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1.
Iran J Basic Med Sci ; 27(7): 914-922, 2024.
Article En | MEDLINE | ID: mdl-38800026

Objectives: Adult neurogenesis, the process of generating new neurons, continues throughout life. Unfortunately, this process is insufficient in pathological conditions and needs to be promoted. Crocin, the active component of saffron, affects neurogenesis in vivo and in vitro. We aimed to investigate the enhancing effects of crocin on the neurogenesis of adipose-derived mesenchymal stem cells in the presence of retinoic acid, as well as the molecular pathways involved. Materials and Methods: Differentiation capacities and stemness potential of harvested ADSCs were evaluated by differentiating into osteocytes and adipocytes, and expression of mesenchymal CD markers by flow cytometry. The optimum dose of crocin was assessed with an MTT assay. Crocin, retinoic acid, CREB/BDNF, and Notch inhibitors and their combination were added to the culture medium. Jag1, Hes1, Notch, and BDNF gene expression were analyzed by RT-PCR on days 7, 14, and 21, while CREB, DCX, SOX2, and NeuN expression were analyzed by immunofluorescence. Results: Expression of mesenchymal CD markers as well as adipogenic and osteogenic differentiation confirmed the origin and properties of ADSCs. The optimal dose of crocin was 1 mM. Crocin significantly (P<0.05) increased, while inhibitors (DATP&Naphthol) significantly (P<0.05) decreased Jag1, Hes1, Notch, and BDNF expression. Immunofluorescent assessments showed that expression of DCX, BDNF, NeuN, and Sox2 proteins increased significantly (P<0.05) after crocin administration and decreased significantly (P<0.05) after inhibitor administration. Conclusion: Crocin can be used as an enhancer for neural differentiation of MSCs in vitro in the presence of retinoic acid. The mechanism is proposed through Notch and CREB/BDNF signaling pathways.

2.
Curr Radiopharm ; 17(1): 55-67, 2024.
Article En | MEDLINE | ID: mdl-38817005

BACKGROUND: Exposure to physical contamination during chemotherapy, including non-ionizing electromagnetic fields, raises concerns about the widespread sources of exposure to this type of radiation. Glioblastoma multiforme (GBM) is an aggressive central nervous system tumor that is hard to treat due to resistance to drugs such as temozolomide (TMZ). OBJECTIVE: Electromagnetic fields (EMF) and haloperidol (HLP) may have anticancer effects. In this study, we investigated the effects of TMZ, HLP, and EMF on GBM cell lines and analyzed the association between non-ionizing radiation and the risk of change in drug performance. METHODS: Cell viability and reactive oxygen species (ROS) generation were measured by MTT and NBT assay, respectively. Then, the expression levels of breast cancer-resistant protein (BCRP), Bax, Bcl2, Nestin, vascular endothelial growth factor (VEGF) genes, and P53, Bax, and Bcl2 Proteins were evaluated by real-time PCR and western blot. RESULTS: Co-treatment of GBM cells by HLP and TMZ enhanced apoptosis in T-98G and A172 cells by increasing the expression of P53 and Bax and decreasing Bcl-2. Interestingly, exposure of GBM cells to EMF decreased apoptosis in the TMZ+HLP group. CONCLUSION: In conclusion, EMF reduced the synergistic effect of TMZ and HLP. This hypothesis that patients who are treated for brain tumors and suffer from depression should not be exposed to EMF is proposed in the present study. There appears to be an urgent need to reconsider exposure limits for low-frequency magnetic fields, based on experimental and epidemiological research, the relationship between exposure to non-ionizing radiation and adverse human health effects.


ATP Binding Cassette Transporter, Subfamily G, Member 2 , Apoptosis , Cell Survival , Electromagnetic Fields , Haloperidol , Neoplasm Proteins , Nestin , Temozolomide , Vascular Endothelial Growth Factor A , Humans , Apoptosis/drug effects , Apoptosis/radiation effects , Nestin/metabolism , Temozolomide/pharmacology , Haloperidol/pharmacology , Vascular Endothelial Growth Factor A/metabolism , ATP Binding Cassette Transporter, Subfamily G, Member 2/metabolism , Cell Line, Tumor , Cell Survival/drug effects , Cell Survival/radiation effects , Neoplasm Proteins/metabolism , Neoplasm Proteins/biosynthesis , Glioblastoma/radiotherapy , Glioblastoma/metabolism , Reactive Oxygen Species/metabolism , Brain Neoplasms/radiotherapy , Brain Neoplasms/metabolism , Antineoplastic Agents, Alkylating/pharmacology , Glioma/radiotherapy , Glioma/metabolism , Glioma/pathology
3.
Diagn Pathol ; 19(1): 35, 2024 Feb 16.
Article En | MEDLINE | ID: mdl-38365810

BACKGROUND: Breast cancer is one of the most common diseases worldwide that affects women of reproductive age. miR-221 and miR-222 are two highly homogeneous microRNAs that play pivotal roles in many cellular processes and regulate the Wnt/ß-catenin signaling pathway. Curcumin (CUR), a yellow polyphenolic compound, targets numerous signaling pathways relevant to cancer therapy. The main aim of this study was to compare the ability of chitosan curcumin nanoparticle (CC-CUR) formulation with the curcumin in modulating miR-221 and miR-222 expression through Wnt/ß-catenin signaling pathway in MCF-7, MDA-MB-231 and SK-BR-3 breast cancer cell lines. METHOD: Chitosan-cyclodextrin-tripolyphosphate containing curcumin nanoparticles (CC-CUR) were prepared. Cytotoxicity of the CUR and CC-CUR was evaluated. Experimental groups including CC-CUR, CUR and negative control were designed. The expression of miR-221 and miR-222 and Wnt/ß-catenin pathway genes was measured. RESULTS: The level of miR-221 and miR-222 and ß-catenin genes decreased in MCF-7 and MDA-MB-231 cells and WIF1 gene increased in all cells in CC-CUR group. However, the results in SK-BR-3 cell line were unexpected; since miRs and WIF1 gene expressions were increased following CC-CUR administration and ß-catenin decreased by administration of CUR. CONCLUSION: Although the composite form of curcumin decreased the expression of miR-221 and miR-222 in MCF-7 and MDA cells, with significant decreasing of ß-catenin and increasing of WIF1 gene in almost all three cell lines, we can conclude than this formulation exerts its effect mainly through the Wnt/ß-catenin pathway. These preliminary findings may pave the way for the use of curcumin nanoparticles in the treatment of some known cancers.


Breast Neoplasms , Chitosan , Curcumin , MicroRNAs , Female , Humans , beta Catenin/genetics , beta Catenin/metabolism , Breast Neoplasms/drug therapy , Breast Neoplasms/genetics , Breast Neoplasms/metabolism , Cell Line, Tumor , Cell Proliferation , Chitosan/pharmacology , Curcumin/pharmacology , MCF-7 Cells , MicroRNAs/metabolism , Wnt Signaling Pathway/genetics , Nanoparticles
4.
J Ovarian Res ; 16(1): 238, 2023 Dec 15.
Article En | MEDLINE | ID: mdl-38102694

BACKGROUND: Polycystic ovary syndrome (PCOS) is a widespread heterogeneous disease that is in association with genetic, epigenetic, endocrine and environmental factors. Adipose-derived mesenchymal stem cell (ASC) and ASC-conditioned medium (ASC-CM) have shown promising abilities in tissue regeneration. In the present study, we aimed to investigate the effects of ASC and ASC-CM on epigenetic regulators, steroidal function and folliculogenesis in the letrozole-induced PCOS rats. RESULTS: Based on the measurement of the oral glucose tolerance test and physical parameters including body weight, estrus cycle pattern as well as ovary dimensions, PCOS-induced rats in sham and control (CTRL) groups showed signs of reproductive dysfunctions such as lack of regular estrus cyclicity, metabolic disorders such as increased ovary dimension, body weight and blood glucose level alteration which were improved especially by ASC-CM administration.


Mesenchymal Stem Cells , Polycystic Ovary Syndrome , Female , Humans , Rats , Animals , Culture Media, Conditioned/adverse effects , Body Weight , Mesenchymal Stem Cells/metabolism , Epigenesis, Genetic
5.
Basic Clin Neurosci ; 14(2): 263-272, 2023.
Article En | MEDLINE | ID: mdl-38107534

Introduction: Glioblastoma multiforme (GBM) is an aggressive case of primary brain cancer which remains among the most fatal tumors worldwide. Although, some in vitro and in vivo models have been developed for a better understanding of GBM behavior; a natural model of GBM would improve the efficiency of experimental models of human GBM tumors. We aimed the present study to examine the survival and durability of U87 cells in the brain of wild-type rats. Methods: U87 cells were intracranially implanted in twenty-one wild-type rats. Tumor size and morphology as well as infiltration of immune cells were investigated at three-time points by H&E and immunohistochemistry (IHC). Results: The results demonstrated that the inoculation of GBM cells led to the infiltration of host defense system cells which caused immunological regression of the tumor mass after six weeks. While the tumors successfully developed without any sign of host defense invasion in the second week of GBM inoculation. Also, a decrease in tumor size and infiltration of immune system cells were observed in the fourth week. Conclusion: These data remarkably suggest that time plays a crucial role in activating the immune system against human GBM tumors in rats; it shows that the regression of tumor mass depends on a time slope. Highlights: A noticeable proliferation of tumor cells was observed in the rat's brain by the second week.The distant metastatic masses of cancer infiltrated into the adjacent normal tissue by the second week.Tumor mass underwent a noticeable diminution in the size by the fourth week.Cancer cells completely regressed by the sixth week due to immunological reactions.In tumor rejection, the effective mechanism depends on immune system activity and the slope of time. Plain Language Summary: One of the most malignant tumors is the brain tumor in the world. Unfortunately, no effective treatment has yet been found for it. Of course, researchers need efficient animal models to find the appropriate treatment. The xenograft model is one of the tumor models in the laboratory. However, the main challenge is the interaction of the animal's immune system with induced-cancer cells so that the immune system finally rejects the tumor. In this study, we investigated how long the immune system needs to reject induced tumors in the xenograft model completely. For this purpose, we studied the animals in three periods (second week, fourth week, and sixth week). We concluded that the immune system does not recognize the induced cancer cells until the second week of the experiment. It results in the growth of cancer cells and the formation of tumors in the animal brain. However, the immune system begins to recognize the tumor mass after the fourth week which leads to a reduction in metastasis and tumor size. Eventually, the immune system completely rejects the formed tumor in the sixth week.

6.
Biochem Biophys Res Commun ; 679: 179-190, 2023 10 30.
Article En | MEDLINE | ID: mdl-37703761

Since the developmental stage of oocyte is a challenging issue in the success of vitrification, this study investigated the effects of vitrification, before and after in vitro maturation, on the survival and maturation rates, developmental competence and the expression levels of genes involved in apoptosis, oxidative stress and epigenetic modifications. Mouse germinal vesicle (GV) oocytes were divided into four groups: fresh in vitro matured oocytes without vitrification (fIVM), in vitro matured oocytes after vitrification (vIVM), in vitro matured oocytes before vitrification (IVMv). In addition, in vivo matured oocytes (MII) were used as control. After oocytes collection, maturation and survival rates as well as the intracellular reactive oxygen species (ROS) level were evaluated. Also, the expression level of various genes was analyzed by qRT-PCR. In addition, following artificial activation (parthenogenesis), the developmental competence of oocytes to the blastocyst stage was evaluated. A significant decrease in maturation rate and survival of vIVM oocytes was observed compared to fIVM and IVMv oocytes. Intracellular ROS levels were significantly increased in both vitrified groups compared to the fIVM group, and no significant difference between vitrified groups. Pro-apoptotic genes; BAX and Bcl2 as well as genes related to oxidative stress response Hsp1a, Hsp1b and SOD1were significantly increased in the vIVM group compared to the IVMv group. Interestingly, epigenetic regulators genes DNMT1, DNMT3a and DNMT3b were highly expressed in IVMv oocytes along with a decrease in the artificial activation rate compared to the vIVM oocytes. Our results indicated that despite observing more negative effects of vitrification before IVM on the survival rate and maturation as well as apoptosis status, less epigenetic changes in vIVM oocytes can make this process a better option in the treatment of infertility than IVM of oocytes followed by vitrification, a hypothesis that needs to be investigation in human oocytes.


In Vitro Oocyte Maturation Techniques , Vitrification , Humans , Animals , Mice , In Vitro Oocyte Maturation Techniques/methods , Reactive Oxygen Species , Oocytes , Cryopreservation/methods , Oxidative Stress
7.
Gen Comp Endocrinol ; 344: 114370, 2023 12 01.
Article En | MEDLINE | ID: mdl-37611672

AIMS: this study aimed to examine the protective role of omega-3 and insulin on reproductive system of the male mouse model of type II diabetes mellitus (T2DM), especially DNA integrity and chromatin quality. MAIN METHODS: adult age-matched mice were divided into intact, sham, or T2DM groups (n = 7) which received a high-fat diet/low-dose streptozotocin. T2DM-induced animals underwent no treatment as diabetic control (T2DM), received omega-3 (T2DM + Omg3), received insulin (T2DM + Ins) and their combination (T2DM + Omg3 + Ins) for 35 days. After which, testicular and sperm parameters and testosterone levels were evaluated. KEY FINDINGS: our findings revealed that the various examined parameters were comparable between the intact and sham groups, while most testicular and sperm parameters were affected by T2DM. Treatment of T2DM-induced animals with omega-3, alone and in combination with insulin, significantly improved sperm motility, normal morphology, sperm chromatin quality, DNA integrity, Leydig cell number and non-significantly testosterone levels. SIGNIFICANCE: T2DM interferes with spermatogenesis and steroidogenesis as well as sperm quality and DNA integrity, which can be partially ameliorated by long-term administration of omega-3 in combination with or without insulin. Although our findings should be confirmed in clinical studies, since previous clinical trials have found omega-3 consumption to be beneficial in humans, its use seems to be safe and effective.


Diabetes Mellitus, Type 2 , Insulin , Humans , Adult , Male , Mice , Animals , Insulin/pharmacology , Diabetes Mellitus, Type 2/drug therapy , Mice, Inbred C57BL , Sperm Motility , Semen , Testis , Testosterone/pharmacology , Chromatin , DNA
8.
BMC Complement Med Ther ; 23(1): 45, 2023 Feb 14.
Article En | MEDLINE | ID: mdl-36788561

BACKGROUND: Early-life exposure to exogenous estrogens such as phytoestrogens (plant-derived estrogens) could affect later health through epigenetic modifications. Foeniculum vulgare (fennel) and Linum usitatissimum (flax) are two common medicinal plants with high phytoestrogen content. Considering the developmental epigenetic programming effect of phytoestrogens, the main goal of the present study was to evaluate the perinatal exposure with life-long exposure to hydroalcoholic extracts of both plants on offspring's ovarian epigenetic changes and estrogen receptors (ESRs) expression level as signaling cascades triggers of phytoestrogens. METHODS: Pregnant mice were randomly divided into control (CTL) that received no treatment and extract-treated groups that received 500 mg/kg/day of fennel (FV) and flaxseed (FX) alone or in combination (FV + FX) during gestation and lactation. At weaning, female offspring exposed to extracts prenatally remained on the maternal-doses diets until puberty. Then, the ovaries were collected for morphometric studies and quantitative real-time PCR analysis. RESULTS: A reduction in mRNA transcripts of the epigenetic modifying enzymes DNMTs and HDACs as well as estrogen receptors was observed in the FV and FX groups compared to the CTL group. Interestingly, an increase in ESRα/ESRß ratio along with HDAC2 overexpression was observed in the FV + FX group. CONCLUSION: Our findings clearly show a positive relationship between pre and postnatal exposure to fennel and flaxseed extracts, ovarian epigenetic changes, and estrogen receptors expression, which may affect the estrogen signaling pathway. However, due to the high phytoestrogen contents of these extracts, the use of these plants in humans requires more detailed investigations.


Flax , Foeniculum , Plant Extracts , Prenatal Exposure Delayed Effects , Animals , Female , Mice , Pregnancy , Epigenesis, Genetic , Estrogens , Flax/adverse effects , Foeniculum/adverse effects , Ovary , Phytoestrogens/adverse effects , Plant Extracts/adverse effects , Receptors, Estrogen/metabolism
9.
Psychopharmacology (Berl) ; 240(1): 77-86, 2023 Jan.
Article En | MEDLINE | ID: mdl-36385208

RATIONALE: Opioid use disorders are commonly treated by long-acting agonist opioids including methadone and buprenorphine which could affect various aspects of male reproduction especially spermatogenesis. OBJECTIVES: We aimed to determine whether detoxification with methadone or buprenorphine was associated with reproductive disorders in male mice. METHODS: We orally induced morphine dependence in NMRI male mice, and then performed detoxification programs using either methadone or buprenorphine. Testis architecture and sperm parameters including sperm nuclear DNA integrity, mitochondrial activity, oxidative stress in seminal plasma, and routine sperm parameters were assessed to find the involved mechanisms. RESULTS: The number of Leydig cells and the thickness of germinal epithelium reduced following morphine use and increased differently after detoxification with methadone or buprenorphine. Morphine dependence and detoxification with methadone and buprenorphine had different effects on sperm parameters. Morphine altered chromatin integrity, mitochondrial activity, and oxidative stress in sperm. Detoxification with methadone improved mitochondrial activity but worsened chromatin integrity, whereas detoxification with buprenorphine improved neither chromatin integrity nor mitochondrial activity. Seminal plasma oxidative stress was higher in the treated groups compared to control groups but was comparable among treatment groups. Our study revealed that long-term morphine use followed by detoxification with methadone or buprenorphine impairs testis structure and sperm parameters. Detoxification from morphine use with methadone and buprenorphine led to different preclinical outcomes in semen quality parameters, including chromatin integrity. Therefore, clinical detoxification protocols should be performed more cautiously, considering the desire of the individuals to reproduce.


Buprenorphine , Morphine Dependence , Opioid-Related Disorders , Male , Mice , Animals , Methadone/therapeutic use , Buprenorphine/pharmacology , Buprenorphine/therapeutic use , Opiate Substitution Treatment , Morphine Dependence/drug therapy , Semen Analysis , Semen , Analgesics, Opioid/pharmacology , Analgesics, Opioid/therapeutic use , Morphine , Opioid-Related Disorders/drug therapy , Reproduction , Chromatin
10.
Biochem Biophys Res Commun ; 637: 259-266, 2022 12 31.
Article En | MEDLINE | ID: mdl-36410275

Alteration of the proliferation rate and differentiation of mesenchymal stem cells (MSCs) into various lineages, including neural cells, by light emitting diodes (LEDs) irradiation has received considerable attention in recent years. Human umbilical cord matrix-derived mesenchymal cells (hUCMs) are an accessible source of adult stem cells with appropriate characteristics that make them ideal tools for stem cell researches, cell therapy procedures and regenerative medicine. The aim of the present study was to investigate the effects of green LED irradiation, retinoic acid (RA) and their combination on the differentiation of hUCMs into neural lineage as well as the mechanisms involved. Exposure of hUCMs to green LED (530 nm, 1.59 J/cm2) with or without retinoic acid (RA) treatment, significantly increased the expression of specific genes including nestin, ß-tubulin III, MAP2 and Olig2. In addition, immunohistochemical analysis confirmed expression of specific neural-related proteins including MAP2, GFAP and Olig2 in irradiated cells. ROS generation significantly increased following green light irradiation which in turn has activated the MAPK signaling pathway, resulting in the differentiation of hUCMs into neurons and glial cells, confirmed by western blot analysis of MAPK-related pathway. Taken together, our results suggest that the green LED irradiation, alone and in combination with RA, via ERK 1/2, JNK and p38 phosphorylation improves differentiation of hUCMs into neural lineage. Other mechanisms and inducers to enhance differentiation phenomena in vitro and in vivo should be investigated to determine the most appropriate strategy for therapeutic purposes.


Mesenchymal Stem Cells , Umbilical Cord , Adult , Humans , Tretinoin/pharmacology , Cell Differentiation , Phosphorylation
11.
Andrologia ; 54(11): e14593, 2022 Dec.
Article En | MEDLINE | ID: mdl-36123787

Green LED and three-dimensional (3D) scaffolds have recently received extensive attentions due to their impact on cell proliferation and differentiation. Melatonin, a circadian rhythm-regulating hormone, is involved in some physiological phenomena including testosterone biosynthesis. Lower testosterone biosynthesis results in some disorders such as puberty retarding, andropause, and muscle weakness. Therefore, our aim was to investigate the proliferation of Leydig cells and their testosterone-related Gene expression and secretion under the influence of 3D scaffold, green light and melatonin. The experimental groups of TM3 cells embedded in the 3D scaffold, were exposed to green light, melatonin, both and all three factors. Expression of cell cycle genes including PCNA, CYCLIND1, CDC2 and CDKN1B, and testosterone related genes; GATA4 and RORα were also examined. 3D scaffold enhanced Leydig cells proliferation, and testosterone-related genes expression. While melatonin decreased cell proliferation and testosterone-related genes expression. Green light did not significantly change the results but slightly decreased cell proliferation and testosterone synthesis. The combination of green light with melatonin significantly reduced the proliferation rate of TM3 cells and the expression of steroidogenic genes, while the combination of green light with scaffold improved the results. In general, the use of scaffolding enhances proliferation and testosterone-related genes expression of TM3 Leydig cells. Also, application of green light and scaffolding reduces the deleterious effects of melatonin on these cells.


Leydig Cells , Melatonin , Male , Humans , Melatonin/pharmacology , Sexual Maturation , Testosterone/metabolism , Cell Proliferation
12.
Int J Reprod Biomed ; 19(10): 889-898, 2021 Oct.
Article En | MEDLINE | ID: mdl-34805729

BACKGROUND: Vitrification as the most efficient method of cryopreservation, enables successful storage of oocytes for couples who undergo specific procedures including surgery and chemotherapy. However, the efficacy of in vitro maturation (IVM) methods with vitrified germinal vesicle (GV) oocytes could be improved. OBJECTIVE: As melatonin and follicular fluid (FF) might enhance IVM conditions, we used these supplements to assess the maturation rate of vitrified GV oocytes and their artificial fertilization rate. MATERIALS AND METHODS: Four hundred mouse GV oocytes were harvested, vitrified, and assigned into control (C-Vit-GV) and treatment groups of melatonin (M-Vit-GV), human follicular fluid (HFF-Vit-GV), and a combination (M + HFF-Vit-GV). A non-vitrified group of GV oocytes (non-Vit-GV) and a group of in vivo matured metaphase II (Vivo-MII) oocytes served as control groups to evaluate the vitrification and IVM conditions, respectively. Maturation of GV oocytes to MII and further development to two-cell-stage embryos were determined in the different groups. RESULTS: Development to two-cell embryos was comparable between the Vivo-MII and non-Vit-GV groups. IVM and in vitro fertilization (IVF) results in the non-Vit-GV group were also comparable with the C-Vit-GV oocytes. In addition, the IVM and IVF outcomes were similar across the different treatment groups including the M-Vit-GV, HFF-Vit-GV, M + HFF-Vit-GV, and C-Vit-GV oocytes. CONCLUSION: Employing an appropriate technique of vitrification followed by suitable IVM conditions can lead to reasonable IVF outcomes which may not benefit from extra supplementations. However, whether utilizing other supplementation formulas could improve the outcome requires further investigation.

13.
Toxicol Ind Health ; 37(11): 674-684, 2021 Nov.
Article En | MEDLINE | ID: mdl-34644184

Mancozeb (MZB) is a worldwide fungicide for the management of fungal diseases in agriculture and industrial contexts. Human exposure occurs by consuming contaminated plants, drinking water, and occupational exposure. There are reports on MZB's reprotoxicity such as testicular structure damage, sperm abnormalities, and decrease in sperm parameters (number, viability, and motility), but its molecular mechanism on apoptosis in testis remains limited. To investigate the molecular mechanisms involved in male reprotoxicity induced by MZB, we used primary cultures of mouse Sertoli-germ cells. Cells were exposed to MZB (1.5, 2.5, and 3.5 µM) for 3 h to evaluate viability by 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) assay, reactive oxygen species (ROS) generation, and oxidative stress parameters (lipid peroxidation). Cell death and mitogen-activated protein kinase (MAPK) signaling were measured in these cells using flow cytometry and western blotting. In addition, some groups were exposed to N-acetylcysteine (NAC, 5 mM) in the form of co-treatment with MZB. Mancozeb reduced viability and increased the level of intracellular ROS, p38 and c-Jun N-terminal kinases (JNK) MAPK proteins phosphorylation, and apoptotic cell death, which could be blocked by NAC as an inhibitor of oxidative stress. The present study indicated for the first time the toxic manifestations of MZB on the Sertoli-germ cell co-culture. Redox imbalance and p38 and JNK signaling pathway activation might play critical roles in MZB-induced apoptosis in the male reproductive system.


Apoptosis/drug effects , Maneb/toxicity , Mitogen-Activated Protein Kinases/pharmacology , Sertoli Cells/drug effects , Zineb/toxicity , Animals , Germ Cells/drug effects , Male , Mice , Oxidative Stress/drug effects
14.
Asian Pac J Cancer Prev ; 22(8): 2471-2478, 2021 Aug 01.
Article En | MEDLINE | ID: mdl-34452560

OBJECTIVE: Angiogenesis plays a dominant role in many pathophysiologic disorders, including cancer. Tranilast, which is an anti-fibrotic drug, is also suggested as an anti-angiogenesis agent. As Teucrium polium (TP) is known as an herbal medicine with antitumor properties, this study aimed to investigate the effects of TP and Tranilast on human umbilical vein endothelial cells (HUVECs), in vitro model of angiogenesis, as well as rat's aortic ring ex vivo model. METHODS: In this study, The HUVECs were treated with various doses of TP and Tranilast each one alone or in combination together. Cell survival test, aortic ring ex-vivo assay, and evaluating mRNA expressions of VEGFA and TGF-ß ligands and receptors were performed. RESULTS: The survival rate of HUVECs has significantly (p <0.05) reduced by TP and Tranilast. The combination of both TP and Tranilast significantly reduced cell viability as compared to the administration of TP or Tranilast alone. As well, the treatment of HUVECs with TP and/or Tranilast significantly (p <0.05) decreased TGF-ß1, TGF-ß 2, TGF-ßRI, and TGF-ßRII mRNA expression levels, but not the expression of TGF-ß3 and TGF-ßRIII in the TP-treated cells. Image analysis showed that TP and/or Tranilast inhibited vascular growth in the aortic ring assay. CONCLUSION: Our results strongly support the anti-angiogenic effects of the TP and Tranilast combination on both in vitro and ex vivo models of angiogenesis. However, further investigations in in vivo models and human studies are needed before human use.


Angiogenesis Inhibitors/pharmacology , Fibrin/chemistry , Human Umbilical Vein Endothelial Cells/drug effects , Neovascularization, Pathologic/drug therapy , Plant Extracts/pharmacology , Teucrium/chemistry , ortho-Aminobenzoates/pharmacology , Angiogenesis Inhibitors/chemistry , Animals , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Drug Synergism , Drug Therapy, Combination , Human Umbilical Vein Endothelial Cells/pathology , Humans , Male , Neovascularization, Pathologic/metabolism , Neovascularization, Pathologic/pathology , Rats , Rats, Sprague-Dawley , Receptor, Transforming Growth Factor-beta Type I/genetics , Receptor, Transforming Growth Factor-beta Type I/metabolism , Transforming Growth Factor beta1/genetics , Transforming Growth Factor beta1/metabolism , ortho-Aminobenzoates/chemistry
15.
Sci Rep ; 11(1): 12311, 2021 06 10.
Article En | MEDLINE | ID: mdl-34112894

It is well established that maternal lifestyle during pregnancy and lactation affects the intrauterine programming of F1 offspring. However, despite the co-use of alcohol and nicotine is a common habit, the effects of exposure to both substances on the reproductive system of F1 male offspring and the underlying mechanisms of developmental programming have not been investigated. The present study aimed to examine pre- and postnatal concurrent exposure to these substances on genetic and epigenetic alterations of sperm cells as well as testis properties of F1 offspring compared with exposure to each substance alone. Pregnant dams in the F0 generation randomly received normal saline, nicotine, ethanol, and combinations throughout full gestation and lactation periods. Sperm cells and testes of F1 male offspring were collected at postnatal day 90 for further experiments. High levels of sperm DNA fragmentation were observed in all exposed offspring. Regarding epigenetic alterations, there was a significant increase in the relative transcript abundance of histone deacetylase 1 and 2 in all exposed sperm cells. Moreover, despite a decrease in the expression level of DNA methyltransferase (DNMT) 3A, no marked differences were found in the expression levels of DNMT1 and 3B in any of the exposed sperm cells compared to non-exposed ones. Interestingly, combined exposure had less prominent effects relative to exposure to each substance alone. The changes in the testicular and sperm parameters were compatible with genetic and epigenetic alterations. However, MDA level as an oxidative stress indicator increased in all exposed pups, which may be responsible for such outputs. In conclusion, maternal co-exposure to these substances exhibited epigenotoxicity effects on germline cells of F1 male offspring, although these effects were less marked relative to exposure to each substance alone. These counteracting effects may be explained by cross-tolerance and probably less impairment of the antioxidant defense system.


Prenatal Exposure Delayed Effects/genetics , Sperm Motility/drug effects , Spermatozoa/drug effects , Testis/physiopathology , Animals , DNA Copy Number Variations/genetics , DNA Fragmentation , DNA Methyltransferase 3A , Epigenesis, Genetic/drug effects , Ethanol/adverse effects , Female , Humans , Lactation/drug effects , Male , Maternal Exposure/adverse effects , Mice , Nicotine/adverse effects , Pregnancy , Prenatal Exposure Delayed Effects/physiopathology , Reproduction/genetics , Sperm Count , Spermatozoa/pathology
16.
Front Pharmacol ; 12: 830331, 2021.
Article En | MEDLINE | ID: mdl-35126154

Crocin, the main biologically active carotenoid of saffron, generally is derived from the dried trifid stigma of Crocus sativus L. Many studies have demonstrated that crocin has several therapeutic effects on biological systems through its anti-oxidant and anti-inflammatory properties. The wide range of crocin activities is believed to be because of its ability to anchor to many proteins, triggering some cellular pathways responsible for cell proliferation and differentiation. It also has therapeutic potentials in arthritis, osteoarthritis, rheumatoid arthritis, and articular pain probably due to its anti-inflammatory properties. Anti-apoptotic effects, as well as osteoclast inhibition effects of crocin, have suggested it as a natural substance to treat osteoporosis and degenerative disease of bone and cartilage. Different mechanisms underlying crocin effects on bone and cartilage repair have been investigated, but remain to be fully elucidated. The present review aims to undertake current knowledge on the effects of crocin on bone and cartilage degenerative diseases with an emphasis on its proliferative and differentiative properties in mesenchymal stem cells.

17.
Drug Des Devel Ther ; 14: 3589-3599, 2020.
Article En | MEDLINE | ID: mdl-32943846

PURPOSE: Human butyrylcholinesterase (BChE) serves as a bio scavenger to counteract organophosphate poisoning. It is also a potential drug candidate in several therapeutic fields. Therefore, in the present study, we constructed a new dual-promoter plasmid consisting of Cytomegalovirus (CMV) and human elongation factor 1α (EF-1α) promoters and transfected that into HEK-293 cells using Lipofectamine to enhance the BChE secretion. METHODS: The new dual-promoter construction (pBudCE dual BChE) including two copies of the BChE gene was designed and transfected into cells by liposomal structures. The cloned plasmids were evaluated by enzyme digestion and gel electrophoresis analysis. Experimental groups were categorized into the cells transfected by pBudCE dual BChE (treatment), pCMV (positive control) vectors, and nontransfected cells (negative control). BChE gene expression was evaluated by qRT-PCR and the enzyme activity was assessed using modified Ellman's method. The freeze-thaw process was carried out for analyzing the stability of the pBudCE dual BChE vector. RESULTS: Validation examination of the cloned plasmids confirmed the successful cloning process. The gene expression level and Ellman's method value in pBudCE dual BChE was higher than the other groups. CMV promoter has also increased the enzyme activity, although the difference was not significant compared with the control group. Interestingly, freeze-thaw cycles followed by several passages did not affect the enzyme activity. CONCLUSION: The designed construction with CMV and EF-1α promoters could increase BChE gene expression and the activity of the BChE enzyme in HEK-293 cell line. Large-scale production of BChE enzyme can be achieved by using dual-promoter plasmid construction compared to a single-promoter vector to be used in clinical trials.


Butyrylcholinesterase/metabolism , Lipids/genetics , Butyrylcholinesterase/genetics , Cells, Cultured , Cytomegalovirus/genetics , Genetic Vectors/genetics , HEK293 Cells , Humans , Peptide Elongation Factor 1/genetics , Promoter Regions, Genetic/genetics
18.
Drug Des Devel Ther ; 14: 3087-3097, 2020.
Article En | MEDLINE | ID: mdl-32801647

PURPOSE: Genetic diseases can be the result of genetic dysfunctions that happen due to some inhibitory and/or environmental risk factors, which are mostly called mutations. One of the most promising treatments for these diseases is correcting the faulty gene. Gene delivery systems are an important issue in improving the gene therapy efficiency. Therefore, the main purpose of this study was modifying graphene oxide nanoparticles by spermine in order to optimize the gene delivery system. METHODS: Graphene oxide/APTES was modified by spermine (GOAS) and characterized by FT-IR, DLS, SEM and AFM techniques. Then pEGFP-p53 was loaded on GOAS, transfected into cells and evaluated by fluorescent microscopy and gene expression techniques. RESULTS: FT-IR data approved the GOAS sheet formation. Ninety percent of the particles were less than 56 nm based on DLS analysis. SEM analysis indicated that the sheets were dispersed with no aggregation. AFM results confirmed the dispersed structures with thickness of 1.25±0.87 nm. STA analysis showed that GOAS started to decompose from 400°C and was very unstable during the heating process. The first weight loss up to 200°C was due to the evaporation of absorbed water, the second one observed in the range of 200-550°C was assigned to the decomposition of labile oxygen- and nitrogen-containing functional groups, and the third one above 550°C was attributed to the removal of oxygen functionalities. In vitro release of DNA demonstrated the efficient activity of the new synthesized system. Ninety percent of the cells were transfected and showed the GFP under fluorescence microscopy, and TP53 gene was expressed 51-fold in BT-20 cells compared to ß-actin as the reference gene. Flow cytometry analysis confirmed the apoptosis of the cells rather than necrosis. CONCLUSION: It could be concluded that the new synthesized structure could transfer a high amount of the therapeutic agent into cells with best activity.


Breast Neoplasms/therapy , Gene Transfer Techniques , Genetic Therapy , Graphite/chemistry , Nanoparticles/chemistry , Propylamines/chemistry , Silanes/chemistry , Spermine/chemistry , Breast Neoplasms/genetics , Female , Green Fluorescent Proteins/genetics , Humans , Microscopy, Fluorescence , Spectroscopy, Fourier Transform Infrared , Transfection , Tumor Cells, Cultured , Tumor Suppressor Protein p53/genetics
19.
Biochem Biophys Res Commun ; 514(1): 239-245, 2019 06 18.
Article En | MEDLINE | ID: mdl-31029424

Recently, light emitting diode (LED) irradiation has been introduced as a new strategy to enhance proliferation and affect differentiation of stem cells. Human Wharton's jelly-derived mesenchymal (hWJM) cells have unique characteristics that make them an appropriate source of stem cells for use in basic and clinical applications. In this study, we aimed to evaluate the effect of polarized (PL) and non-polarized (NPL) red light irradiation on gametogenic differentiation of hWJM cells in the presence or absence of bone morphogenetic protein 4 (BMP4) and retinoic acid (RA). Exposure of hWJM cells to PL and NPL red LED (625 nm, 1.9 J/cm2) with or without BMP4+RA pre-treatment effectively differentiated them into germ lineage when the gene expression pattern (Fragilis, DAZL, VASA, SCP3 and Acrosin) and protein synthesis (anti-DAZL, anti-VASA, anti-SCP3 and anti-Acrosin antibodies) of the induced cells was evaluated. These data demonstrated that photobiomodulation may be applied for gametogenic differentiation in-vitro.


Mesenchymal Stem Cells/cytology , Wharton Jelly/cytology , Acrosin/genetics , Bone Morphogenetic Protein 4/pharmacology , Cell Cycle Proteins/genetics , Cell Differentiation , Cells, Cultured , DEAD-box RNA Helicases/genetics , DNA-Binding Proteins/genetics , Gene Expression Regulation/drug effects , Germ Cells/cytology , Germ Cells/physiology , Humans , Light , Mesenchymal Stem Cells/drug effects , Mesenchymal Stem Cells/physiology , RNA-Binding Proteins/genetics , RNA-Binding Proteins/metabolism , Tretinoin/pharmacology
20.
Adv Pharm Bull ; 9(1): 76-83, 2019 Feb.
Article En | MEDLINE | ID: mdl-31011561

Purpose: To investigate the protective effect of preconditioning with non-toxic dose of hydrogen peroxide (H2O2) as a possible cell signaling molecule, against cell death induced by toxic concentration of H2O2 or by serum deprivation in human Wharton's jelly-derived mesenchymal stem cells (HWJ-MSCs) and underlying mechanisms. Methods: HWJ-MSCs were isolated and identified using flow cytometry. After finding non-toxic concentration of H2O2, cells preconditioning was performed by H2O2 (20 µM) for 12 h and cell tolerance against serum deprivation or toxic levels of H2O2 was assayed by MTT test. Effect of preconditioning on mRNA and protein expression of Akt-1, Bcl-2 and Bax were examined using reverse transcription polymerase chain reaction (RT-PCR) and western blotting respectively. Role of hypoxia-inducible factor (HIF)-1α was explored in presence HIF-1α inhibitor. Results: Preconditioning with 20 µM H2O2 for 12 h was non-toxic and decreased cell death induced by oxidative stress and serum deprivation in MSC cultures. However, the increased tolerance reversed in the presence of inhibitor of HIF-1α. By regards to RT-PCR and western blotting data, although expression of Akt-1, Bcl-2 and Bax was not change considerably but phosphorylated Akt-1 (pAkt-1) was up regulated after treatment with 20 µM H2O2 compared to control group. Moreover after exposure to 100 µM H2O2, western blotting analysis showed that cell pretreatment with 20 µM H2O2, decremented Bax/Bcl2 ratio and up-regulated HIF-1α and pAkt-1 compared to the control group. Conclusion: Increased tolerance of H2O2-pretreated cells led to the suggestion that transplantation of H2O2 preconditioned MSCs may improve therapeutic potential of stem cells in cell therapy procedures.

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