Neuroprotective Effects of 7-Geranyloxycinnamic Acid from Melicope lunu ankenda Leaves

Molecules ◽  
2020 ◽  
Vol 25 (16) ◽  
pp. 3724
Author(s):  
Zeinab Abdulwanis Mohamed ◽  
Enas Mohamed Eliaser ◽  
Mohammed Sani Jaafaru ◽  
Norshariza Nordin ◽  
Costas Ioannides ◽  
...  
Keyword(s):  
Neuronal Cells ◽  
Cytoplasmic Inclusion ◽  
Annexin V ◽  
Neuroprotective Activity ◽  
Neuroprotective Effects ◽  
Human Neuroblastoma ◽  
Differentiated Cells ◽  
Pre Treatment ◽  
Induced Apoptosis

Neurodegenerative diseases (NDDs) are chronic conditions that have drawn robust interest from the scientific community. Phytotherapeutic agents are becoming an important source of chemicals for the treatment and management of NDDs. Various secondary metabolites have been isolated from Melicope lunu-ankenda plant leaves, including phenolic acid derivatives. However, their neuroprotective activity remains unclear. Thus, the aim of this study is to elucidate the in vitro neuroprotective activity of 7-geranyloxycinnamic acid isolated from Melicope lunu-ankenda leaves. The neuroprotective activity was evaluated in differentiated human neuroblastoma (SH-SY5Y) cells by monitoring cell viability using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Moreover, the potential to impair apoptosis in differentiated cells was investigated employing the Annexin V-FITC assay, acridine orange and propidium iodide (AO/PI) staining, and fluorescence microscopy. Morphological assessment and ultrastructural analysis were performed using scanning and transmission electron microscopy to evaluate the effect of 7-geranyloxycinnamic acid on surface morphology and internal features of the differentiated cells. Pre-treatment of neuronal cells with 7-geranyloxycinnamic acid significantly protected the differentiated SH-SY5Y cells against H2O2-induced apoptosis. Cytoskeleton and cytoplasmic inclusion were similarly protected by the 7-geranyloxycinnamic acid treatment. The present findings demonstrate the neuroprotective potential of 7-geranyloxycinnamic acid against H2O2-induced neurotoxicity in neuronal cells, which is an established hallmark of neuronal disorders.

scholarly journals Hexarelin Modulation of MAPK and PI3K/Akt Pathways in Neuro-2A Cells Inhibits Hydrogen Peroxide—Induced Apoptotic Toxicity

2021 ◽  
Vol 14 (5) ◽  
pp. 444
Author(s):  
Ramona Meanti ◽  
Laura Rizzi ◽  
Elena Bresciani ◽  
Laura Molteni ◽  
Vittorio Locatelli ◽  
...  
Keyword(s):  
Hydrogen Peroxide ◽  
Cell Viability ◽  
Caspase 3 ◽  
Mrna Levels ◽  
Protective Effects ◽  
Neuroprotective Effects ◽  
Caspase 7 ◽  
Induced Apoptosis

Hexarelin, a synthetic hexapeptide, exerts cyto-protective effects at the mitochondrial level in cardiac and skeletal muscles, both in vitro and in vivo, may also have important neuroprotective bioactivities. This study examined the inhibitory effects of hexarelin on hydrogen peroxide (H2O2)-induced apoptosis in Neuro-2A cells. Neuro-2A cells were treated for 24 h with various concentrations of H2O2 or with the combination of H2O2 and hexarelin following which cell viability and nitrite (NO2−) release were measured. Cell morphology was also documented throughout and changes arising were quantified using Image J skeleton and fractal analysis procedures. Apoptotic responses were evaluated by Real-Time PCR (caspase-3, caspase-7, Bax, and Bcl-2 mRNA levels) and Western Blot (cleaved caspase-3, cleaved caspase-7, MAPK, and Akt). Our results indicate that hexarelin effectively antagonized H2O2-induced damage to Neuro-2A cells thereby (i) improving cell viability, (ii) reducing NO2− release and (iii) restoring normal morphologies. Hexarelin treatment also reduced mRNA levels of caspase-3 and its activation, and modulated mRNA levels of the BCL-2 family. Moreover, hexarelin inhibited MAPKs phosphorylation and increased p-Akt protein expression. In conclusion, our results demonstrate neuroprotective and anti-apoptotic effects of hexarelin, suggesting that new analogues could be developed for their neuroprotective effects.

scholarly journals Neuroprotective Effects of Coreopsis lanceolata Flower Extract against Oxidative Stress-Induced Apoptosis in Neuronal Cells and Mice

Antioxidants ◽  
2021 ◽  
Vol 10 (6) ◽  
pp. 951
Author(s):  
Hyung Don Kim ◽  
Ji Yeon Lee ◽  
Jeong-Yong Park ◽  
Dong Hwi Kim ◽  
Min Hye Kang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Enzymes ◽  
Pc12 Cells ◽  
Caspase 3 ◽  
Antioxidant Activities ◽  
Neuronal Cells ◽  
Water Extract ◽  
Protective Effects ◽  
Neuroprotective Effects ◽  
Induced Apoptosis

(1) Background: Coreopsis lanceolata L. is a perennial plant of the family Asteraceae, and its flower is known to contain flavonoids with various bioactivities. We evaluated the effect of Coreopsis lanceolata L. flower (CLF) extracts on H2O2-induced oxidative stress (OS) in neuronal cells and mouse neurons. (2) Methods: The flowering part of CL was used as CLF1 (70% ethanol extract) and CLF2 (water extract), and 10 types of phenolic compounds were quantified using high-performance liquid chromatography. To evaluate the neuroprotective effects of CLF, the antioxidant activities of the extracts were measured, and the expression levels of antioxidant enzymes and proteins related to OS-induced apoptosis in neuronal cells and mouse neurons treated with the extracts were investigated. (3) Results: In the in vitro study, CLF ameliorated H2O2-induced oxidative stress and induced the expression of antioxidant enzymes in PC12 cells. Furthermore, CLF1 enhanced the expression of the Bcl-xL protein but reduced the expression of Bax and the cleavage of caspase-3. In the same manner, CLF1 showed neuroprotective effects against OS in vivo. Pretreatment with CLF1 (200 mg/kg) increased the Bcl-2 protein and decreased Bax compared with the 1-methyl-4-phenylpyridinium ion (MPP+)-treated C57BL/6 mice model group. Our results suggest that the protective effects of CLF1 on MPP+-induced apoptosis may be due to its anti-apoptotic activity, through regulating the expression of the Bcl-2 family. (4) Conclusions: CLF1 exerts neuroprotective effects against OS-induced apoptosis in PC12 cells in a Parkinson’s disease model mouse. This effect may be attributable to the upregulation of Bcl-2 protein expression, downregulation of Bax expression, and inhibition of caspase-3 activation. These data indicate that CLF may provide therapeutic value for the treatment of progressive neurodegenerative diseases.

Does Phenytoin Have Neuroprotective Role and Affect Biocompatibility of Decellularized Sciatic Nerve Scaffold?

2020 ◽  
Vol In Press (In Press) ◽  
Author(s):  
Somayyeh Abbaszadeh ◽  
Asadollah Asadi ◽  
Saber Zahri ◽  
Arash Abdolmaleki ◽  
Fariba Mahmoudi
Keyword(s):  
Stem Cells ◽  
Flow Cytometry ◽  
Sciatic Nerve ◽  
Cell Viability ◽  
Mtt Assay ◽  
Good Condition ◽  
Definitive Treatment ◽  
Control Group ◽  
Flow Cytometry Analysis ◽  
Differentiation Potential

Background: Peripheral nervous system injuries are common and currently have no definitive treatment method. Phenytoin is one of the main antiepileptic drugs. Some studies have described a cerebroprotective effect of phenytoin in an established model of global cerebral ischemia. Objectives: In this study, the neuroprotective effects of phenytoin were evaluated on the cultivation and maintenance of Wharton’s jelly stem cells (WJSCs) on acellularized sciatic nerve scaffolds. Methods: In this study, acellular scaffolds from the rat sciatic nerve were prepared by the sondell method. After extraction of cells of MSCs, flow cytometry analysis was executed. Also, cell differentiation potential was evaluated by placement in osteogenic and adipogenic differentiation media for 21 days. Biocompatibility of the scaffold and cell viability were investigated using the MTT assay. The morphological and cell adhesion characteristics of MSCs on acellular scaffolds were compared using SEM micrographs images. Data were analyzed using the one-way analysis of variance (ANOVA) and Tukey post hoc test by SPSS (version 19.0) software. Results: The removal of cells from the scaffold was confirmed by stanning with hematoxylin-eosin, van Gieson's picro-fuchsin, and DAPI. With the aid of flow cytometry analysis and differentiation into bone and fat cells, it was confirmed that extracted cells were mesenchymal stem cells. The results of the MTT assay showed that phenytoin increased cell viability and retention on the scaffold. Conclusions: The study indicated that phenytoin improves the viability of cells and provided a good condition for the growth, survival, and attachment of cells to the scaffold when compared to the control group. These results suggest that phenytoin can be considered a new treatment for nerve regeneration and tissue engineering applications.

Berbamine-Induced Apoptosis in Human Leukemia K562 Cells.

Blood ◽  
2004 ◽  
Vol 104 (11) ◽  
pp. 4234-4234
Author(s):  
Xiaoying Zhao ◽  
Lei Xu ◽  
Dong Wu ◽  
Rongzhen Xu
Keyword(s):  
Flow Cytometry ◽  
Cell Proliferation ◽  
Caspase 3 ◽  
K562 Cells ◽  
Human Leukemia ◽  
Dependent Manner ◽  
Flow Cytometry Assay ◽  
Transcript Levels ◽  
Induced Apoptosis ◽  
Dose Dependent

Abstract Purpose: To investigate apoptosis-inducing effects of Berbamine on human leukemia cells and to explore the underlying mechanism. Materials and methods: Berbamine was dissolved in 0.9% sodium chloride to an initial concentration of 1mg/ml and subsequently diluted to desired concentrations with cell culture medium. MTT was used to examine the effect of Berbamine on cell proliferation of K562 cells. Characteristic cellular morphological changes were used as indicators of apoptosis in K562 cells while the rate of apoptosis was measured by flow cytometry assay. Expression levels of apoptosis related genes bcl-2 and bax were determined by RT-PCR and the levels of bcr/abl were evaluated by nested-PCR. Levels of Caspase 3 were measured by flow cytometry assay. Results: Berbamine inhibited the cell proliferation significantly and in a dose-dependent manner in tested K562 cells. Its IC50 value was 5.23ug/ml. As determined by morphological observations and flow cytometry assay, Berbamine was able to induce apoptosis of K562 cells within 6 hours. The apoptosis rate of K562 was also dose-dependent. Steady-state transcript levels of bcr/abl decreased dramatically (half-quantity ratio from 1.284 to 0.506 within 72 hours following 8mg/ml Berbamine treatment. On the other hand, the protein levels of Caspase 3 surged from 18.36% to 38.25% (p<0.001) within 24 hours after treatment of 12mg/ml Berbamine. During the same period, no changes of bcl-2 or bax transcript levels were detected in the cells that were treated with 8mg/ml Berbamine. Conclusions: Our results suggest that Berbamine is a potent inhibitor of cell proliferation and a strong inducer of apoptosis in human K562 cells. The Berbamine-induced apoptosis pathway involves down regulation of bcr/abl and up regulation of Caspase 3 expressions. Neither bcl-2 nor bax plays substantial roles in Berbamine-induced K562 cell apoptosis.

scholarly journals Optimization and Limitations of Use of Cryopreserved Peripheral Blood Mononuclear Cells for Functional and Phenotypic T-Cell Characterization

2009 ◽  
Vol 16 (8) ◽  
pp. 1176-1186 ◽  
Author(s):  
Adriana Weinberg ◽  
Lin-Ye Song ◽  
Cynthia Wilkening ◽  
Anne Sevin ◽  
Bruce Blais ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Cell Viability ◽  
Lymphocyte Proliferation ◽  
Mononuclear Cells ◽  
Pokeweed Mitogen ◽  
Cell Recovery ◽  
Flow Cytometry Assay ◽  
Peripheral Blood Mononuclear ◽  
Affect Cell Viability ◽  
Blood Mononuclear Cells

ABSTRACT The goals of this study were to optimize processing methods of cryopreserved peripheral blood mononuclear cells (PBMC) for immunological assays, identify acceptance parameters for the use of cryopreserved PBMC for functional and phenotypic assays, and to define limitations of the information obtainable with cryopreserved PBMC. Blood samples from 104 volunteers (49 human immunodeficiency virus-infected and 55 uninfected) were used to assess lymphocyte proliferation in response to tetanus, candida, and pokeweed-mitogen stimulation and to enumerate CD4+ and CD8+ T cells and T-cell subpopulations by flow cytometry. We determined that slowly diluting the thawed PBMC significantly improved viable cell recovery, whereas the use of benzonase improved cell recovery only sometimes. Cell storage in liquid nitrogen for up to 15 months did not affect cell viability, recovery, or the results of lymphocyte proliferation assays (LPA) and flow cytometry assays. Storage at −70°C for ≤3 weeks versus storage in liquid nitrogen before shipment on dry ice did not affect cell viability, recovery, or flow cytometric results. Storage at −70°C was associated with slightly higher LPA results with pokeweed-mitogen but not with microbial antigens. Cell viability of 75% was the acceptance parameter for LPA. No other acceptance parameters were found for LPA or flow cytometry assay results for cryopreserved PBMC. Under optimized conditions, LPA and flow cytometry assay results for cryopreserved and fresh PBMC were highly correlated, with the exception of phenotypic assays that used CD45RO or CD62L markers, which seemed labile to freezing and thawing.

Nitroprusside induces cardiomyocyte death: interaction with hydrogen peroxide

2000 ◽  
Vol 279 (6) ◽  
pp. H3089-H3100 ◽  
Author(s):  
Simon W. Rabkin ◽  
Jennifer Y Kong
Keyword(s):  
Cell Death ◽  
Cell Viability ◽  
Mtt Assay ◽  
Cell Structure ◽  
No Production ◽  
Facs Analysis ◽  
Acridine Orange Staining ◽  
The Media ◽  
Bleb Formation ◽  
Induced Apoptosis

We examined the hypothesis that sodium nitroprusside (SNP) produces cell death in cardiomyocytes through generation of H2O2. Embryonic chick cardiomyocytes in culture were treated with SNP, and cell viability was assessed by trypan blue, MTT assay, and fluorescent activated cell sorting (FACS) analysis. SNP for 24 h induced a significant ( P < 0.001) dose-dependent loss of cell viability. On MTT assay, the half-maximal effective concentration was 0.53 mM (confidence interval 0.45–0.59 mM). SNP-treated cardiomyocytes displayed characteristic microscopic features of apoptosis: reduced cell size, nuclear disintegration, and membrane bleb formation. FACS analysis demonstrated SNP-induced apoptosis as well as cell changes consistent with necrosis. The proportion of cells with nuclear changes of apoptosis, identified by propidium iodide (PI) staining of permeabilized cells, increased significantly ( P < 0.05) after 0.5 mM SNP for 24 h. The proportion of apoptotic cells, characterized by dual staining of intact cardiomyocytes with fluorescein diacetate and PI, was significantly ( P < 0.05) increased after treatment with 0.5 mM SNP for 24 h. SNP metabolism and NO production was suggested by the significant ( P < 0.05) increase in nitrite generation in the media with 0.5 mM SNP compared with control. SNP-mediated H2O2 production was implicated in the mechanism of SNP-induced cell death. First, SNP produced a significant ( P< 0.05) increase in H2O2 detected in the media after 6 or 24 h of SNP treatment. Second, catalase completely blocked the reduction of cell viability induced by 0.1 mM SNP and significantly ( P < 0.05) blunted the effect of 0.5 mM SNP. In contrast, the iron chelator deferoxamine did not alter SNP-induced loss of cell viability. FACS analysis showed that the combination of low concentrations of H2O2(10−8 M) that did not alter cell viability augmented SNP-induced apoptosis. In contrast, the amount of necrotic cell death was unchanged by the combination of H2O2 and SNP. H2O2 plus SNP produced a dramatic alteration in cell structure with greater membrane bleb formation, shrunken cells, and more intense cytosolic acridine orange staining and nuclear fragmentation than either agent alone. These data indicate the vulnerability of cardiomyocytes to SNP and suggest the involvement of H2O2 in the pathogenesis of SNP-induced cardiomyocyte cell death. Establishing apoptosis as a component of the type of cell death induced by SNP permitted the recognition that SNP-induced apoptosis was increased by chronic treatment with low (subtoxic) concentrations of H2O2.

scholarly journals The oncolytic efficacy and safety of avian reovirus and its dynamic distribution in infected mice

2019 ◽  
Vol 244 (12) ◽  
pp. 983-991
Author(s):  
Ruimin Cai ◽  
Guangyuan Meng ◽  
Yi Li ◽  
Wenyang Wang ◽  
Youxiang Diao ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Liver Cancer ◽  
Mtt Assay ◽  
Avian Reovirus ◽  
Dapi Staining ◽  
Flow Cytometry Assay ◽  
Dynamic Distribution ◽  
Qrt Pcr ◽  
Ldh Assay ◽  
Oncolytic Activity

Primary liver cancer is a major public health challenge that ranks as the third most common cause of cancer worldwide despite therapeutic improvement. Reovirus has been emerging as a potential anti-cancer agent and is undergoing multiple clinical trials, and it is reported that reovirus can preferentially cause the cell death of a variety of cancers in a manner of apoptosis. As few studies have reported the efficacy of oncolytic activity and safety profile of avian reovirus, in our study, LDH assay, MTT assay, DAPI staining, and flow cytometry assay were performed to demonstrate the oncolytic effects of avian reovirus against the HepG2 cells, and quantitative real-time PCR (qRT-PCR) and animal experiments were conducted to investigate the dynamic distribution of avian reovirus in infected mice and then illustrate the safety and tissue tropism of avian reovirus. LDH assay, DAPI staining, and flow cytometry assay confirmed the efficacy of the oncotherapeutic effects of avian reovirus, and MTT assay has indicated that avian reovirus suppressed the proliferation of HepG2 cells and decreased their viability significantly. qRT-PCR revealed the dynamic distribution of avian reovirus in infected mice that avian reovirus might replicate better and have more powerful oncolytic activity in liver, kidney, and spleen tissues. Furthermore, histopathological examination clearly supported that avian reovirus appeared non-pathogenic to the normal host, so our study may provide the new insights and rationale for the new strategy of removing liver cancer. Impact statement We demonstrated the efficacy of oncolytic activity of avian reovirus (ARV) by LDH assay, MTT assay, DAPI staining, and flow cytometry assay, and also investigated the dynamic distribution of ARV in infected mice and then illustrated the safety and tissue tropism of ARV by quantitative real-time PCR (qRT-PCR) and animal experiments. Collectively, our study may provide the new insights and rationale for the new strategy of removing liver cancer.

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