Bax Gene Expression Research Articles

Effect of Dioscorea extract on Bax and Bcl-2 gene expression in MCF-7 and HFF cell lines

BackgroundIn cancer cells, the balance between proliferation and apoptosis is disturbed. There is a direct relationship between gene expression and the process of apoptosis. The two genes involved in apoptosis are Bax and Bcl-2, and it is now well established that some plant compounds can alter the expression of genes. The aim of this study is to determine the rate of change in the expression of these genes in the cell line MCF-7 treated with Dioscorea extract for 24, 48 and 72 h. For this purpose, the plant extract was prepared by Soxhlet method and diluted in different concentrations. MCF-7 and HFF cell lines were treated in three replicates with different concentrations of the extract at intervals of 24, 48, and 72 h. To evaluate the toxicity of the extract, the MTT assay was performed and the IC50 value was calculated. Both cell types were cultured at IC50 concentration with three treatments and three replicates. RNA extraction, cDNA synthesis and real-time PCR were then performed. Flow cytometry was performed to further confirm apoptosis.ResultsMTT results showed that 72 h treatment with Dioscorea extract in IC50 concentration had the greatest effect on the death of MCF-7 cancer cells, while the cells of the control cell line remained healthy. The results of the study of gene expression changes showed that when treated with the plant extract for 24 h, the increase in Bax gene expression and the decrease in Bcl-2 gene expression were not statistically significant. At 48-h treatment, the decrease in Bcl-2 expression was not statistically significant, whereas the increase in Bax expression, which was 2.1 times, was statistically significant. When treated with the plant extract for 72 h, Bax expression increased 2.72 times and Bcl-2 gene expression decreased 0.67 times. Flow cytometry showed that 72-h treatment with plant extract at a concentration of 438.35 µg/ml was the most effective treatment for MCF-7 cancer cell death.ConclusionsThe expression ratio of Bax gene to Bcl-2 is equal to 4.06, which indicates the induction of more apoptosis by treatment with plant extract.

Study the Anticancer Properties of Thymol-Loaded PEGylated Bovine Serum Albumin Nanoparticles Conjugated with Folic Acid.

Phenolic compounds such as Thymol have an effective role in suppressing cancer, however, their low solubility in aqueous solution has limited their use. This study aimed to prepare Thymol (TY)-loaded bovine serum albumin (BSA) nanoparticles surface-modified with polyethylene glycol (PEG) conjugated with folic acid (FA) and evaluate their inhibitory activity on cancer cells. The TY-BSA-PEG-FA was characterized using DLS, FESEM, and FTIR. The encapsulation efficiency (EE) was evaluated indirectly by using UV absorption. The antioxidant property of nanoparticles was evaluated by 2,2'-azinobis(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS), 2,2-diphenyl-1-picrylhydrazyl (DPPH), and ferric reducing ability of plasm (FRAP) methods. The effects of nanoparticles against cancer cells were investigated by MTT, AO/PI, flow cytometry, and real-time qPCR methods. The results showed the spherical morphology of TY-BSA-PEG-FA with an average size of 70.0 nm, a PDI of 0.32, a zeta potential of -11.3 mV, and an EE of 89.0±2.3 %. The cytotoxicity effects of nanoparticles against all cell lines were in a concentration-dependent manner. AGS gastric cancer cells were reported to be the most vulnerable to treatment, while pancreatic cancer cells (PANC-1) and normal skin cells (HFF) would be the most resistant. The SubG1 phase arrest of about 66 % occurred at 85 μg/mL. An increase in apoptotic cells in fluorescent staining, along with decreased expression of Bcl-2 and increased expression of the BAX gene demonstrated the induction of apoptosis in treated cells. The powerful inhibitory effect of nanoparticles in inhibiting ABTS free radicals (IC50 =82 μg/mL) and DPPH free radicals (IC50 =844 μg/mL) and the ability to reduce iron ions indicated the antioxidant effects of TY-BSA-PEG-FA. Based on these results, the synthesized nanoparticles may be suitable for further investigation in the treatment of cancer, notably gastric cancer.

Comparison of cytotoxic, apoptotic and oxidative properties of Akacid plus and chlorhexidine in corneal epithelial cell culture.

This study aims to compare the cytotoxic, apoptotic, and oxidative effects of a new cationic disinfectant, Akacid Plus, with chlorhexidine, on the human corneal epithelial cell line. Time-dependent cytotoxicity studies were performed with the Alamar Blue method. Apoptotic activity was investigated by flow cytometric methods. Reactive oxygen species levels were measured with the ROS cellular test kit. BAX, BCL2 and caspase 3, 9, 12 mRNA expressions were evaluated by PCR, as well as BAX and BCL2 protein expressions by Western-Blot. At the fifth minute of the treatment, the viability was 68.15% with Akacid Plus and 43.95% with chlorhexidine. At the 15th minute, no significant difference was observed with both solutions. In the apoptotic evaluation, Akacid Plus significantly increased the early and late apoptotic activity in the cell line (p < 0.0001), while a significant increase was observed in late apoptosis and necrosis levels with chlorhexidine (p < 0.001). Chlorhexidine also induced gene expression of BAX, BCL2, caspase 3, 9 and BAX proteins (p < 0.05), while reducing protein expression of BCL2 (p < 0.001). Akacid Plus induced the gene expressions of BCL2, CASP3 and caspase 9, reduced gene expressions of BAX and caspase 12 and protein expression of BCL2 (p < 0.05). No significant difference was observed in the ROS level with both solutions (p > 0.05). Due to the widespread use of cationic polymers in ophthalmology, this new molecule with high antimicrobial activity and relatively low cytotoxicity may be of interest for clinical use. Further investigations are necessary to fully understand the ophthalmologic potential of this solution.

Caffeine-folic acid-loaded-chitosan nanoparticles combined with methotrexate as a novel HepG2 immunotherapy targeting adenosine A2A receptor downstream cascade

BackgroundMethotrexate (MTX) is a common chemotherapeutic drug that inhibits DNA synthesis and induces apoptosis. Treatment with MTX increased CD73 expression, which leads to higher levels of extracellular adenosine. Adenosine levels are also high in the tumor microenvironment through Cancer cells metabolism. That promotes the survival of cancer cells and contributes to tumor immune evasion through the Adenosine 2a Receptor. A2A receptor antagonists are an emerging class of agents that treat cancers by enhancing immunotherapy, both as monotherapy and in combination with other therapeutic agents. Caffeine is an adenosine receptor antagonist. Herein, we demonstrate the ability of a novel well prepared and characterized nano formula CAF-FA-CS-NPs (D4) for A2aR blockade when combination with MTX to improve its antitumor efficacy by enhancing the immune system and eliminating immune suppression.MethodsCAF-FA-CS-NPs (D4) were prepared and characterized for particle size, loading efficiency, and release profile. Molecular docking was used to validate the binding affinity of caffeine and folic acid to A2A receptor. The effects of the nano formula were evaluated on human liver cancer cells (HepG2), breast cancer cells (MCF-7), and MDA-MB-231, as well as normal human cells (WI-38). Different combination ratios of MTX and D4 were studied to identify the optimal combination for further genetic studies.ResultsMolecular docking results validated that caffeine and folic acid have binding affinity to A2A receptor. The CS-NPs were successfully prepared using ionic gelation method, with caffeine and folic acid being loaded and conjugated to the nanoparticles through electrostatic interactions. The CAF loading capacity in D4 was 77.9 ± 4.37% with an encapsulation efficiency of 98.5 ± 0.37. The particle size was optimized through ratio variations. The resulting nanoparticles were fully characterized. The results showed that (D4) had antioxidant activity and cytotoxicity against different cancer cells. The combination of D4 with MTX (IC50 D4 + 0.5 IC50 MTX) resulted in the downregulation of Bcl-2, FOXP3, CD39, and CD73 gene expression levels and upregulation of Bax and A2AR gene expression levels in HepG2 cells.ConclusionsThis study suggests that CAF-FA-CS-NPs (D4) in combination with MTX may be a promising candidate for cancer immunotherapy, by inhibiting A2aR signaling and leading to improved immune activation and anti-tumor activity of MTX.

The protective impact of curcumin, vitamin D and E along with manganese oxide and Iron (III) oxide nanoparticles in rats with scrotal hyperthermia: Role of apoptotic genes, miRNA and circRNA

BackgroundInfertility is one of the major factors affecting most people around the world. Short-term exposure to high temperatures can cause hyperthermia, which is one of the causes of male infertility. The aim of this study was to investigate the protective effect of curcumin, vitamins D and E along with Iron (III) oxide nanoparticles (Fe2O3-NPs) and manganese oxide nanoparticles (MnO2-NPs) on semen parameters and its effect on miRNA21 and circRNA0001518 expression. Material and methodsIn this study, the lower part of the rat was exposed to 43 °C for 5 weeks every other day for 5 weeks. Then the animals were killed. Tissue samples were collected for sperm parameters analysis, and tissue samples were taken for evaluation of apoptosis levels in germ cells, and RNA extraction in order to examine the expression of Bax, Bcl-2, miRNA, and CircRNA genes. ResultsThe results of this study showed that administration of curcumin, vitamin D, and vitamin E with Fe2O3-NPs and MnO2-NPs can improve the parameters of semen, Bax gene expression, Bcl-2 as well as miRNA and CircRNA in rats with testicular hyperthermia. In addition, curcumin by reducing the toxicity of Fe2O3 nanoparticles was able to reduce its negative effects and also reduce apoptosis in germ cells. This decrease in apoptosis was attributed to decreased Bcl-2 gene expression and increased expression of Bax, miRNA-21, and circRNA0001518. ConclusionAll the results of this study confirmed that Fe2O3-NPs and Mno2-NPs containing antioxidants or vitamins are useful in improving fertility in rats due to scrotal hyperthermia. Although Fe2O3-NPs and Mno2-NPs containing both antioxidants and vitamins had a greater effect on improving fertility and reducing the toxic effects of nanoparticles.

Helicteres isora Ethanol Extract Induces Apoptotic Cell Death in HepG2 Cells

Background: H. isora is a species of the genus Helicteres, a medicinal plant known to be used in the treatment of liver cancer in Vietnam. However, there has been no recorded about biological activity of H. isora species in Vietnam. The current study aimed to study the antitumor activity through its antioxidant activity and inhibitory properties on hepatocellular carcinoma cells. Methods: Arial parts of Helicteres isora were collected from Dak Nong province. Samples were extracted maceration method in ethanol solvent. The antitumor activity were identified though the DPPH radical scavenging activity (Brand-Williams et al., (1995) and WST-1 assay, Real- time PCR, Hoechst 33258 spectrofluorometric assay and Annexin V-FITC Apoptosis Assay on the human liver cancer cells (HepG2) and non-malignant cells of Chang’s Live. Result: The ethanol extract of H. isora exhibited antioxidant activity with IC50=21.16 µg/ml. H. isora extract also exhibited inhibitory activity on HepG2 cell division with IC50=37 µg/ml after 48 h. The expression of DNA fragmentation of the extract also increased with time (48-72h). Apoptosis induction of extract was associated with up-regulation of pro-apoptotic Bax gene expression and down-regulation of anti-apoptotic Bcl-2 gene expression. The H. isora extract was found to induce the apoptotic process in HepG2 cells through the up and down regulation of the Bax/Bcl-2 gene.

Apigenin promotes cell death in NCI-H23 cells by upregulation of PTEN: potential involvement of the binding of apigenin with WWP2 protein

The tumour suppressor protein PTEN is often down-regulated in non-small cell lung cancer. A major protein promoting the lowering of the PTEN protein is WWP2. Polyphenols have been shown to promote the expression of tumour suppressor genes like PTEN. We carry out the study to check for the ability of apigenin to bind with the WWP2 protein using in-silico investigation comprising docking and simulation. We checked for the cytotoxic effect of apigenin upon the non-small cell lung cancer cell line NCI-H23. We checked the PTEN expression status at the gene and protein levels. The expression levels of the apoptotic regulators BCL2, BAX and CASPASE3 genes along with the activity levels of the caspase-3 protein were checked. The ultrastructure of the cells was analysed. Our Autodock analysis showed that apigenin bound favourably with the WWP2 protein. Molecular dynamics simulation revealed that apigenin increased the parameters of RMSD, Rg and SASA when bound with the WWP2 protein. The protein-ligand complex had hydrogen bonding and majorly van der Wal’s interactions. PCA analysis revealed greater fluctuations in the apigenin-bound state of the protein. The mutant form of the WWP2 revealed similar results in the presence of apigenin. Apigenin showed efficacy against the NCI-H23 cell line and promoted PTEN protein levels, lowered BCL2 gene expression and up-regulated BAX and CASPASE3 gene expression. Increased caspase-3 activity and ultra-structural analysis revealed the occurrence of apoptosis. Thus the binding of apigenin with WWP2 could promote PTEN protein levels and lead to apoptotic activity in NCI-H23 cells. Communicated by Ramaswamy H. Sarma

Protective effect of a standardized Allium jesdianum extract in an Alzheimer's disease induced rat model

Alzheimer's disease (AD) is a complex disorder with multiple underlying mechanisms. Existing treatment options mostly address symptom management and are associated with numerous side effects. Therefore, exploring alternative therapeutic agents derived from medicinal plants, which contain various bioactive compounds with diverse pharmacological effects, holds promise for AD treatment. This study aims to assess the protective effects of the hydroalcoholic extract of Allium jesdianum on cognitive dysfunction, mitochondrial and cellular parameters, as well as genetic parameters in an intracerebroventricular Streptozotocin (icv-STZ) induced rat model of AD. Male Wistar rats were injected with a single dose of STZ (3 mg/kg, icv) to establish a sporadic AD model. A. jesdianum extract (100, 200, and 400 mg/kg/day) and donepezil (5 mg/kg/day) were orally administered for 14 days following model induction. Cognitive function was evaluated using the radial arm water maze test. Mitochondrial toxicity parameters in various brain regions (whole brain, frontal cortex, hippocampus, and cerebellum) were assessed. Gene expression analysis of miR-330, miR-132, Bax, and Bcl-2 in isolated rat brain neurons was performed using RT-qPCR. A. jesdianum extract significantly attenuated cognitive dysfunction and mitigated mitochondrial toxicity induced by icv-STZ administration. Following STZ injection, there was upregulation of Bax gene expression and downregulation of miR-330, miR-132, and Bcl-2 gene expression. Treatment with A. jesdianum extract resulted in the reversal of the expression of these microRNAs and genes, indicating its potential for improving AD and reducing neuronal apoptosis. This study demonstrates the neuroprotective capabilities of A. jesdianum against STZ-induced oxidative stress and cognitive impairment in rats, highlighting its therapeutic potential in the management of AD.

Genotoxic effect of Tamiflu and Adamine on DNA content in male albino mice

BackgroundTamiflu (Oseltamivir) and Adamine (Amantadine HCl) are antiviral drugs which are used for prevention and treatment for influenza. The present study was carried out to evaluate the effect of Tamiflu and Adamine on DNA content of hepatocytes of adult male albino mice.ResultsThe results of comet assay showed that treatment with Tamiflu and Adamine significantly increased (P < 0.005) the mean of DNA damage in hepatocytes of the treated mice in time dependent manner compared to control one. However qRT-PCR analysis showed the mRNA expression of BAX in mice liver of all treated groups with Adamine was significantly increased (P < 0.05) after ten days of treatment while the same gene was significantly decreased in Tamiflu treated group after ten days compared to control one, While the treatment of mice with both drugs simultaneously showed a significant increase (P < 0.05) in expression of BCL-2 gene and a decrease in expression of BAX gene after fifteen days of treatment.ConclusionTamiflu and Adamine had produced deleterious impacts on DNA content of hepatocytes of treated mice. Whereas, the results of treatments with Adamine indicated pathogenic impacts more than that induced by Tamiflu.

Human Umbilical Cord Mesenchymal Stem Cell-Derived Microvesicles Could Induce Apoptosis and Autophagy in Acute Myeloid Leukemia.

Microvesicles (MV) have been identified as candidate biomarkers for treating acute myeloid leukemia (AML). This study investigated the effects of human umbilical cord-derived mesenchymal stem cell (hUCMSC)-derived MVs on apoptosis and autophagy in the KG-1 leukemic cell line. The hUCMSCs were cultured and characterized by flow cytometry. MVs were isolated by ultracentrifugation, and the concentration was determined using the Bradford method. The characteristics of MVs were confirmed using transmission electron microscopy, flow cytometry, and dynamic light scattering methods. KG-1 cells were treated with the desired concentrations of MVs for 24 h. The apoptosis induction and reactive oxygen species production were evaluated using flow cytometry. RT-PCR was performed to evaluate apoptosis- and autophagy-related genes expression. Following tretment of KG-1 cells with 25, 50, and 100 μg/ml concentrations of MVs, the apoptosis rates were 47.85%, 47.15%, and 51.35% (p < 0.0001), and the autophagy-induced ROS levels were 73.9% (p < 0.0002), 84.8% (p < 0.0001), and 85.4% (p < 0.0001), respectively. BAX and ATG7 gene expression increased significantly at all concentrations compared to the control, and this level was higher at 50 μg/ml than that of the other concentrations. In addition, LC3 and Beclin 1 expression increased significantly in a concentration-dependen manner. Conversely, BCL2 expression decreased compared to the control. Our findings indicate that hUCMSC-MVs could induce cell death pathways of autophagy and apoptosis in the KG-1 cell lines and exert potent antiproliferative and proapoptotic effects on KG-1 cells in vitro. Therefore, hUCMSC-MVs may be a potential approach for cancer therapy as a novel cell-to-cell communication strategy.

Evaluation of Apoptosis-Related Genes and Hormone Secretion Profiles Using Three Dimensional Culture System of Human Testicular Organoids

Background: In reproductive biology, testicular organoids can be used to treat infertility and to study testicular development and spermatogonial stem cells (SSCs) differentiation. Generating organoid from primary cells is challenging. In this study, testicular organoids were created using human primary testicular cells and evaluated the apoptotic gene expression and hormone secretion profiles of the organoids. Materials and Methods: Primary human testicular cells were isolated using 2-step enzymatic digestion from three brain-dead donors. Immunocytochemistry and flow cytometry analyses were performed to confirm human SSCs. Isolated cells were cultured in three experimental groups: control group (2 dimensional (2D)), group 1 (organoid culture after 2D culture), and group 2 (organoid culture immediately after enzymatic digestion). Testicular organoids were cultured in DMEM/F-12 media supplemented with follicle-stimulating hormone (FSH) and fetal bovine serum (FBS) for four weeks. After 24 hours and four weeks of culture, reverse transcription quantitative real-time PCR (RT-qPCR) was used to investigate the relative expression of apoptotic genes (caspase 3, 9, Bax, and Bcl-2). At 24 hours, two weeks, and four weeks after culture, enzyme-linked immunoassay (ELISA) was used to determine the testosterone and inhibin B concentrations. Light microscopy and toluidine blue staining were also used for morphological analysis. Results: RT-qPCR results revealed that pro-apoptotic (caspase 3, 9, Bax) gene expression levels were highest in group 2 after 24 h and four weeks of culture. In contrast, the expression level of Bcl-2 (anti-apoptotic) was lower in group 2 compared to other groups. The hormone secretion levels decreased in a time-dependent manner during the cultivation. According to morphological evaluations, testicular organoids are compact, spherical structures with two to three elongated cells organized along their border. Conclusion: Our findings revealed that the testicular organoid culture system maintained hormonal secretory abilities, demonstrating the function of Sertoli and Leydig cells in the absence of testis-specific environments.

Evaluation of the cytotoxic effect of GaFe2O4@Ag magnetic nanocomposite biosynthesized by Spirulina platensis on breast cancer cell line and evaluation of BAX, Bcl-2, CASP9 and MMP1 genes expression

Treating breast cancer is hardly achieved and finding efficient anticancer nanocompounds has gained great attention. In this work, the cytotoxic effect of GaFe2O4@Ag nanocomposite biosynthesized by Spirulina platensis on breast cancer cell line, and expression of the BAX, Bcl-2, CASP9 and MMP1 (Matrix Metallopeptidase 1) genes were evaluated. Physicochemical features of the nanocomposite were determined using the FT-IR, XRD, SEM, TEM, EDX-mapping, VSM, Zeta potential, and DLS analyses. The cytotoxic effect of the nanocomposite for MCF-7 and HEK-293 cells was evaluated by the MTT assay. Flow cytometry analysis, Caspase-3 activation assay, and Hoechst staining were performed to evaluate the apoptosis induction potential of the nanocomposite. Further, the relative expression of the Bcl-2, BAX, and CASP9, and MMP1 genes was determined by quantitative PCR assay. The prepared nanocomposite was spherical with a size range of 35–60 nm. The hydrodynamic size and zeta potential of the nanocomposite were 328 nm and −31.8 mv, respectively. GaFe2O4@Ag nanocomposite had a higher cytotoxic effect on breast cancer cells than normal human cells with the IC50 of 18.6 and 220 µg/mL, respectively. Treating breast cancer cells with the nanocomposite induced apoptosis among 85.2% of cells, increased caspase-3 activity by 4.3 folds, and caused apoptotic nuclear changes. Also, GaFe2O4@Ag reduced the expression of the Bcl-2 and MMP1 by 1.3 and 0.6 folds and up-regulated the BAX and CASP9 genes by 2.7 and 2.65 folds, respectively. Our results revealed that GaFe2O4@Ag was highly cytotoxic for breast cancer cells via triggering apoptosis pathways and could be considered as a novel and efficient agent against breast cancer, after further in-vivo experiments.

Anticancer Activity of Postbiotic Mediators Derived from Lactobacillus Rhamnosus GG and Lactobacillus Reuteri on Acute Lymphoblastic Leukemia Cells.

Leukemia remains a global health challenge, requiring the exploration of alternative therapies with reduced side effects. Probiotics, particularly Lactobacillus species, have gained attention because of their potential anticancer properties. This study investigated the anticancer and cytotoxic effects of postbiotic mediators (PMs) derived from Lactobacillus rhamnosus GG (LGG) and Lactobacillus reuteri (LR) on acute lymphoblastic leukemia (ALL) cells and peripheral blood mononuclear cells (PBMCs). The PMs were prepared by culturing LGG and LR strains and isolating the supernatant. The MTT assay assessed cell viability on ALL Jurkat cells and PBMCs, and apoptosis analysis was conducted using flow cytometry. Quantitative real-time PCR was also performed to analyze BAX, BCL-2, BCLX, FAS, and p27 gene expression levels. The results showed that PMs derived from LGG and LR significantly reduced cell viability in Jurkat cells (P0.05) but not PBMCs (P0.05). Apoptosis analysis revealed an increase in apoptotic cells after PMs treatment. Nevertheless, gene expression analysis revealed no statistically significant difference between the treated and untreated groups in BAX, BCL-2, BCLX, FAS, and p27 gene expression levels (P0.05). Findings suggest that specific PMs derived from LGG and LR possess anticancer properties against ALL cells. This research highlighted the promise of PMs as a cutting-edge and less toxic adjuvant therapeutic strategy in cancer treatment.

Fabricating niosomal-PEG nanoparticles co-loaded with metformin and silibinin for effective treatment of human lung cancer cells.

Despite current therapies, lung cancer remains a global issue and requires the creation of novel treatment methods. Recent research has shown that biguanides such as metformin (MET) and silibinin (SIL) have a potential anticancer effect. As a consequence, the effectiveness of MET and SIL in combination against lung cancer cells was investigated in this study to develop an effective and novel treatment method. Niosomal nanoparticles were synthesized via the thin-film hydration method, and field emission scanning electron microscopy (FE-SEM), Fourier transform infrared (FTIR), atomic force microscopy (AFM), and dynamic light scattering (DLS) techniques were used to evaluate their physico-chemical characteristics. The cytotoxic effects of free and drug-loaded nanoparticles (NPs), as well as their combination, on A549 cells were assessed using the MTT assay. An apoptosis test was used while under the influence of medication to identify the molecular mechanisms behind programmed cell death. With the use of a cell cycle test, it was determined whether pharmaceutical effects caused the cell cycle to stop progressing. Additionally, the qRT-PCR technique was used to evaluate the levels of hTERT, BAX, and BCL-2 gene expression after 48-h medication treatment. In the cytotoxicity assay, the growth of A549 lung cancer cells was inhibited by both MET and SIL. Compared to the individual therapies, the combination of MET and SIL dramatically and synergistically decreased the IC50 values of MET and SIL in lung cancer cells. Furthermore, the combination of MET and SIL produced lower IC50 values and a better anti-proliferative effect on A549 lung cancer cells. Real-time PCR results showed that the expression levels of hTERT and BCL-2 were significantly reduced in lung cancer cell lines treated with MET and SIL compared to single treatments (p< 0.001). It is anticipated that the use of nano-niosomal-formed MET and SIL would improve lung cancer treatment outcomes and improve the therapeutic efficiency of lung cancer cells.

Forskolin Improves Male Reproductive Complications Caused by Hyperglycemia in Type 2 Diabetic Rats.

In many diabetic patients, spermatogenesis complications are frequent causing infertility problems. This study aimed to demonstrate the effect of Forskolin on male reproductive dysfunction caused by type 2 diabetes. In this experimental study, type 2 diabetes was induced by a high-fat diet (HFD) for one month and then a low single dose injection (35 mg/kg) of streptozotocin (STZ) in Wistar rats. After 72 hours, rats with more than 200 mg/dl of blood glucose were considered type 2 diabetic rats. Forty rats (200-250 g) were divided into four groups (n=10) including group 1 (G1): rats with normal diet and buffer citrate (STZ solvent) injection, group 2 (G2): control type 2 diabetic rats with HFD and STZ injection, group 3 (G3): type 2 diabetic rats received phosphate buffer saline (PBS) as Forskolin solvent, and group 4 (G4): Forskolin treated diabetic rats (10 mg/kg) for 1 month. In comparison to control group, in diabetic groups (G2 and G3) some parameters are increased significantly: The blood glucose (P=0.00078), testicular malondialdehyde (MDA) level and body weight (P=0.00009) and Bax gene expression (P=0.00007). Unlike, some parameters are decreased significantly: The serum level of testosterone (P=0.0009), testicular superoxide dismutase (SOD, P=0.00007) and glutathione peroxidase (GPX) levels (P=0.00008), sperm concentration (P=0.00008), motility (P=0.00009), normal morphological sperm (P=0.00008) and Bcl-2 gene expression (P=0.00009). However, in Forskolin treated group (G4) the parameters stayed close to control values that was significantly (P=0.00007) higher than in G2 and G3 groups. Therefore, treatment with Forskolin significantly improved these abnormal changes in Forskolin-treated group. Our study demonstrates that Forskolin is an effective antidiabetic agent, which significantly improves sperm concentration, testosterone levels, and antioxidant activity in diabetic rats.

Concomitant effects of paclitaxel and celecoxib on genes involved in apoptosis of triple-negative metastatic breast cancer cells.

Although triple-negative breast cancer accounts for less than one-fifth of breast cancers, it has a higher rate of metastasis and mortality. This study investigated the effects of combination treatment with paclitaxel and celecoxib on the expression of genes involved in the apoptosis of triple-negative metastatic breast cancer cells. MDA-MB-231 cells were cultured and then treated with certain concentrations of celecoxib (CLX), paclitaxel (PTX), and combination of them for 24 and 48 h. Cell viability was assessed by the MTT method. The real-time PCR method was utilized to assess the expression level of the genes involved in apoptosis. Western blotting was used for evaluating protein expression. IC50 values for CLX and PTX were 73.95 μM and 3.15 μM, respectively. The results demonstrated that PTX, CLX, and PTX+CLX significantly (p<0.05) reduced cell viability. The comparison of combination treatment with PTX showed a significant increase in caspase 3 gene expression at both time points, in Bax gene expression after 48 h, and a remarkable decrease in Bcl-2 gene expression at both times. Western blotting results were in line with genes' expression. These findings indicate that a combination of PTX and CLX results in a significantly more reduction in cell viability of breast cancer cells. In addition, it seems CLX may be an effective agent in regulating the expression level of caspase 3, Bax, and Bcl-2 when combined with PTX.

Synergistic Effects of Capsaicin and Quercetin Improved Induced Premature Ovarian Failure in Rat.

Premature ovarian failure (POF) is a heterogeneous disorder. POF is defined as hypergonadotropic hypoestrogenism in women under 40 years. There is no effective treatment to cure POF patients. Antioxidants prevent ovarian damage by reducing the lipid peroxidation cascades affecting folliculogenesis, meiosis and ovulation. Hence; the aim of present study was to investigate the effects of Capsaicin (CAP) and Quercetin (QUR) on cyclophosphamide (CYC)-induced POF in rat model. In this experimental study, POF was induced by intraperitoneal injection of 200 mg/kg CYC on first day and then 8 mg/kg/day for the following 3 days. After 4 days of CYC administration, rats were randomly divided into five groups (n=6/group) as follows: POF, dimethyl sulfoxide (DMSO), CAP (0.5 mg/kg/day), QUR (100 mg/kg/day) and CAP+QUR. Biochemical, hormonal, gene expression, and histological evaluations were performed on blood serum and tissue samples after 14 days of treatment with the CAP and QUR. CAP, QUR and CAP+QUR groups showed signs of restored ovarian function in the form of a significant increase in serum total antioxidant capacity (TAC), estrogen, progesterone and anti-mullerian hormone (AMH) levels versus POF and DMSO groups and a significant improvement in histological parameters and follicle numbers in treatment groups compared to POF and DMSO groups. Polymerase chain reaction (PCR) analysis demonstrated that CAP and QUR upregulate the expression of BAX gene and decreased the expression of apoptosis inducing genes (BCL-2 and P53). CAP and QUR treatment of CYC-induced POF rats showed a positive effect on reducing ovarian damage by improving TAC levels, expression of apoptotic genes, levels of ovarian reserve markers, and histological parameters. Our results suggest that treatment with CAP or QUR may be a conservative treatment approach for CYC -induced POF.

Examining the Effect of Extract of Oak Fruit Jaft on AGS Cell Lines

Background: Gastrointestinal carcinoma comprises 5% of all pediatric cancer in children. Given that the possible and beneficial effect of the Jaft extract in the treatment of gastric cancer is not known and there is no comprehensive study in this regard, this study aimed to assess the effect of Jaft extract on gastric cancer cell lines.&#x0D; Materials and Methods: In this case-control study, oak fruit was collected from the mountains of Lorestan province. A gastric cancer (AGS) cell line was obtained from the Institute Pasteur cell bank and was cultured. After the preparation of the ethanolic extract of Jaft, the cell viability of the gastric cancer cells treated with Jaft extract was investigated by MTT assay. Quantitative Reverse Transcription PCR was used for assessing the expression of BAX, and BCL2 genes.&#x0D; Results: The half maximal inhibitory concentration (IC50) value of Jaft extract was 162 µg/ml. The BAX gene expression was different between the case and control groups. In this regard, the expression of the BAX gene was increased in the concentration of 162 (P&lt;0.01) and 250 µg/ml (P&lt;0.001) of Jaft extract compared to the control group. The BCL2 expression was different between the two groups (P&lt;0.05). In this regard, the expression of the BCL2 gene was decreased in the concentration of 162 and 250 µg/ml of Jaft extract compared to the control group.&#x0D; Conclusion: It was found that Jaft extract increased the apoptosis of gastric cancer cells; therefore, it seems that the hydroalchoholic extract of Jaft is an appropriate anticancer medication.

Melissa officinalis extract suppresses endoplasmic reticulum stress-induced apoptosis in the brain of hypothyroidism-induced rats exposed to γ-radiation

The purpose of this study was to demonstrate the neuroprotective effect of Melissa officinalis extract (MEE) against brain damage associated with hypothyroidism induced by propylthiouracil (PTU) and/or γ-radiation (IR) in rats. Hypothyroidism induction and/or exposure to IR resulted in a significant decrease in the serum levels of T3 and T4 associated with increased levels of lipid peroxidation end product, malondialdehyde (MDA), and nitrites (NO) in the brain tissue homogenate. Also, hypothyroidism and /or exposure to IR markedly enhance the endoplasmic reticulum stress by upregulating the gene expressions of the protein kinase RNA-like endoplasmic reticulum kinase (PERK), activated transcription factor 6 (ATF6), endoplasmic reticulum-associated degradation (ERAD), and CCAAT/enhancer-binding protein homologous protein (CHOP) in the brain tissue homogenate associated with a proapoptotic state which indicated by the overexpression of Bax, BCl2, and caspase-12 that culminates in brain damage. Meanwhile, the PTU and /or IR-exposed rats treated with MEE reduced oxidative stress and ERAD through ATF6. Also, the MEE treatment prevented the Bax and caspase-12 gene expression from increasing. This treatment in hypothyroid animals was associated with neuronal protection as indicated by the downregulation in the gene expressions of the microtubule-associated protein tau (MAPT) and amyloid precursor protein (APP) in the brain tissue. Furthermore, the administration of MEE ameliorates the histological structure of brain tissue. In conclusion, MEE might prevent hypothyroidism-induced brain damage associated with oxidative stress and endoplasmic reticulum stress.

Ceratonia siliqua pods (Carob) methanol extract alleviates doxorubicin-induced nephrotoxicity via antioxidant, anti-inflammatory and anti-apoptotic pathways in rats

Doxorubicin (DOX) is an anti-neoplastic therapy, but its use is limited by its deleterious toxic effects including nephrotoxicity and cardiotoxicity. This work aimed at assessing the potential protective effect of Ceratonia siliqua methanol extract (CME) on DOX-induced nephrotoxicity in 5 groups of Wistar rats. Nephrotoxicity was induced experimentally by intraperitoneal (IP) injection of DOX (15 mg/kg). DOX increased serum creatinine, urea, sodium, and potassium levels. It elevated MDA levels in the renal tissue but decreased the concentration of GSH and the activity of GST, CAT, and SOD. Meanwhile, it decreased the level of immunomodulatory anti-inflammatory mediators: IL-10 and TGF-β, as well as the activity of MPO but increased the level of IL-6, TNF-α, and caspase-3 in the renal tissue. DOX has upregulated COX-2, caspase-9, and Bax gene expression and downregulated the Bcl-2 gene expression. Immunolabeling of renal tubular epithelium in DOX-intoxicated rats was moderate to strong against Bax, COX-2, and NF-kβ and weak against Bcl-2. Treatment with CME significantly restored the levels of kidney function parameters and the levels of oxidative stress markers. It stimulated the production of IL-10 and TGF-β and decreased the level of IL-6 and TNF-α. CME reverted the gene expression of COX-2, caspase-9, and Bax. Microscopically, CME alleviated the DOX-induced renal damage. Phytochemical analysis revealed the presence of 26 compounds in the CME. No signs of acute toxicity were recorded by CME up to 4000 mg/kg b. wt. orally into mice. Finally, CME could effectively alleviate the deleterious effects of DOX on the kidney. The safety of carob extract encourages its use in the preparation of valuable therapeutic agents.