No abstract

The prefrontal cortex is the largest lobe of the brain and is consequently involved in stroke. There is no comprehensive practical pharmacological strategy for ameliorating prefrontal cortex injury induced by cerebral ischemia. Therefore, we studied the neuroprotective properties of verapamil (Ver) on mitochondrial dysfunction and morphological features of apoptosis in transient global ischemia/reperfusion (I/R). Ninety-six Wistar rats were allocated into four groups: control, I/R, I/R+Ver (10 mg/kg twice 1 hour prior to ischemia and 1 hour after reperfusion phase), and I/R+NaCl (vehicle). Animals were sacrificed, and mitochondrial dysfunction parameters (i.e., mitochondrial swelling, mitochondrial membrane potential, ATP concentration, ROS production, and cytochrome c release), antioxidant defense (i.e., superoxide dismutase, malondialdehyde, glutathione peroxidase, catalase, and caspase-3 activation), and morphological features of apoptosis were determined. The results showed that mitochondrial damage, impairment of antioxidant defense system, and apoptosis were significantly more prevalent in the I/R group in comparison with the other groups. Ver decreased mitochondrial damage by reducing oxidative stress, augmented the activity of antioxidant enzymes in the brain, and decreased apoptosis in the I/R neurons. The current study confirmed the role of oxidative stress and mitochondrial dysfunction in I/R progression and indicated the possible antioxidative mechanism of the neuroprotective activities of Ver.

Background: Methamphetamine (MA), is an extremely addictive stimulant that adversely affects the central nervous system. Accumulating evidence indicates that molecular mechanisms such as oxidative stress, apoptosis, and autophagy are involved in the toxicity of MA. Considering experimental animal studies exhibiting MA-induced neurotoxicity, the relevance of these findings needs to be evidently elucidated in human MA users. It is generally assumed that multiple chemical substances released in the brain following MA-induced metabolic activation are primary factors underlying damage of neural cells. Hence, this study aimed to investigate the role of autophagy and apoptosis as well as oxidative stress in the brain of postmortem MA-induced toxicity. Materials and Methods: In this study, we determine the gene expression of autophagy and apoptosis, including BECN1, MAP1ALC3, CASP8, TP53, and BAX genes in ten healthy controls and ten chronic users of MA postmortem dorsolateral prefrontal cortex (DLPFC) by real-time polymerase chain reaction. Also, we applied immunohistochemistry in formalin-fixed and paraffin-embedded human brain samples to analyze brain-derived neurotrophic factor (BDNF). Also, spectrophotometry was performed to measure glutathione (GSH) content. Results: The expression level of apoptotic and autophagic genes (BECN1, MAP1ALC3, CASP8, TP53, and BAX) were significantly elevated, while GSH content and BDNF showed substantial reductions in DLPFC of chronic MA users. Discussion: Our data showed that MA addiction provokes transduction pathways, namely apoptosis and autophagy, along with oxidative mechanisms in DLPFC. Also, MA induces multiple functional and structural perturbations in the brain, determining its toxicity and possibly contributing to neurotoxicity. [GMJ.2021;10:e2016]

Background: Gliomas possess low immunogenicity, which is an inevitable hinder in front of cancer immunotherapy. Different interferons (IFNs) may proceed apoptosis instead in p53-dependent or independent pathways. P53 induces the anti-inflammatory programmed cell death in cancer cells; on the other hand, IFN gamma (IFNγ) is a modulatory/pro-inflammatory cytokine. There are contradictory reports of whether this cytokine can possess an anti- or pro-cancerous impact on tumors. Hence, we aimed to investigate the possible cooperative apoptotic effect of the P53 and IFNγ over expressions on the U87 glioblastoma cell line. Materials and Methods: The P53 expressing vector was amplified by Escherichia coli BL21. This vector was confirmed by the aid of sequencing. At the next step, U87 cells were transfected using lipofectamine. Cells were treated with P53 vector and/or IFNγ. The type of cellular death investigated by flow cytometry and the expression level of cleaved caspase-3 protein was also precisely demonstrated by western blotting. Results: Sequencing results revealed that inserted P53 was identical with human P53. Western blot results revealed that both IFNγ and P53 overexpression could up-regulate cleaved caspase-3 protein expression in this cell line. Interestingly, flow cytometry data determined that concurrent treatment with P53 exogenous overexpression and IFNγ induces about 70% apoptosis in U87; more than the sum of cell death occurs after IFNγ or P53 overexpression alone (~18%+21%=39%). Conclusion: The present study results showed that p53-overexpression and IFNγ could ultimately induce up-regulation of the caspase-3 and ultimately significant apoptosis increasing in the U87 cell line. Although IFNγ is believed to be a pro-inflammatory cytokine and P53 is an anti-inflammatory agent, our results demonstrated that they could act synergistically to induce apoptosis in U87 cells. [GMJ.2021;10:e2270]

Background: Osteosarcoma is known as the malignant tumors of bone. Cyanidin 3-O-glucoside (C3G) has a powerful ability to induce apoptotic cell death in the different cancer cells however the mechanisms of action for C3G have not been clarified yet. Objective: In this study, we have investigated the apoptotic effects of C3G on three different osteosarcoma cell lines including Saso-2, MG-63, G-292 (clone A141B1). Methods: The 24 hours IC50 of C3G for Saso-2, G-292, and MG-63 cells was evaluated by MTT assay. Apoptosis induction in these cell lines after treatment with C3G approved by the way of Annexin V/PI flow cytometry. Changes at the mRNA expression level of PPARγ, P21, Bax, and Bcl-xl genes have investigated by real-time PCR technique, as well as P21 expression was further confirmed by western blotting. Results: The MTT assay results have demonstrated that the 24 hours IC50 of C3G for Saso-2 and G-292 cells was 110μg/ml while it was about 140μg/ml for the MG-63 cells. The results of real-time PCR have clearly shown that treatment of the cells with 24 hours IC50 of C3G case to upregulation of PPARγ, P21, and Bax genes. Also, western blot analyzing confirmed that P21 protein overexpressed endogenously after treatment of cells with C3G which it was more upregulated in MG-63 cells compared to the other cell lines. Conclusion: The final outcomes introduce the C3G as a novel anti-osteosarcoma agent with the ability to induce apoptosis in different osteosarcoma cells via upregulation of the PPARγ and the P21.

Background: Atorvastatin is a member of statins, which has shown positive vascular effects, anti-oxidant, antiplatelet, and anti-apoptotic properties. Objective: In this study, we hypothesized that atorvastatin could prevent the neurons lost in the hippocampal dentate gyrus region after transient global ischemia/reperfusion (I/R) through its anti-oxidant and anti-apoptotic activities. Method: Twenty-four male Wistar rats 12-13 weeks old and weighing 250–300 g, were divided randomly into four groups: control, I/R, vehicle (I/R treated with NaCl) and experiment (I/R treated with atorvastatin, 10 mg/kg) and rats were sacrificed 96 hours after I/R. Quantitative expression of genes (caspase 8, p53, bax, bcl2, cytochrome c) was studied. The MDA level, SOD, CAT, and GPx activities were measured with biochemical tests. To detect apoptotic cells, TUNEL and Nissl staining were performed. Mitochondria were prepared from the hippocampus rats, used to the quantification of mitochondrial ROS, ATP level, GSH content, membrane potential, cytochrome c release, and determination of mitochondrial swelling. Results: Atorvastatin attenuated the overexpression of bax, cytochrome C, p53, and caspase8 mRNAs and induced expression of bcl-2 mRNA (P<0.001). Atorvastatin treatment increased anti-oxidant enzyme levels (P<0.01). Treatment with atorvastatin reduced the number of TUNEL-positive cells. It could decrease the cytochrome c release (P<0.01), inhibit the decrease of MMP (P<0.001) and increased the ATP level (P<0.001) in mitochondrial hippocampal in compared with I/R group. Conclusion: Atorvastatin treatment in I/R rats decreases oxidative stress, production of ROS, apoptosis rate in neuronal cells, and improves the mitochondrial function. Hence, atorvastatin have a proper neuronal protective effect against the I/R injury in the brain.

During the last months of the coronavirus pandemic, with all those public restrictions and health interventions, the transmission of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) appears now to have been raised in some countries around the world. Iran was one of those first countries facing the second wave of coronavirus, due to the lack of appropriate public restrictions because of economic problems the country is facing. The clinical and demographic characteristics of severe cases and non-severe cases of Coronavirus Disease (COVID-19) in 192 patients in Tehran, Iran, between June 16 and July 11, 2020, were investigated. The patients were divided into severe cases (n = 82) and non-severe cases (n = 110). Demographic and clinical characteristics were compared between the two study clusters. The mean age was 54.6 ± 17.2 years, and the most common presenting symptom was persistent cough (81.8%) and fever (79.7%). The logistic regression model revealed that age, BMI, and affected family members were statistically associated with severity. Patients with complicated conditions of disorders faced more hospitalization days and medical care than the average statistical data. As the coronavirus spike in the case and death reports from June 2020, we observed the rise in the incidence of severe cases, where 42.7% (82/192) of cases have resulted in severe conditions. Our findings also suggested that the effect of IFB (Betamethasone) was more valid than the other alternative drugs such as LPV/r and IVIg.

Background: Resveratrol is a phenolic natural product which is found in red grapes also in Japanese knotweed root (Polygonum cuspidatum). Naringenin is one of the flavonoid compounds found in landing grape and other citrus fruits. Both agents exert antioxidant and anti-inflammatory properties. Objective: In this study, we investigated the effect of Resveratrol and Naringenin in an in vitro model of retinoblastoma of the eye. Method: XTT and trypan blue assays used to evaluate the anti-proliferative/cytotixic effect of resveratrol and naringenin in Y79 cells. By the aid of AnnexinV/PI flow cytometry the kind of cell death investigated. To asses important gene expression level at mRNA level involve in apoptosis, Real-time PCR utilized. Results: Naringenin and resveratrol significantly decreased proliferation and stimulated cell death (mostly apoptosis) in Y79 cells at 50 and 100 (μg/ml) after 24 and 48 hours. More cytotoxic effect observed after 48 hours. Furthermore expression level of Bax and Bcl2 mRNAs altered significantly in all samples treated with 50 (μg/ml) of naringenin, resveratrol, or simultaneously with both. P21 mRNAs expression altered in all mentioned samples except those treated with 50 (μg/ml) of resveratrol. Results: Naringenin and resveratrol significantly decreased proliferation and stimulated cell death (mostly apoptosis) in Y79 cells at 50 and 100 (μg/ml) after 24 and 48 hours. More cytotoxic effect observed after 48 hours. Furthermore expression level of Bax and Bcl2 mRNAs altered significantly in all samples treated with 50 (μg/ml) of naringenin, resveratrol, or simultaneously with both. P21 mRNAs expression altered in all mentioned samples except those treated with 50 (μg/ml) of resveratrol. Conclusion: Based on the results, it can be concluded that resveratrol and naringenin can decrease cell viability in retinoblastoma cells in an in vitro dose/time-dependent manner. Albeit more studies needed to shed the light on the mechanism of action, our data reveals a potential synergistic cytotoxic effect of naringenin and resveratrol on Y79 cells in 48 hours.