Showing 12 results for Apoptosis
Malek Soleimani Mehranjani, Hamid Reza Momeni, Mohammad Hosein Abnosi, Parva Nasimi,
Volume 12, Issue 3 (10-2009)
Abstract
Abstract Background: Spinal cord slices culturing from adult mammals could be considered as a suitable in-vitro model for evaluating cellular viability, spinal cord injury and cell death mechanisms. In present study, determining of cell death in motor neurons of cultured spinal cord slices in adult mouse was done. Materials and Methods: In a experimental- laboratory study, thoracic regions of spinal cords from 4 Balb/c mice were cut into 400-µm slices using tissue chopper and incubated in a Co2 incubator at 37˚C for different periods of time. Freshly prepared slices (0h) and cultured slices were fixed and sectioned using cryostat. To study morphological and biochemical features of cell death, fluorescent staining, TUNEL method and agarose gel electrophoresis were used. Results: In freshly prepared slices of motor neurons showed no apoptotic changes. While, 6, 12 and 24h after culturing, this neurons displayed morphological features of apoptosis including cell shrinkage as well as nuclear and chromatin condensation. Also, 6 and 12h after culturing were TUNEL positive. In addition, extracted DNA from cultured slices for 24h were indicated the nucleosomal DNA fragmentation on agarose gel electrophoresis. Conclusion: Results were showed the occurrence of apoptosis in motor neurons of cultured adult mouse spinal cord slices.
Mohammad Amin Moosavi, Soroush Moasses Ghafary, Masood Asadi, Iraj Asvadi Kermani ,
Volume 14, Issue 4 (9-2011)
Abstract
Background: Leukemia is a malignant and progressive disease. Over-expression of inhibitors of apoptosis proteins (IAPs), such as survivin and its anti-apoptotic variants, including sur-ΔEx3, is the main cause of resistance to apoptotic effects of chemotherapy drugs. In the present study, the effects of CBX on apoptosis and expression level of survivin and sur-ΔEx3 and K562 cells (experimental model of chronic myeloid leukemia) were investigated.
Materials and Methods: In this experimental study, human K562 cells were cultured and exposed to CBX. Trypan blue exclusion test was used to evaluate growth inhibitory and viability effects of the drug. Fluorescent microscopy (acridine orange/ ethidium bromide double staining) and DNA electrophoresis were applied to the study of apoptosis. The expression level of survivin and sur-ΔEx3 was studied by semiquantative RT- PCR.
Results: The results showed that after the 48 h treatment of K562 cells with 150 µM CBX, significant growth inhibitory and apoptotic effects (up to 50%) were induced. In addition, after 2-4 h of treatment with CBX (150 µM), down-regulation of survivin and sur-∆Ex3 were observed. However, the expression level of survivin and sur-ΔEx3 increased to the level of control cells with longer treatment times (6-12 h).
Conclusion: Noticing the apoptotic and down-regulatory effects of CBX on survivin and sur-∆Ex3 expression, this drug can be used as a potential candidate for further studies on CML treatment, especially for inhibition of drug resistance in leukemia cells.
Mohammad Amin Moosavi , Soroush Moasses Ghafary, Masood Asadi, Iraj Asvadi Kermani ,
Volume 14, Issue 5 (11-2011)
Abstract
Background: To date, several drugs have been proposed for the treatment of acute promyelocytic leukemia (APL) however, none of them has resulted in complete remission. Therefore, many efforts are in progress to find new drugs with the capability of inducing apoptosis. Recently, anti-carcinogenic effects have been reported for a drug named carbenoxolone (CBX) on several cell lines. In the present study, the effects of CBX on NB4 cell line, as an experimental model of APL, were examined.
Materials and Methods: In this trial, NB4 cell line was cultured and treated with different concentrations of CBX (50-250µM) in various time intervals (12-48 hours). Trypan blue exclusion test was used to evaluate growth inhibitory and viability effects of the drug on NB4 cell line. Fluorescent microscopy (acridine orange/ethidium bromide double-staining) and agarose gel electrophoresis DNA were used to study apoptosis.
Results: CBX induced growth inhibition of NB4 cells so that growth inhibition rates of NB4 cells, after the 48 hour of treatment with 50, 100, 150, 200, and 250 µM CBX were 32.65, 47.52, 60.73, 68.91, and 74.33%, respectively. Furthermore, the results of DNA fragmentation and fluorescent microscopy assays indicated that apoptosis is a major mode of cell death after treatment of NB4 cells with above concentrations of CBX.
Conclusion: Noticing the growth inhibitory and apoptotic effects of CBX on human promyelocytic leukemia NB4 cells, it can be considered as a potential candidate for further studies on APL treatment.
Mohammad Amin Moosavi, Negin Seyed Gogani , Iraj Asvadi Kermani , Masood Asadi,
Volume 14, Issue 6 (1-2012)
Abstract
Background: Nucleostemin plays a critical role in controlling proliferation and self-renewal of stem cells and cancer cells. Thus, inhibition of nucleostemin expression could be a potent therapeutic approach in cancer treatment. In the present study, the effects of nucleostemin gene silencing in K562 cell line were studied.
Materials and Methods: In this experimental study, after transfecting NS-specific siRNA into K562 cells, changes in nucleostemin gene expression pattern were determined by semi-quantitative reverse transcription polymerase chain reaction (RT-PCR). Trypan blue exclusion test, MTT assay, and fluorescent microscopy were used to evaluate the growth inhibition and apoptosis of K562 cells, respectively. Flow-cytometery was utilized for evaluating the effects of nucleostemin gene silencing on cell cycle.
Results: The results showed the high expression of nucleostemin gene in K562 cells. NS-siRNA transfection into K562 cells at 200 nM inhibited the nucleostemin mRNA level up to 55% after 48 hours when compared to corresponding control cells. Forty eight hours after transfection, the cell growth decreased up to 33.7%. In addition, the silencing of nucleostemin induced G1 cell cycle arrest. Furthermore, fluorescent microscopy assays indicated that apoptosis occurred 48 hours after silencing nucleostemin gene expression.
Conclusion: Noticing the potent growth inhibitory and apoptotic effects of nucleostemin siRNA in human myeloid leukemia K562 cells, silencing this gene can be a potential target for inhibiting K562 cells as the stem cell model of chronic myeloid leukemia.
Davood Bashash, Seyed H. Ghaffari, Maryam Kazerani, Kebria Hezaveh, Kamran Alimoghaddam, Ardeshir Ghavamzadeh,
Volume 15, Issue 9 (2-2013)
Abstract
Background: Since nearly 90% of patients with acute promyelocytic leukemia (APL) have high telomerase activity and significant shortened telomere length, these patients have, therefore, been suggested to be good candidates for the therapeutic intervention with telomerase inhibitors. This study was done to investigate the effects of BIBR1532, a non-nucleoside inhibitor of telomerase, on APL cells. Materials and Methods: In this experimental study, for investigating the effect of BIBR1532, NB4 leukemic cells were cultured in the presence of various concentrations of BIBR1532. Succeeding apoptosis assay, Caspase-3 activity assay, and quantitative real-time PCR were applied to examine the effect of this drug on apoptosis percenage, enzymatic activity of Caspase-3, and quantitative expression of genes mRNA involved in apoptosis. Results: The results showed that BIBR1532 induced apoptosis in NB4 cells in a dose-dependent maner. Moreover, real time PCR results showed that BIBR1532 led to a significant decrease in mRNA of Bcl-2 gene and signficant increases in transcription of Bax, PUMA, and Caspase-3. Conclusion: Since treatment with BIBR1532 could exert rapid apoptotic cell death in NB4 cells andactivate cellular apoptosis route, anti-telomerase-based therapy can regarded as a suitable strategy for APL treatment. Patients with progressive shortening of telomere length and high levels of telomerase activity are suitable candidates for treatment with telomerase inhibitors.
Mina Mahdavi Rad, Nowruz Najafzadeh, Ali Niapour, Alireza Jafar,
Volume 17, Issue 6 (9-2014)
Abstract
Background: Melanoma is a malignant tumor of melanocytes, early-stage melanomas can be treated effectively with surgery alone, but more advanced cancers often incurable. The incidence of melanoma malignancy in most countries has risen faster than any other cancer types. It was the first time which we evaluated the cytotoxic effects of zno and Ag/zno nano-composites (NP) on melanoma cell line, A375, viability.
Materials and Methods: In this experimental study, A375 cell line was grown in RPMI-1640 supplemented with 10% FBS, penicillin/streptomycin (100 U/ml, 100 µg/ml) at 37◦C in 5% CO2, then the effects of different concentrations of zno and Ag/zno nano-composites on melanoma cell were evaluated by MTT, clonogenic survival assays, and acridine orange/ ethidium bromide staining.
Results: Herein, we demonstrated that Zno and Ag/zno nano-composites showed similar effects on cytotoxicity of melanoma cancer cells. In a dose dependent manner, a significant cytotoxicity was observed with increasing of zno and Ag/zno. The inhibitory concentration 50% (IC50) values of the nano-composites for A375 cell line after 24 hrs were 7.24±1.55 and 15.93±1.73 µg/ml for zno and Ag/zno, respectively.
Conclusion: The results showed that zno and Ag/zno has ability to induce cytotoxicity in the human melanoma cancer cell line in lower micromolar concentrations. In conclusion, these findings may introduce a new view on the mode of action and possible application of new nano-composites in the cancer chemotherapy.
Malek Soleimani Mehranjani, Majid Mahdiyeh, Atena Sadat Azimi,
Volume 18, Issue 7 (10-2015)
Abstract
Background: Alpha-tocopherol, as a strong antioxidant, plays an important role in testraining free radicals. The aim of this study was to investigate the effect of Alpha-tocopherol on cell proliferation and restraining apoptosis in rat bone marrow mesenchymal stem cells.
Materials and Methods: In this research study, the rat bone marrow mesenchymal stem cells were extracted under sterile conditions using flashing-out method. At the end of the third passage, cells were divided into groups of control and Alpha-tocopherol with doses of 15 and 25 µM and were treated in the osteogenic media cell medium containing 10% fetal bovine serum, 10 mM β-glycerol phosphate, 10 nM dexamethasone and 50 µg/ml ascorbic 3-phosphate] for a period of 21 days. Then, cell proliferation, DNA damage, expression of Bcl-2 and Bax genes and the morphologic changes of the cells were investigated during the procedure of osteogenesis. Data were analyzed using one-way ANOVA and means difference was considered significant at p<0.05.
Results: Cell proliferation, the size of nuclei diameter and expression of anti-apoptotic Bcl-2 gene showed a significant increase in mesenchymel stem cells treated with Alpha-tocopherol (p<0.05) in a dose dependent manner compared to the control cells. Also, cytoplasm extension was seen in the cells treated with Alpha-tocopherol, compared to the control group. Since Alpha-tocopherol causes a significant decrease in DNA damage and the expression of apoptotic Bax gene, compared to the control group, therefore it can suppress apoptosis in bone marrow mesenchymal stem cells, in a dose dependent manner .
Parvin Farzanegi, Masoumeh Habibian, Hadi Alinejad,
Volume 19, Issue 3 (6-2016)
Abstract
Background: Chronic kidney disease as an important risk factor is associated with some disorders which are key causes of death and disability in older people. Therefore, the aim of this study was to assess the combined effect of regular aerobic exercise with garlic extract on renal apoptosis regulatory factors in aged rats with chronic kidney disease.
Materials and Methods: In this experimental research, 42 aged male Wistar rats(48-52 weeks) were selected and randomly divided into 6 groups: control, doxorubicin, doxorubicin-salin, doxorubicin- garlic, doxorubicin - exercise, doxorubicin –garlic-exercise(combined). Chronic kidney disease was induced by a single subcutaneous injection 8.5 mg/kg of doxorubicin. Swimming training was programmed 3 days /week, 30 min/day for 8 weeks. Both the doxorubicin garlic and combined groups with garlic extract were administered by garlic gavage at a dose of 2.5 g/kg. The renal Bax and Bcl-2 levels were evaluated by ELIZA method. A one-way analysis of variance was used to data analysis (p<0.05).
Results: The results showed that induced chronic kidney disease was associated with a significant increase on Bax and a decrease on Bcl-2 in aged rats. Also, 8 weeks swimming training, garlic supplementation and the combined intervention significantly reversed these changes. Furthermore, no significant difference have been observed in the effect of these interventions on Bax and Bcl-2 in aged rats with chronic kidney disease.
Conclusion: It seems that the use of non-pharmacological treatment methods such as exercise training, garlic extract supplement, and combination of the both interventions may be effective in reducing apoptosis resulted from chronic kidney disease in aged rats.
Nasrin Kzemipour, Seyed Mehdi Shariatzadeh, Saeed Nazifi,
Volume 19, Issue 9 (12-2016)
Abstract
Abstract
Background: Silver nanoparticles are capable of inducing toxicity in living organisms. Silver nanoparticles can induce some effect in the liver. Thus silver nanoparticles, due to their wide spread effects, can also affect on hepatic, hematological, and oxidative stress factors. Ginger because of its powerful antioxidantal compounds can influence the toxicity effects of silver nanoparticles in different parts of the body. The aim of this study was to investigate the protective effect of hydroalchoholic extract of ginger on cytotoxic silver nanoparticles on enzymes, hematological parameters, blood oxidative stress markers, and hepatic apoptosis in Balb-c mice.
Materials and Methods: In this study, 48 rats of Balb-c race Syrians were selected and devided into 4 groups, each consisting of 12. They were treated for a period of 35 days; the first group (control) received distilled water, the second group received nano silver, the third group received ginger extract, and the fourth group received both nanosilver and ginger extract at the same time. Bleeding was done to measure hematological factors, liver enzymes, and oxidative stress; then liver tissue was removed for evaluation of apoptosis. Data were compared using SPSS software and one-way ANOVA.
Results: Enzymes AST , ALT , ALP, GGT and LDH as liver factors showed significant differences in the groups of the study. Hematological factors including of WBC , RBC , Hb , HCT , MCV, MCH , Plt , Lymphocyte and Monocyte showed significant differences in all the groups.
Of oxidative stress factors , only GPX showed significant difference between groups, while no significant difference was observed in other oxidative stress parameters in the blood. Changes in apoptosis showed significant differences in all groups of the study.
Conclusion: Based on the findings the study ,silver nanoparticles with their side effects in different parts of the body can induce changes in various factors and enzymes. Ginger can compensate ,and modify to some extent these side effects. Such effectiveness of ginger can probably be due to its special ingredients.
Maryam Salem, Abolfazl Bayrami, Tooba Mirzapour, Mohsen Sagha,
Volume 21, Issue 1 (4-2018)
Abstract
Abstract
Background: According to application of Retinoic acid in differentiation of the stem cells to different cells and its role in apoptotic of cancer cells, the selection of appropriate dose for differentiation of stem cells is important. Thus in this study the effects of Retinoic acid in different concentrations on viability stem cells to select the appropriate dose for differentiation was investigated.
Materials and Methods: In this study, bone marrow mesenchymal stem cells were affected by different concentrations of Retinoic acid. Survival of cells was investigated after 3, 10 and 15 days of culture by MTT assay. DAPI staining was used to evaluate the number of apoplectic nuclei in treated cells after 10 and 15 days.
Results: After three days of culture, the results showed that a large number of cells are destroyed at concentrations of 10-4, 10-3 and 10-2M of Retinoic acid, while in 10-5 and 10-6 M of Retinoic acid, it is not observed many apoptosis. Amount of 10-5M Retinoic acid after 10 days showed significant apoptosis, while the concentration of 10-6 M Retinoic acid after 15 days showed significant apoptosis compared to the control group (p<0.05).
Conclusion: It looks that 10-6 M Retinoic acid is an appropriate concentration for differentiation of mesenchymal stem cells.
Nima Sanadgol , Mohammad Sharifzadeh , Parisa Maleki ,
Volume 22, Issue 3 (8-2019)
Abstract
Background and Aim: Regarding the importance of new treatments to control and treat multiple sclerosis (MS), in this study we investigated the role of Benzoaric acid (BA) on the neuro-inflammation and apoptosis processes in the cuprizone (cup)-induced animal model of MS.
Materials and Methods: In this experimental study, 35 males C57BL/6 mice were divided into five groups. The study groups were included, control: received six weeks of normal powdered food beside intraperitoneal (i.p.) injection of BA solvent (100 µL per day PBS) for the last two weeks, cup: received six weeks of powdered food contains 0.2% cup beside i.p. injection of BA solvent for the last two weeks and cup-treatment: received six weeks of powdered food contains 0.2% cup beside i.p. injection of 20, 40 and 80 mg/kg BA for the last two weeks. Eventually, the medial corpus callosum area of the animal’s brain was evaluated via western blot and Real-Time PCR methods.
Ethical Considerations: Ethical points were observed according to the declaration of Helsinki and relevant code of ethics, regarding minimizing harms during animal experimentation (UOZ-GR-9517-13).
Findings: Molecular studies have shown that BA-80 decreased mRNA (p <0.01) and protein expression of NF-KB and consequently increased I-KB/NF-KB ratio (p <0.05) and decreased inflammation in compare to cup group. Moreover, BA-80 decreased caspase-9 mRNA (p<0.01) and caspase-8 mRNA (p <0.05) and subsequently increased caspase-8/caspase-9 ratio (p<0.01) and decreased apoptosis in compare to cup group.
Conclusion: The dose of 80 mg/ml BA via decreasing cup-induced neuro-inflammation and neuro-apoptosis has protective effects in this model.
Sahar Dehghani, Leila Rouhi, Noosha Ziya Jahromi, Reza Dehghani, Khalil Khashei Varnamkhasti,
Volume 24, Issue 2 (5-2021)
Abstract
Background and Aim: Proliferate potential differentiate into different cell lineages and high self-renewal of Mesenchymal Stem Cells (MSCs); thus, they are ideal tools for regenerative medicine. However, a leading problem is an oxidative stress in the target tissue and the apoptosis of transplanted stem cells before tissue repair. The pretreatment of stem cells with antioxidants may make them resistant to oxidative stress. Ginger is the main medicinal plant with antioxidant properties. This study explored the antioxidant effects of ginger extract on bioavailability and oxidative stress-induced apoptosis in human adipose tissue-derived mesenchymal stem cells and rat bone marrow examined.
Methods & Materials: In this study, human adipose tissue-derived mesenchymal stem cells and rat bone marrow were cultured in a DMEM medium with 20% FBS. The explored cells were incubated for 4 and 6 hours for pretreatment with different concentrations of ginger extract (50, 100, 200, & 400 mg/mL); then, they were treated with 200 μM H2O2 for 2 hours. Bioavailability was analyzed by ELISA reader using an MTS kit and apoptosis was analyzed by flow cytometry using an Annexin V-FITC/PI kit into the manufacturer’s protocol at both times. The obtained data were analyzed by Analysis of Variance (ANOVA) using SPSS.
Ethical Considerations: This study was approved by the Ethics Research Committee of Shahrekord Branch, Islamic Azad University (Code: IR.IAU.SHK.REC.1397.028).
Results: The MTS results indicated a dose- and time-dependent manner increase in the bioavailability of human adipose tissue-derived mesenchymal treated stem cells. Ginger extract treatment also dose- and time-dependently decreased the rate of apoptosis in rat bone marrow mesenchymal stem cells.
Conclusion: Ginger extract, by reducing the oxidative stress in mesenchymal stem cells, elevates their lifespan in the target tissue, and increases the efficiency of these cells in tissue regeneration.
