Bax Gene Expression Research Articles

Neuroprotective effects of tranexamic acid against hydrogen peroxide-induced cytotoxicity on human neuroblastoma SH-SY5Y cells

ABSTRACT Introduction Tranexamic acid (TA) is an anticoagulant drug that is used worldwide. However, the adverse effects of TA may insult the nervous system. This study aimed to investigate the dual effects of TA on SH-SY5Y cells, including its detrimental and neuroprotective effects. Methods SH-SY5Y cells were treated with various concentrations of TA and exposed to H2O2 for 24 hours. The neuroprotective effects of TA were evaluated in H2O2-challenged cells. To assess the neuroprotective effects of TA, SH-SY5Y cells were pretreated with TA for 12 hours and then exposed to H2O2 for 24 hours. Cell viability was assessed using the MTT assay. Flow cytometry was used to evaluate cellular apoptosis. The expression of Bax, Bcl-2, and Caspase-3 genes was analyzed by real-time PCR. Additionally, Akt phosphorylation was evaluated using western blotting. Results At high concentrations, TA reduced cell viability and induced apoptosis by up-regulating BAX and Caspase-3 gene expression and down-regulating BCL-2 transcript. Furthermore, Akt phosphorylation was reduced following TA treatment. TA exhibited protective effects against H2O2-induced cell stress by down-regulating Bax and Caspase-3 gene expression, up-regulating Bcl-2 expression, and increasing the p-AKT/AKT ratio. Conclusion Our findings demonstrated that TA exerts its neuroprotective effect at lower concentrations, but induces apoptosis in SH-SY5Y cells at high concentrations.

Livogrit mitigates ANIT-induced cholestasis-like symptoms in an in vivo model by curbing hepatic inflammation and regulating BAX, TGF-β, MMP-9 and α-SMA gene expression

Livogrit mitigates ANIT-induced cholestasis-like symptoms in an in vivo model by curbing hepatic inflammation and regulating BAX, TGF-β, MMP-9 and α-SMA gene expression

Curcumin chemo-sensitizes intrinsic apoptosis through ROS-mediated mitochondrial hyperpolarization and DNA damage in breast cancer cells.

Curcumin (CUR), a polyphenol phyto-compound extracted from turmeric rhizome (Curcuma longa), suppresses cancer by inducing apoptosis while also limiting cell survival and proliferation. This in vitro and in silico research work focuses on the synergistic sensitization of Doxorubicin (DOXO) on regulating ROS-mediated apoptosis of the breast cancer cells (MDA-MB-231 and MCF-7) in DOXO-CUR co-treatment. We observed dose-dependent cytotoxicity, increased ROS production, and mtDNA fragmentation by reduced membrane potential. The combined molecular docking of Bcl2, Bax, and Caspase3 proteins with DOXO and CUR with lower binding energies proves the stable interactions of protein-ligand complexes in the combination doses. Cell survival was measured by MTT and flow cytometry assays. Mitochondrial ROS production, mtDNA condensation, and MMP depletion were documented using fluorescence micrographs. The enrichment analysis of the ROS pathway genes by RT-qPCR (relative fold change) indicates the activation of Caspase3-mediated intrinsic apoptosis. Autodock 4.2 and Gromac 2022.4 were performed for in silico binding interaction and stability analysis. Our study calculates the DOXO and CUR combination (0.33 + 33 μM in MDA-MB-231 and 0.14 + 14 μM in MCF-7) shows maximum growth inhibition (70-75 %) by elevated oxidative stress and reduced membrane potential, which suggests that CUR could be a potential therapeutic agent for treating breast cancers in near future. The method of apoptosis was further analyzed, where elevated cellular ROS level by CUR + DOXO combination therapy depleted mitochondrial membrane potential and enhanced the DNA condensation. The mitochondrial pro-apoptotic genes BAX, BAK, BIM, CASPASE9, and CASPASE3 and anti-apoptotic BCL2 gene expressions depicted triggering intrinsic apoptosis pathway, co-relating with the in silico molecular docking, simulation, and MM-PBSA energy calculations. The synergism between CUR and DOXO was also validated by increased binding affinity and reduced inhibitory constant against key proteins Bcl2, Bax, and Caspase3. Bcl2-DOXO showed BE: -5.03 and Bcl2-DOXO-CUR showed BE: -4.7. Whereas, Bax-DOXO binding energy was -5.49 and Bax-DOXO-CUR binding was -3.83. The most preferable synergistic binding was found with Caspase3 protein, where Caspase3-DOXO docking energy was -1.63 but Caspase3-DOXO-CUR combined docking energy was -3.51. The stability of protein-ligand complexes was accessed with MD simulations and binding free energy calculations, Bcl2-CUR-DOXO combination complex showed ∆G: -25.62, Bax-DOXO-CUR complex showed the maximum ∆G: -34.18, with Caspase-CUR-DOXO complex (∆G: -15.18), indicating the proteins most stable conformation while interacting with CUR + DOXO combination. The correlation between the in vitro and in silico analysis of apoptosis pathway components widens the further research scope for proteomics and organoid studies, which might be worthy for successful clinical trials.

Fluoxetine Mitigates Human Sperm Quality by Disrupting the Antioxidant Defense System and Altering the Expression of Apoptosis-Related Genes: An In Vitro Study.

Fluoxetine is used in the management of depression, anxiety and other mood disorders by increasing serotonin levels in the brain and can cause sexual side effects by changing the homeostasis of sex hormones and increasing oxidative stress. Since many men who take fluoxetine are of reproductive age and sperm are exposed to fluoxetine for a considerable time, this study aimed to examine the in vitro effects of fluoxetine on human sperm biochemical markers and sperm parameters. Semen samples from 30 fertile men were divided into three groups: a positive control group, a negative control group and a fluoxetine-treated group. We investigated sperm parameters, mitochondrial membrane potential (MMP), acrosome reaction, DNA fragmentation, chromatin integrity, and the expression of CASPASE8, CASPASE9, BAX, and BCL2 genes and proteins. Data were analyzed using repeated measures analysis. The results showed that the average percentage of motility, viability, MMP, acrosome and chromatin integrity, total antioxidant capacity (TAC) level, and BCL2 gene and protein expression in the fluoxetine group were significantly reduced compared to the positive and negative control groups. While the average percentage of non-progressive motility, sperm DNA fragmentation, malondialdehyde (MDA) level, reactive oxygen species (ROS), gene and proteins expression of CASPASE8, CASPASE9 and BAX increased significantly. This study suggests that fluoxetine may impair sperm quality by increasing the expression of apoptotic genes, proteins, and oxidative stress. Therefore, careful management of fluoxetine in treating depression is crucial, especially in men of reproductive age, due to its potential sexual side effects. HIGHLIGHTS: • Fluoxetine reduces the quality of human sperm by inducing oxidative stress. • Fluoxetine lowers the total antioxidant capacity in human sperm by increasing ROS. • Fluoxetine increases the expression of apoptosis genes in human sperm.

Limonene as a potential cardioprotective agent: mechanistic intuitions into antioxidant and anti-apoptotic properties in isoproterenol-induced ischemia in rats

Background and aims: Excessive oxidative stress and increased apoptosis are the main culprits to heart damage associated with ischemic heart disease. This study aimed to demonstrate the cardioprotective properties of limonene (LIM), a monoterpenoid compound, in isoproterenol-induced ischemia (ISO-I) in rats. Methods: Forty male Wistar rats were divided into five groups (n=8). Normal saline (1 mL/kg) and LIM at doses of 10 mg/kg, 20 mg/kg, and 40 mg/kg were administered for seven days before the induction of ischemia by ISO. Creatine kinase-MB (CK-MB), creatine phosphokinase (CPK), lactate dehydrogenase (LDH), malondialdehyde (MDA), total antioxidant capability (TAC), and relative expression of BCL2 and BAX genes underwent measurement. Results: CK-MB (3.73±0.560 versus 0.888±0.317), CPK (901±72.2 versus 397±33.5), LDH (1992±176 versus 821±94.1), and MDA levels in the serum (65.6±11.0 versus 28.3±4.75) and heart (104±8.70 versus 65.3±7.46) samples significantly increased following ischemia in the treatment groups compared to the control group (P<0.001 for all markers). Ischemia decreased TAC in the heart (324±45.0 versus 588±64.5) and serum (202±23.0 versus 388±45.9) samples compared to the control counterparts (P<0.001 for both samples). ISO-I is associated with a decrease in Bcl-2 (0.880±0.118 versus 1.74±0.178) and an increase in BAX (2.75±0.124 versus 1.08±0.0938) gene expression in comparison to the control group (P<0.001 for both genes). LIM could reverse the aforementioned heart damage-related biomarkers in the serum and heart samples. Finally, LIM improved histopathological disarrangement following ischemia. Conclusion: The findings revealed that LIM, at least partially due to its antioxidant and anti-apoptotic potentials, exerted cardioprotection against ISO-induced heart ischemia in rats.

Alterations in the expression of genes involved in the mitochondrial apoptotic pathway upon exposure to lead oxide nanoparticles

Introduction. Lead production technologies pollute the air with aerosol nanoparticles, including those of lead oxide (PbO NPs). Lead can cause oxidative stress that leads to cell death. Experimental studies of effects of PbO NPs at the gene transcription level will expand our knowledge of the mechanisms of PbO NP toxicity and improve assessment of health risks for the population exposed to them. The purpose was to study the expression of genes involved in antioxidant protection and apoptosis following subchronic inhalation exposure of lead nanoparticles to rats. Materials and methods. Female albino rats were exposed to PbO NPs in an inhalation chamber at a concentration of 1.55 ± 0.06 mg/m3, 4 hours a day, 5 days a week for 1 month; the control group breathed clean air in a similar chamber. After exposure cessation, RNA was isolated from fragments of the olfactory bulb, cerebellum, lung, and liver. Expression of the P53, BAX, BCL-2, GSTM1, GSTP1, and SOD2 genes was determined by quantitative real-time PCR. The data was analyzed using the Mann-Whitney test. Results. In the olfactory bulb, BCL-2 gene expression was significantly lower, while that of P53 was higher in the exposed rodents compared to the controls. In the cerebellum of the exposed animals, BAX and P53 genes expression was statistically higher and lower than in the control group, respectively. BCL-2 gene expression in the liver was significantly lower in the exposed group. Limitations. The experiment involved only female rats, so it does not take into account sex differences and considers only gene expression, neglecting post-translational mechanisms and protein expression. Conclusion. Inhalation exposure to PbO NPs at the concentration of 1.55 ± 0.06 mg/m3 causes changes in the expression of genes associated with mitochondrial apoptosis in the brain and liver, but not in the lungs of laboratory rats.

Protective effects of soybean peptides on H2O2-induced oxidative injury in IPEC-J2 cells

The purpose of the study was to demonstrate how soybean peptides (SBP) protect against H2O2 -induced injury in intestinal porcine epithelial cells (IPEC-J2). SBP were prepared by protease hydrolysis, in which the molecular weights of 95.76% SBP were smaller than 3 kDa. Cell experiment included four groups: Control group (IPEC-J2 cells were treated with HGDMEM), SBP group (100 μg/mL SBP incubation for 13 h), H2O2 treatment group (1 mM H2O2 treatment for 1 h), SBP + H2O2 group (100 μg/mL SBP pretreatment for 12 h followed by 1 mM H2O2 treatment for 1 h). This study showed that that treatment with single 1 mM H2O2 for 1 h significantly reduced cell viability to 52.99% (p < 0.05), up-regulated Bax and Caspase-3 gene expressions (p < 0.05), and down-regulated gene expressions of ZO-1, CAT, SOD1, HO-1 and Nrf2 (p < 0.05), compared with the control group. However, pretreatment with SBP followed by H2O2 inducement significantly increased cell viability to 72.99%, decreased cell apoptosis, increased SOD, CAT and GSH-Px activity (p < 0.05), down-regulated Bax and Caspase-3 gene expressions (p < 0.05), and up-regulated the gene expressions of ZO-1, Claudin-1, Occludin, catalase, glutathione GPX1, SOD1, HO-1, NQO1 and Nrf2, compared with the single H2O2–induced cells. According to the study, SBP pretreatment reduced H2O2-induced oxidative stress in cells and preserved the integrity of intestinal cells.

Antitumor Activity and Immunostimulant Properties of Liposomes Containing Rosemary Extract on a Mouse Model of Colorectal Cancer.

Rosemary (Ros) is a member of the Lamiaceae family known for its antitumor properties. However, its low water solubility and impaired bioavailability are limiting factors when using rosemary extract. Liposomes are synthetic vesicles that offer permeability, improved bioavailability, and lack of immunogenicity and toxicity, making them ideal for delivering various drugs. To prepare liposomes (HSPC/Chol/mPEG2000-DSPE) containing rosemary alcoholic extract (LipRos) and evaluate its antitumor properties in a mouse model of colorectal cancer (CRC). LipRos were prepared and characterized. CRC was induced in Balb/c mice by subcutaneous injection of C26 cells, and tumor size was monitored continuously. The MTT assay was performed to evaluate cytotoxicity, and liver and kidney function tests were conducted to assess safety. The expression of the pro-apoptotic gene B-cell-lymphoma-2 (Bcl-2), the anti-apoptotic gene Bcl-2-associated-X-protein (Bax), and the expression of cytokines Tumor-necrosis-factor-alpha (TNF-α), Transforming-growth-factor-beta (TGF-β), and Interferon-gamma (IFN-γ) were investigated using real-time PCR. Flow cytometry was used to evaluate the count of cytotoxic (CTL) and regulatory T lymphocytes (Tregs) in spleen and tumor tissue. The results showed that the size of liposomal formulations and their encapsulation efficiency were 113.4 nm and 85%, respectively. The MTT assay demonstrated insignificant cytotoxicity of LipRos on splenocytes, and the tumor size was significantly reduced in the LipRos group (P=0.00045). LipRos also significantly decreased Bcl-2 gene expression (P=0.0086), increased Bax and IFN-γ gene expression (P=0.031), and enhanced the infiltration of CTLs in tumor tissue (P=0.023). This study showed that PEGylated (Poly-Ethylene-Glycol) liposomes containing rosemary extract exhibit an antitumor effect on C26 colorectal cancer cells through multiple mechanisms. These findings can be utilized in future studies.

Anti-cancer effect of sodium pentaborate in combination with cisplatin on lung cancer cell lines.

Despite the development of novel therapeutic modalities, lung cancer persists as the leading cause of cancer-related mortality. Platinum-based treatments represent the most prominent treatment option, with cisplatin being the most frequently utilized chemotherapeutic agent. However, cisplatin has several serious side effects. A substantial body of evidence has emerged in recent years indicating that boron compounds exhibit anti-cancer properties when administered as monotherapy or in combination with chemotherapy agents. The objective of this study is to examine the anti-cancer effects of Cisplatin (Cis) and sodium pentaborate pentahydrate (NaB), both individually and in combination, on non-small cell lung cancer (NSCLC) cell line A-549 cells and small cell lung cancer (SCLC) cell line DMS-114 cells under in vitro conditions. The effects of cisplatin and NAB on cell survival, apoptosis, the cell cycle, and the expression levels of apoptotic, anti-apoptotic, and tumor suppressor genes were determined by an MTS assay, an Annexin-V assay, a cell cycle analysis, and real-time PCR (qPCR). It was found that the IC-50 value of cisplatin, which was 10 µM when used alone, decreased to 2.5 µM when combined with a non-toxic dose of NaB on the A-549 cell line. BAX and TP53 gene expression levels were elevated by the Nab-Cisplatin combination in the A-549 cell line. The combination was observed to result in an approximately 19-fold increase in CDK2 gene expression in the A-549 cell line and an approximately 6-fold increase in the DMS-114 line, which resulted in S phase and/or G2 phase arrest on the cell cycle. Gene expression levels of Survivin and Ki-67 were decreased by the combination on both cell lines when compared with cisplatin alone. The findings demonstrate that NaB exerts an anti-cancer effect on the A-549 and DMS-114 cell lines. Moreover, when combined with cisplatin, it produces a synergistic anti-cancer effect on the A-549 cell line, whereby apoptosis is activated and cell proliferation is inhibited. The combination of NaB and cisplatin represents a novel approach to the treatment of NSCLC. This is due to the fact that it reduces the IC-50 value of cisplatin and also results in a greater inhibition of cell division and a stronger induction of cell death when used in the context of a combined treatment. Further insight into the effects of the NaB-cisplatin combination will be gained from in vivo experiments and clinical studies.

Clemastine mitigates sepsis-induced acute kidney injury in rats; the role of α-Klotho/TLR-4/MYD-88/NF-κB/ Caspase-3/ p-P38 MAPK signaling pathways

Sepsis is a fatal condition, with an annual incidence of more than 48 million cases as well as 11 million deaths resulting from it. Moreover, sepsis continues to rank as the fifth most prevalent cause of mortality globally. The objective of this study is to investigate if Clemastine (CLM) pretreatment protects against acute kidney injury (AKI) caused by cecal ligation and puncture (CLP) via modulating Toll-like receptor-4 (TLR-4), Myeloid differentiation primary response 88 (MYD-88), nuclear factor kappa B (NF-κB), Bcl-2-associated X (Bax), B-cell lymphoma-2 (Bcl-2), and caspase-3 signaling pathways. CLM markedly attenuated sepsis-caused molecular, biochemical, and histopathological alterations. CLM downregulated the levels of the proinflammatory markers, suppressed the expression of cleaved caspase-3, TLR-4 and MYD-88 as well as inactivating NF-κB p-P65 and p-P38 proteins, inhibited Bax, NF-κB, and caspase-3 genes expression, and augmented α-Klotho protein expression as well as Bcl-2 gene expression. Finally, CLM pretreatment protected against acute kidney injury by preventing TLR-4/p-P38 pathway-mediated apoptotic cell death in rats.

P53 and the E3 Ubiquitin Ligase MDM2 in Glaucomatous Lamina Cribrosa Cells.

Lamina cribrosa (LC) cells play an integral role in extracellular matrix remodeling and fibrosis in human glaucoma. LC cells bear similarities to myofibroblasts that adopt an apoptotic-resistant, proliferative phenotype, a process linked to dysregulation of tumor suppressor-gene p53 pathways, including ubiquitin-proteasomal degradation via murine-double-minute-2 (MDM2). Here, we investigate p53 and MDM2 in glaucomatous LC cells. Primary human LC cells were isolated from glaucomatous donor eyes (GLC) and age-matched normal controls (NLC) (n = 3 donors/group). LC cells were cultured under standard conditions ± 48-h treatment with p53-MDM2-interaction inhibitor RG-7112. Markers of p53-MDM2, fibrosis, and apoptosis were analyzed by real-time polymerase chain reaction (qRT-PCR), western blotting, and immunofluorescence. Cellular proliferation and viability were assessed using colorimetric methyl-thiazolyl-tetrazolium salt assays (MTS/MTT). In GLC versus NLC cells, protein expression of p53 was significantly decreased (p < 0.05), MDM2 was significantly increased, and immunofluorescence showed reduced p53 and increased MDM2 expression in GLC nuclei. RG-7112 treatment significantly increased p53 and significantly decreased MDM2 gene and protein expression. GLC cells had significantly increased protein expression of αSMA, significantly decreased caspase-3 protein expression, and significantly increased proliferation after 96 h. RG-7112 treatment significantly decreased COL1A1 and αSMA, significantly increased BAX and caspase-3 gene expression, and significantly decreased proliferation in GLC cells. MTT-assay showed equivocal cellular viability in NLC/GLC cells with/without RG-7112 treatment. Our data suggests that proliferation and the ubiquitin-proteasomal pathway are dysregulated in GLC cells, with MDM2-led p53 protein degradation negatively impacting its protective role.

Cannabidiol mitigates methotrexate-induced hepatic injury via SIRT-1/p53 signaling and mitochondrial pathways: reduces oxidative stress and inflammation

Methotrexate (MTX), a widely used chemotherapeutic agent, often induces hepatotoxicity, limiting its clinical utility. Cannabidiol (CBD), derived from hemp, possesses antioxidant, anti-inflammatory, and antiapoptotic properties. This study aims to investigate CBD’s protective effects against MTX-induced liver injury and elucidate the underlying mechanisms. Thirty-two female Wistar Albino rats were divided into four groups: control, MTX (20 mg/kg intraperitoneally [i.p.] once), MTX+CBD (20 mg/kg i.p. once + 5 mg/kg i.p. for seven days), and CBD (5 mg/kg, i.p. for seven days). Biochemical analyses of serum and liver tissues were performed to assess oxidative stress markers (total oxidant status, total antioxidant status, oxidative stress index), liver function tests (AST, ALT), and antioxidant enzyme activities (glutathione peroxidase, superoxide dismutase). Histopathological and immunohistochemical examinations were conducted to evaluate liver tissue damage and TNF-α expression. Genetic analyses were performed to measure the expression levels of SIRT-1, p53, Bcl-2, and Bax genes using RT-qPCR. MTX administration increased oxidative stress markers, liver enzymes, TNF-α, p53, and Bax levels while decreasing antioxidant defenses and SIRT-1 expression. CBD administration reversed these alterations effectively. CBD mitigated MTX-induced hepatotoxicity by reducing oxidative stress, inflammation, and apoptosis. It activates antioxidant defenses via SIRT-1 upregulation, suppresses inflammation by reducing TNF-α, and prevents apoptosis by modulating p53, Bcl-2, and Bax gene expressions. These findings suggest CBD could be a promising therapeutic agent for chemotherapy-induced liver damage. Further research is warranted to explore additional pathways and broader molecular mechanisms.

Curcumin-Etoposide Synergy: Unveiling the Molecular Mechanisms of Enhanced Apoptosis and Chemoresistance Attenuation in Breast Cancer.

Combining natural compounds with chemotherapeutic agents has emerged as a promising approach for cancer treatment. Curcumin (Cur), a natural polyphenol, is known for its anti-cancer properties, including the ability to induce apoptosis and arrest cell cycle progression. This study aimed to evaluate the effects of Cur and etoposide (ETO), both individually and in combination, on the induction of apoptosis in breast cancer (BC) cell lines. The impact of Cur and ETO on cell proliferation was assessed using MTT viability assays. Apoptosis induction by these drugs was evaluated through Annexin V flow cytometry and caspase-3 and caspase-9 activity assays. Quantitative real-time PCR was employed to measure Bax and Bcl-2 gene expression levels. Western blotting was conducted to determine protein levels of p53, p21, Bax, and Bcl-2. A non-significant dose of ETO was selected based on MTT assay results and combined with 75 µM of Cur. Curcumin enhanced ETO's pro-apoptotic effect by increasing caspase activities. The combination of Cur and ETO significantly reduced Bcl-2 gene expression while upregulating Bax expression. Furthermore, treatment with this combination elevated the protein levels of p53, p21, and Bax, compared to ETO or Cur alone, while significantly decreasing Bcl-2 protein levels. Cur has the potential to amplify ETO-induced apoptosis in BC cells. This combination may offer a promising therapeutic approach for BC.

Electrospun nanofibrous scaffolds reinforced with therapeutic lithium/manganese-doped calcium phosphates: Advancing skin cancer therapy through apoptosis induction

In the current study we fabricated potent materials by incorporating therapeutic elements into calcium phosphates (CPs) to combat cancer. This involved synthesizing manganese (Mn)- and lithium (Li)-doped CPs and loading them into electrospun nanofibers (NFs) composed of chitosan (CS) and polyethylene oxide (PEO). The characterized CPs exhibited excellent properties, including a particle size of 47–75 nm, surface charge of –(30−56) mV, and specific surface area of 75–266 m2/g. The electrochemical analysis revealed that Mn and Mn/Li-doped CPs are promising for generating oxygen free radicals and H2O2, crucial for cancer therapy. Biological evaluation showcased the outstanding performance of the developed materials. MTT assay revealed a cytotoxic effect of nano-constructs on melanoma A375 cell line without adverse effects on normal L929 cells over 72 h. Annexin V/PI apoptosis assay indicated substantial apoptosis rates in A375 cells treated with PC-20 % (62.55 ± 4.59 %). The obtained data of qPCR analysis of pro-apoptotic and anti-apoptotic genes (P53, Bax, Bcl-2) in A375 cells treated with different CP nanoparticles (NPs) showed a significant increase in P53 and Bax gene expression, indicating high levels of A375 cell apoptosis. Additionally, the samples containing Mn ion exhibited high reactive oxygen species (ROS) generation. In conclusion, the fabricated NFs scaffolds hold promising potential for cancer therapy.

Morpho-molecular evaluation for developmental competence of oocytes retrieved through transvaginal ovum pick-up from FSH-stimulated Tharparkar donor cows (Bos indicus).

The study was conducted on indigenous Tharparkar cow (Bos indicus) to evaluate FSH stimulation on follicular attributes, oocyte recovery and morpho-molecular developmental competence parameters concerning oocyte quality. A total of 20 OPU sessions were performed, which included 10 sessions in each FSH stimulated at the dose of 130 µg divided into four sub-doses and non-stimulated. Findings on the size of follicles having ≥6 mm showed a significantly higher, however an opposite trend was observed in the case of smaller sized follicle (<6 mm) between stimulated and non-stimulated respectively. The stimulated cows had a significantly higher number as well as the percentage of oocytes of Grade A, having a diameter ≥120 µm and BCB+VE as compared to the non-stimulated cows. The relative mRNA expression profile of GDF9, BMP15, PCNA and BCL-2 genes was higher and BAX was lower in the FSH-stimulated cow. These results indicated that FSH stimulation before OPU in Bos indicus cows has a significant impact on follicle size, oocyte yield, recovery, and their quality with respect to COC's, diameter and BCB+VE oocytes. Further, a significant increase in the relative mRNA expression levels of GDF9, BMP15 and PCNA genes in the FSH-stimulated group suggests that FSH plays a key role in modulating the expression of these important candidate genes and thus influencing oocyte quality. The higher mRNA expression of BCL-2 genes and concomitantly lower expression of BAX gene in FSH Stimulated cows indicates the protective role of these genes and preventing programmed cell death and thus promoting cell survival, quality and embryo development.

Potential Protective Effect of Orlistat: A Formulation of Nanocrystals Targeting Inflammation, Oxidative Stress, and Apoptosis in an Experimental Model of Doxorubicin-Induced Cardiotoxicity.

Background: Doxorubicin (DOX) is a widely used chemotherapeutic agent; nevertheless, cardiotoxicity limits its effectiveness. Orlistat (Orli) is an irreversible lipase enzyme inhibitor with poor solubility and bioavailability. Furthermore, Orli has a favorable impact on the decrease in cardiometabolic risk variables. Thus, this study aimed to investigate the novel use of Orlistat Nanocrystals (Orli-Nanocrystals) to mitigate DOX-induced cardiotoxicity and to identify probable pathways behind the cardioprotective effects. Methods: The pharmacokinetic parameters-area under % dose/g heart time curve (AUC0→4h), Drug targeting index (DTI), and relative targeting efficiency (RTE)-were calculated. Furthermore, experimental design mice were categorized into six groups: a (1) Normal control group, (2) Orli-Free group, (3) Orli-Nanocrystals group, (4) DOX group, (5) Orli-Free-DOX group, and (6) Orli-Nanocrystals-DOX group. All treatments were intraperitoneally injected once daily for 14 days with a single dose of DOX (15 mg/kg) on the 12th day for 4, 5, and 6 groups. Results: The pharmacokinetic parameters (Cmax, AUC) following oral administration of Orli-Nanocrystals presented a significant difference (higher values) in comparison to Orli due to the enhanced extent of the absorption of nanocrystals and, subsequently, their distribution to the heart. The study results indicated that DOX caused significant cardiotoxicity, as revealed by a remarkable rise in cardiac function biomarkers like LDH and CK-MB, which involve enzyme activities. Additionally, cardiac MDA content also increased; however, glutathione peroxidase, catalase, and superoxide dismutase activities were decreased. In the same context, DOX was found to have a remarkable downregulation in Nrf2, HO-1, Sirt-1, and Bcl2, while the upregulation of NF-κB, TNF-α, and BAX gene and protein expression occurred. Pretreatment with Orli-Nanocrystals displayed the most notable recovery of the altered immunohistochemical, histological, and biochemical characteristics as compared to the Orli-Free group. Conclusions: This work is the first investigation into the potential use of antioxidant, anti-inflammatory, and anti-apoptotic characteristics of Orli-Nanocrystals to protect against DOX-induced cardiotoxicity in vivo.

Dendrosomal Curcumin Showed Cytotoxic Effects on Breast Cancer Cell Line by Inducing Mitochondrial Apoptosis Pathway and Cell Division Arrest.

Mutations in the p53 gene have been linked to the initiation and progression of breast cancer, as well as resistance to chemotherapy. Therefore, the development of novel treatment approaches is essential to combat this disease. This study aimed to evaluate the effects of dendrosomal curcumin (DNC) on the breast cancer cell line MDA-MB231. MDA-MB231 cells were treated with 20 μM DNC, and the apoptosis rate and cell proliferation cycles were assessed using flow cytometry. Additionally, after RNA extraction and cDNA synthesis, the expression levels of Lnc-DANCR, EZH2, Noxa, bcl-2, bax, PUMA, p21, and p53 genes were analyzed using RT-PCR. Protein expression levels of P53, P21, Bcl-2, and Bax were evaluated through western blotting. Dendrosomal curcumin induced apoptosis in MDA-MB231 cells and caused cell cycle arrest at the SubG1 phase. Dendrosomal curcumin treatment downregulated Lnc-DANCR, EZH2, bcl-2, and p53 gene expression, while upregulating bax, Noxa, PUMA, and p21 gene expression in a time-dependent manner. Bax and P21 protein levels were significantly upregulated following DNC treatment, whereas Bcl-2 and P53 protein levels were downregulated in DNC-treated breast cancer cells. In summary, dendrosomal nanocurcumin demonstrated potent anti-tumor effects against breast cancer cells, suggesting its potential as a therapeutic agent in breast cancer treatment.

The effect of L-carnitine in reactive oxygen species reduction and apoptotic gene expression in mice after cyclophosphamide: An experimental study.

Cyclophosphamide (CP), a utilized anticancer drug, is known to cause infertility in women. However, L-carnitine (LC), an antioxidant, has been shown to offer protective benefits against infertility. This study aimed to evaluate the levels of reactive oxygen species (ROS) and apoptotic gene expression in mice treated with CP and LC. 24 NMRI female mice (6-8 wk, 30 5 gr) were divided into 4 groups: control group: received normal saline intraperitoneal (IP) injection for 10 days; CP group: received 75 mg/kg of CP as a single IP on the 10 day of the experiment; LC group: received 200 mg/kg of LC IP for 10 days; LC+CP group: received LC for 10 days and CP single IP injection on the 10 day of the experiment. After 10 days, mice were superovulated. The oviducts were then removed, and the oocytes of each group were collected for evaluating apoptotic gene expression B-cell lymphoma 2(Bcl2), Bcl2-associated X(Bax), and Caspase3 via real-time polymerase chain reaction and intracellular ROS levels by dichloro-dihydro-fluorescein diacetate fluorescence staining. Data revealed that LC in the LC+CP group significantly increased Bcl2 gene expression (p = 0.01), and decreased Bax and Caspase3 gene expression compared to the CP group (p = 0.03, p = 0.04). LC decreased the ROS level in the LC+CP group compared to the CP group (p 0.001). Findings suggest that LC can scavenge the ROS caused by CP and modulate the apoptotic pathway via downregulating the Bax and Caspase3 genes and upregulating the Bcl2 gene in oocytes of mice exposed to CP.

Evaluation of antibacterial and anticancer properties of secondary metabolites isolated from soil Bacillus spp focusing on two strains of Bacillus licheniformis and Bacillus siamensis

BackgroundBacillus strains are well recognized for their inherent production of bioactive compounds that exhibit antibacterial and anticancer properties. This study aims to evaluate the antimicrobial and anticancer effects of the secondary metabolite isolated from Bacillus licheniformis and Bacillus siamensis strain.Material and methodWe developed and purified a new soil-derived Bacillus strain to study its metabolites on cancer cells and bacteria. After evaluating the antimicrobial effects of the selected strains’ secondary metabolites by well diffusion, growth conditions and temperature optimised using liquid-liquid extraction, secondary metabolites isolated, and active compounds identified using GC-MS. Evaluation of PC-3 and HPrEpC cytotoxicity. AV/PI staining and comet assay assessed necrosis and apoptosis. Real-time PCR measured apoptotic gene expression. Finally, the scratch test measured cell movement.ResultsBacillus strain metabolites exhibit dual-purpose antimicrobial and anticancer properties. Bacillus licheniformis isolate 56 and S2-G12 isolate 60 demonstrated the greatest antibacterial activity. Among all Bacillus isolates, isolates 56 (Bacillus licheniformis) and 60 (Bacillus siamensis strain) had the highest antibacterial activity. Crude extracts obtained from strains 56 and 60 decreased PC-3 cell viability in a dose-dependent manner. At 200 µg/mL, the survival rate of cells treated with strain 56 and 60 crude extract was 23% and 25%, respectively (p < 0.001). The treatment of PC-3 cells with strains 56 and 60 crude extract led to considerable apoptosis (46.2% and 50.09%, respectively) compared to the control group. After treatment with the crude extract from strains 56 and 60 at an IC50 concentration, a significant number of PC-3 cells showed comet formation, indicating DNA fragmentation. Metabolites extracted from strain 56 and 60 enhanced caspase 3, caspase 8, and Bax genes expression and reduced Bcl-2 expression (p < 0.001). Cell migration was also prevented.ConclusionOur findings show that the secondary metabolites of B. licheniformis and B. siamensis have antibiotic and anticancer properties. However in vivo studies are necessary to confirm these findings.

Exploring the effects of naringenin on cell functioning and energy synthesis in the hippocampus of male Wistar rats with chronic tinnitus, by examining genetic indicators such as Bax, Bcl-2, Tfam, and Pgc-1α

BackgroundThe pivotal factors, including neural plasticity, oxidative stress, neuronal inflammation, and apoptosis, play a significant role in the pathogenesis of tinnitus. The balance between Bax/Bcl-2 genes is an important factor in determining the rate of apoptosis. Pgc-1α and Tfam genes are fundamental regulators of mitochondrial biogenesis. Naringenin possesses significant antioxidant, neuroprotective, anti-inflammatory, anti-apoptotic, and antiviral properties, and its compounds are effective on cell signaling pathways. AimsIn light of the aforementioned information, we endeavored to evaluate the impact of naringenin on the expression levels of Bax, Bcl-2, Pgc-1α, and Tfam genes in the hippocampus of male Wistar rats with chronic tinnitus. Material and MethodsTo demonstrate the existence of tinnitus, all rats were instructed to complete an “active avoidance test” utilizing a conditioning box. The expression levels of genes mentioned above were assessed using real-time PCR. ResultsThe sodium salicylate at a dosage of 350 mg/kg showed an upregulation in the expression level of Bax and a downregulation in the expression level of the Bcl-2 gene (p < 0.001). Furthermore, the sodium salicylate displayed significantly higher expression levels of Tfam and Pgc-1α (p < 0.001) genes. The naringenin, at a dose of 100 mg/kg, led to a decrease in Bax gene expression (p < 0.05) and an increase in Bcl-2 gene expression (p < 0.05). On the other hand, naringenin restored the expression level of both Tfam (p < 0.001) and Pgc-1α (p < 0.01) genes. ConclusionsOur research findings demonstrate that sodium salicylate-induced tinnitus leads to enhanced apoptosis and mitochondrial biogenesis within the hippocampus. Additionally, our evidence recommends that naringenin can reduce apoptosis effectively and maintain a balanced mitochondrial state.