Bcl-2 Gene Expression Research Articles

Unraveling TGF-β1's Role in Mediating Fibrosis and Cell Death in Feline Kidney Cells.

Chronic kidney disease (CKD) is prevalent among older cats. The transforming growth factor beta 1 (TGF-β1) pathway is associated with renal fibrosis. TGF-β1 signaling through the non-canonical/smad-independent pathway activates mitogen-activated protein kinase (MAPK) signaling, which is linked to fibrosis and apoptosis. The MAPK pathway regulates the Bcl-2 protein family, which is known for its anti-apoptosis properties. This study aimed to quantify the mRNA expression of the TGFβ, MAPK, and Bcl2 genes and the protein expression of TGF-β1 and MAPK in feline kidney cells and tissue. A gene expression analysis was conducted using qPCR to calculate the relative gene expression, while the protein expression was assessed through Western blot analysis. Immunohistochemistry staining of TGF-β1 and MAPK was performed on feline kidney tissue. The results revealed the significant upregulation of TGFβ (p = 0.001) and considerable downregulation of Bcl2 (p = 0.010) in doxorubicin-treated feline kidney cells. The immunostaining levels of TGF-β1 and MAPK were higher in the kidney tissue of cats with CKD than in non-CKD cats. However, there was no difference in TGFβ, MAPK, or Bcl2 gene expression in CKD vs. non-CKD cats. The findings suggest that TGF-β1 and Bcl-2 are associated with renal fibrosis and apoptosis in feline kidney cells. A deeper understanding of the TGF-β1 pathway could enable veterinarians to monitor disease progression and mitigate complications in feline CKD.

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.

Vitamin D protects spermatogonia and Sertoli cells from heat stress damage by inhibiting NLRP3.

Cryptorchidism can damage cells in the cryptorchid testes due to elevated local temperatures, potentially impacting the fertility of the child in adulthood. Research indicates that vitamin D enhances sperm quality in adult males. This study aimed to explore whether vitamin D inhibits NLRP3 activation, thus helping to mitigate heat stress damage to testicular spermatogenic and Sertoli cells. Five cases of normal testicular tissue adjacent to a tumor (testis removed due to tumorous growth) and five cases of atrophied cryptorchid testicular tissue (testis removed) were analyzed for immunohistochemistry to determine NLRP3 expression in cryptorchid tissue. In Phase I, spermatogonia (GC-1) and Sertoli cells (TM4) were separated into blank and heat stress groups. Apoptosis, inflammatory factor levels, and the expression of Bcl-2 and NLRP3 genes and proteins were measured at 2, 6, and 10 h after heat stress treatment. In Phase II, the cells were re-cultured and divided into three groups: heat stress, siRNA + heat stress, and VD + heat stress. After 10 h, the apoptosis, inflammatory factor levels, and gene and protein expressions of Bcl-2 and NLRP3 were reassessed in each group. Immunohistochemistry indicated NLRP3 expression in cryptorchid tissue. Phase I, extending heat stress duration led to increased apoptosis in spermatogonia (GC-1) and testicular Sertoli cells (TM4), heightened levels of inflammatory factors, reduced BCL-2 expression, and elevated NLRP3 expression compared to the control group. Phase II, both the siRNA + heat stress and VD + heat stress groups showed decreased apoptosis in spermatogonia and Sertoli cells, lower inflammatory factor levels, increased BCL-2 expression, and decreased NLRP3 expression compared to the heat stress-only group, with statistically significant differences (P < 0.05). This is the first time we found the expression of NLRP3 in cryptorchidism. Vitamin D can inhibit the expression of NLRP3 and reduce the damage of heat stress on testicular spermatogenic cells and Sertoli cells, and play a protective role for testicular spermatogenic cells and Sertoli cells. This provides a theoretical basis for preserving testicular function during the "treatment gap" in boys with cryptorchidism who have not received surgical treatment.

Development of a Novel Peptide with RGD Tumor Homing Motif: Evaluation of its Anticancer Potential in Hepatocellular Carcinoma and Colon Cancer Cells

Background: Peptide-based therapy has emerged as a promising avenue for treating various disorders, and recent research has highlighted the potential of anti-cancer peptides (ACPs) in cancer treatment. In this context, this study aimed to design a novel peptide incorporating a tumor-homing peptide (RGD) and C-amidation to enhance its anticancer activity, particularly against liver (HepG2) and colon (HCT-116) cancer cell lines. Objectives: The primary objective was to design a peptide with improved anticancer properties by leveraging the tumor-homing capabilities of RGD and enhancing its activity through C-amidation. The study sought to evaluate the cytotoxicity of the designed peptide against red blood cells (RBCs) and normal Vero cells. Furthermore, the anticancer efficacy of the peptide was assessed in hepatocellular carcinoma (HepG2) and colon cancer (HCT-116) cell lines. The specific objectives included examining the apoptotic induction and morphological changes in treated cells compared to untreated cells. Methods: The peptide was designed using the ACPred-FL bioinformatics tool, and its cytotoxicity was assessed through hemolysis assays against RBCs and normal Vero cells. Anticancer activity was evaluated against HepG2 and HCT-116 cell lines. The analysis of apoptotic induction involved measuring the relative gene expression of oncogenic marker BCL2 and apoptotic markers (BAX, BID, CAS-8). Additionally, Cytopathological examination and Western Blot analysis were employed to study morphological changes and confirm the quantification of relevant markers. Results: The designed peptide, consisting of twelve amino acids with a molecular mass of 1230.6233 Da and an isoelectric point of 9.81, exhibited low erythrocyte lysis and minimal toxicity to normal cells. The IC50 values demonstrated significant anticancer activity against both HepG2 (36.49±2.6 μg/mL) and HCT-116 (11.03±2.5 μg/mL) cell lines. Treated cells exhibited a significant decrease in the oncogenic marker BCL2 and an upregulation of apoptotic markers (BAX, BID, CAS-8). Western Blot analysis confirmed these results in addition to cytopathological examination that scattered apoptotic and degenerative changes. Conclusion: The designed peptide is considered a patent product that displayed remarkable anticancer activity against hepatocellular carcinoma and colon cancer cell lines, effectively modulating apoptotic and oncogenic markers. These findings highlight the potential of the peptide as a therapeutic agent for cancer treatment, emphasizing its clinical significance in combating liver and colon cancers. Nonetheless, further research and development are warranted to explore the translational potential of this peptide in clinical studies.

Impact of cataranthine treatment on miRNA34 and miRNA29 levels in HepG2 cells and their association with the expression levels of Bcl-2 and Nrf2.

Cataranthine is an alkaloid used in the development of anti-cancer drugs. In this study, the effect of cataranthine is assessed by measuring the levels of miR-34 and miRNA-29, which are effective regulators of BCL-2 and NRF-2 gene expression, and their relation to the survival of HCC cells. This study used cataranthine, and the HepG2 cell line. The MTT test was used to determine the appropriate concentration of cataranthine for treatment (IC50). Oxidative stress status was assessed by evaluating TAC (total antioxidant capacity), TOS (total oxidant status), and MAD (malondialdehyde) levels. Flow cytometry was used to investigate apoptosis. The expression levels of Nrf2, Bcl2, miRNA34, and miRNA29 genes in HepG2 were evaluated by RT-PCR. We observed that cataranthine significantly reduced the levels of oxidative markers (MAD, and TOS) and, conversely, increased the level of antioxidant markers in HepG2 cells. Treatment of HepG2 cells with different doses of cataranthine significantly increased the expression of Nrf2 and Bcl-2 genes, while significantly decreasing the expression of miR29 and miR34 genes. These findings suggest that cataranthine may exert its anticancer effects by reducing oxidative stress and promoting apoptosis, while decrease in miR34 and miR29 as well as increase in Nrf2 and Bcl2 may act as resistance mechanisms in cancer cells. The results highlight the dual potential of cataranthine in regulating cellular responses to oxidative stress and cell death in liver cancer, with dose-dependent modulatory effects.

Tropisetron attenuates D-galactose-induced heart aging in male mice: activation of sirtuin1.

This study pursued to evaluate the tropisetron effects in attenuating D-galactose induced heart aging in mice. The study aimed to ascertain whether tropisetron affects apoptotic processes, mitochondrial oxidative stress, or inflammatory variables in cardiac tissue, presumably through the modulation of the SIRT1 signaling pathway or sirtuin 1. Aging was induced via administration of D-galactose (200 mg/kg, s.c.). Then, mice were treated with tropisetron (1, 3, and 5 mg/kg/day, i.p.). After 8 weeks, the key indicators of oxidative mitochondrial dysfunction, oxidative stress, pro-inflammatory cytokines, interleukin-6, tumor necrosis factor-α, and nitric oxide concentrations were evaluated. Additionally, the gene expressions of apoptotic regulators Bax and Bcl2, as well as SIRT1, were assessed using real-time PCR. Histological alterations and serum lactate dehydrogenase levels were also assessed. Tropisetron alleviated mitochondrial oxidative stress and inflammatory mediators while decreasing immune cell infiltration into cardiac tissue generated by D-galactose. The simultaneous injection of tropisetron effectively inhibited D-galactose-induced apoptosis by modulating the Bax/Bcl2 ratio and activating the SIRT1 pathway. The administration of tropisetron resulted in reduced serum lactate dehydrogenase levels compared to the group treated just with D-galactose. Moreover, tropisetron successfully reinstated mitochondrial activity and diminished D-galactose-induced aberrant nitric oxide generation. The research concludes that tropisetron may provide protection against cardiac aging by activating multiple mechanisms associated with the SIRT1 pathway.

Postbiotic metabolites derived from lactobacillus fermentum as potent antiproliferative bioresources on HeLa cells with promising biocompatibility

Chemotherapy administrations for cervical malignancy possess a variety of unfavorable influences on the human body. Scientists are interested in microbial-derived biomolecules or postbiotics as an alternative therapeutic strategy in malignant patients. This research investigated the mechanisms related to the function of two potential postbiotic Lactobacillus isolates, Lactobacillus fermentum CH and L. fermentum KH, isolated from indigenous Iranian dairy products. The Lactobacillus isolates were recognized through 16S rDNA sequence analysis followed by characterization using morphological and biochemical assays. The bioactivity of postbiotics on the cervical cancer model was also assessed through a cytotoxic study and apoptosis analysis. In addition, the anticancer activity was evaluated by qPCR, followed by a confirmation of the flow cytometry. The results of the bioactivity assay revealed that these postbiotics had suitable anticancer influences on the cervical cancer model (HeLa cells) by increasing BAX, caspase8, and caspase9, followed by a decrease in BCl-2, iKB (Inhibitor of nuclear factor kappa-B), and RelA gene expressions. Thus, the findings of this study signify that the postbiotic derivate from Lactobacillus strains isolated from indigenous Iranian dairy products could be regarded as a topical treatment with a promising curative index due to their effectiveness on cervical malignancy cells.

Novel therapeutic target for diabetic kidney disease through downregulation of miRNA-192-5p and miRNA-21-5p by celastrol: implication of autophagy, oxidative stress, and fibrosis.

One of the most common microvascular effects of diabetes mellitus (DM) that may result in end-stage renal failure is diabetic kidney disease (DKD). Current treatments carry a substantial residual risk of disease progression regardless of treatment. By modulating various molecular targets, pentacyclic triterpenoid celastrol has been found to possess curative properties in the treatment of diabetes and other inflammatory diseases. Therefore, the present study investigated whether celastrol has anti-inflammatory, antioxidant, and antifibrotic effects as a natural compound against experimental DKD. Streptozotocin (55mg/kg) was utilized for inducing DKD in a rat model. Antioxidant enzymes and renal function tests were assessed in serum samples. In kidney homogenate, relative miRNA-192-5p and miRNA-21-5p gene expressions were measured. Furthermore, using real-time PCR to evaluate the gene expressions of nucleus erythroid 2-related factor-2 (Nrf-2), matrix metalloproteinase-2 (MMP-2), proapoptotic caspase-3, antiapoptotic Bcl-2, LC-3, and Beclin-1. Moreover, the transforming growth factor β1 (TGF-β1), LC-3, Bcl-2,caspase-3and NADPH oxidase 4 (NOX4) renal expressions were assessed semi-quantitatively using immunohistochemistry. Seven weeks of celastrol (1.5mg/kg/day) treatment significantly ameliorated DKD. Celastrol improves kidney functions. Moreover, celastrol treatment demonstrated potent antioxidant effect. The mechanism of apoptosis resulting from the administration of celastrol included the modulation of Bcl-2 and caspase-3 expression in the kidney. Celasterol administration leads to an increase in LC-3 and Beclin-1 renal expression that resulting in autophagy. Celastrol treatment improved renal fibrosis by decreasing TGF-β1 and MMP-2 renal expression. These antifibrotic effects could be due to their ability to inhibit miRNA-192-5p and miRNA-21-5p expression in renal tissues. Celastrol exerts a renoprotective effect by targeting miRNA-21 and miRNA-192, as well as their downstream pathways, such as autophagy, apoptosis, and fibrosis.

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.

Tropisetron-Induced Apoptosis: A Study on SKOV3 Cell Viability and Gene Expression

Background: Ovarian tumors account for 1% of all childhood neoplasms and are the most common genital neoplasm in children. Considering the role of serotonin in the promotion of tumor growth and metastasis in some cancers, tropisetron as a 5-hydroxytryptamine (5-HT3) receptor antagonist can exert anti-neoplastic effects. The current study seeks to determine the association between the use of tropisetron and ovarian cancer­cell lines (SKOV3) by evaluating apoptotic regulators. Materials and Methods: In this experimental study, after the addition of tropisetron to SKOV3 cancer cells at the concentrations of 1, 10, and 100 μM, the MTT assay was employed to assess the cell viability percentage within 24, 48, and 72 hours. The level of expression of Bcl2 and Bax genes after 24 hours were determined by Real Time-PCR, and Caspase 3 enzyme activity was measured by the ELISA method. Differences between groups were statistically analyzed by one-way analysis of variance (ANOVA). A p-value less than 0.05 is considered to be statistically significant. Results: Based on the MTT test, tropisetron significantly decreased the viability of SKOV3 cancer cells. It decreased the levels of Bcl2 expression according to real-time PCR results (P =0.0015) but increased the expression levels of Bax (P = 0.011) in SKOV3 cells. Both caspase-3 enzyme activity and the ratio of Bax to Bcl2 expression (Bax/Bcl2) were increased (P = 0.008 and P = 0.011, respectively). Conclusion: Tropisetron could inhibit tumor cell growth via apoptosis induction and exert proapoptotic effects in an ovarian cancer ¬cell line. Therefore, it seems that it can be considered as a possible chemotherapy drug for ovarian cancer. However, more studies should be conducted in this regard.

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.

Screening of Lactic Acid Bacteria Strains from Chinese Fermented Food (Suanshui) and its Protective Effect on Acute Liver Injury in Mice.

Suanshui is a traditional fermented vegetable food in the southwest of China that is a rich source of probiotics. This study aimed to screen probiotics from Suanshui that have a protective effect against liver injury using in vivo and in vitro experiments. Eleven strains were isolated according to acid-producing capacity and probiotic properties, including antibacterial activity against pathogenic bacteria, tolerance to artificial gastrointestinal fluid and bile salts, heat resistance, antioxidant capacity, hydrophobicity, autoaggregation capacity, and adhesion capacity. Three probiotic strains, LAB36 (Lacticaseibacillus paracasei, (L. paracasei)), LAB19 (Lactiplantibacillus plantarum (L. plantarum)), and LAB51 (Limosilactobacillus fermentum (L. fermentum)), were obtained by the TOPSIS comprehensive evaluation. Furthermore, in an in vitro model of inflammation induced by LPS treatment of AML12, metabolites derived from the intestinal contents of mice treated with the three strains significantly decreased the gene expression of IL-1β, TNF-α, and Caspase 3 in the LAB (19, 36, 51) + LPS group (P < 0.05) and significantly increased the gene expression of Bcl-2 compared with those in the untreated LPS group (P < 0.05). In addition, the LAB36 + LPS group also exhibited significantly decreased gene expression of TLR4 and Bax (P < 0.05). In vivo experiments, LAB36 pretreatment significantly decreased the levels of ALT, AST, and inflammatory factors (IL-1β and IL-6) in the serum, improved the antioxidant capacity, and reduced apoptosis in the livers of the model mice (liver injury induced by D-GalN/LPS). In summary, LAB36 isolated from Suanshui has a protective effect on liver injury.

Fluorescent Staining-Assisted Evaluation of the Anticancer Potential of Nyctanthes arbor-tristis Extracts in HepG2 Liver Cancer Cells.

The current study's initial goal was to investigate the anticancer activity of the extracts from the flowering plant Nyctanthes arbor-tristis. The ethanol extracts' phytochemical analysis was qualitatively determined. It was determined that the various phytochemical substances had antibacterial, antioxidant, and anticancer effects. GC-MS analysis revealed compounds like maleic anhydride, butanedioic acid, and oxalic acid emphasize the antioxidant and anticancer properties. MTT assay was investigated against three different cell lines (HepG2, A549, and MCF7). High level of resistance to HepG2 liver cancer cell lines was noted with ethanol flower extracts with IC50 value of 67.98 μg/mL. EtBr staining assay revealed HepG2 apoptotic cells displaying nuclear and membrane alterations when exposed to 70 μg/mL of ethanol extract. The cells treated with 70 μg/mL of extract showed a reduction of mitochondrial membrane potential, as evidenced by the MMP assay, which showed red fluorescence decreasing and green fluorescence increasing. DNA fragmentation analysis showed HepG2 cancer cells showed no signs of DNA fragmentation after being treated with 50 μM treatments of flower extracts for 72 h. Quantitative RT-PCR analysis showed increased Bcl2 gene expression with fold variation level of about 0.086094579. The obtained results emphasized the significance of Nyctanthes arbor-tristis extracts with a suggestion of chemotherapeutic medication to treat various cancers in the future.

In vitro and in vivo evaluation of anti-tumorigenesis potential of nano silver for gastric cancer cells.

Silver nanoparticles (AgNP) exhibit significant cytotoxicity against MKN45 cells (IC50: 105.5µg/mL). In vivo, AgNP at 150mg/kg induces necrosis, reduces proliferation, and alters gene expression, presenting a promising gastric cancer treatment strategy. Gastric cancer is the second leading cause of death from cancer worldwide. In this study, the anticancer effect of silver nanoparticles (AgNP) was evaluated in both In vitro and In vivo. First, an MTT assay was employed to estimate the cytotoxicity of AgNP. Next, the obtained IC50s were used as the main doses that were administrated. Regarding In Vitro, MKN45 cells were applied to induce tumor, and AgNP was administrated to mice at doses of 75 and 150mg/kg for 28 days twice a week in treatment groups post-induction of cancer. After 28 days, the expressions of the BAX, BCL2, and CXCR1 genes were evaluated. An immunohistochemical examination of CD34 and Ki67 markers and tissue absorption of silver nanoparticles were also performed. Our MTT assay results showed that AgNP's IC50 after 8, 24, and 48h were 105.5, 70.8, and 22.4µg/mL, respectively. In addition, the mean survival probability in the treatment groups was more than 25 days. It seemed that the effectiveness of the concentration of 150mg/kg of silver nanoparticles had caused a significant amount of necrosis in the tumor cells. In addition, the proliferation rate was decreased significantly in the 150mg/kg group, and the expression of CD34 and Ki67 markers was reduced significantly. However, the expression of BAX and BCL2 genes was increased in the treatment groups. So, as it was shown in this research in both In vitro and In vivo aspects, it seems that the administration of silver nanoparticles can represent a promising strategy in the treatment of gastric cancer.

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.

Pharmacotherapeutic potential of pratensein to avert metribuzin instigated hepatotoxicity via regulating TGF-β1, PI3K/Akt, Nrf-2/Keap-1 and NF-κB pathway

Metribuzin (MBN) is a selective herbicide that adversely damages the vital organs of the body including the liver. Pratensein (PTN) is a novel flavonoid that exhibits marvelous medicinal properties. This experimental trial commenced to elucidate the pharmacotherapeutic strength of PTN to counteract MBN provoked liver toxicity in rats. Thirty-six male albino rats (Rattus norvegicus) were categorized into four groups i.e., the control, MBN (133.33 mg/kg), MBN (133.33 mg/kg) + PTN (20 mg/kg) and PTN (20 mg/kg) alone treated group. Our findings revealed that MBN exposure promoted the expressions of Keap-1 as well as concentrations of ROS and MDA while reducing the gene expressions of Nrf-2 as well as activities of catalase (CAT), glutathione Peroxidase (GPx), glutathione reductase (GSR), heme oxygenase-1 (HO-1), superoxide dismutase (SOD) and glutathione (GSH) contents. The levels of albumin and total proteins were reduced whereas the levels of alkaline phosphatase (ALP), gamma-glutamyl transferase (GGT), aspartate aminotransferase (AST) and alanine aminotransferase (ALT) were enhanced following the MBN administration. Moreover, the gene expression of transforming growth Factor–β1 (TGF-β1), matrix metallopeptidase-2 (MMP-2), matrix metallopeptidase-9 (MMP-9), collagen, type I, alpha 1 and type-3 alpha were escalated in response to MBN intoxication. Furthermore, MBN administration cause a sudden upregulation in the levels of NF-κB, IL-1β, TNF-α, IL-6 & COX-2. Besides, MBN exposure enhanced the gene expression of Bax and Caspase-3 while reducing the gene expression of PI3K, Akt and Bcl-2. Additionally, MBN exposure dysregulated the normal histology of liver. However, PTN treatment notably protected the hepatic tissues via regulating abovementioned dysregulations due to its marvelous ROS scavenging potential.

Effects of alpha-lipoic acid and sildenafil citrate on sperm quality in asthenozoospermic men during freezing-thawing processes

We aimed to investigate the combined effects of alpha-lipoic acid (ALA) and sildenafil citrate (SC) on sperm quality during freeze-thawing in asthenozoospermic men. We categorized thirty semen samples from asthenozoospermic into five different groups for analysis: Control (fresh), Freeze, Freeze + SC, Freeze + ALA, and Freeze + SC + ALA. We evaluated sperm parameters in each group, including motility, viability, morphology, plasma membrane integrity, DNA fragmentation, mitochondrial membrane potential, protamine deficiency, acrosome integrity, malondialdehyde, tumor necrosis factor-alpha (TNF-α), antioxidants (Superoxide dismutase, Glutathione, Catalase), and gene expression of HSP70 and Bcl-2. The Freeze group showed significant declines in viability, motility, morphology, membrane integrity, antioxidants, and mitochondrial membrane potential, with increases in protamine deficiency, abnormalities, DNA fragmentation, TNF-α, and malondialdehyde compared to control (p = 0.000). These effects were significantly reversed in all treatment groups (p = 0.000). Bcl-2 and HSP70 expression increased significantly in all groups (p = 0.000). Acrosomal integrity decreased significantly (p = 0.000) in the Freeze group compared to controls and in the Freeze + SC group compared to the Freeze group. However, acrosomal integrity significantly increased (p = 0.000) in the Freeze + SC + ALA group compared to the Freeze + SC group and in the Freeze + ALA group compared to other treatments. Our findings indicate that the simultaneous addition of SC and ALA in the freezing media yielded increased sperm quality. This enhancement was evidenced by reductions in oxidative stress, inflammation, and apoptosis, along with partial prevention of premature acrosome reactions induced by SC.

Regulation of Apoptosis, Autophagy, and Metastasis by Luteolin in Human Bladder Cancer EJ138 Cells: An Experimental Study

Background: Chemotherapy remains a primary approach to cancer treatment, widely applied in bladder cancer (BC). However, the various side effects and resistance associated with chemotherapeutic drugs pose significant challenges in BC therapy, prompting interest in natural compounds like luteolin. Studies focus on its effects on key biological processes involved in BC, including metastasis, apoptosis, and autophagy. Objectives: This study investigated the regulation of mRNA expression of genes associated with apoptosis (BCL2, P53), autophagy (ULK1, ATG12), and metastasis (MMP2, MMP9) in malignant BC cells treated with luteolin. Methods: This was an in vitro experimental study. EJ138 BC cells were treated with various concentrations of luteolin, and its impact on cell viability, proliferation, and gene expression was assessed. The cytotoxic effect of luteolin on EJ138 BC cells was evaluated using the MTT assay after 24- and 48-hour treatments with different luteolin concentrations. Flow cytometry was performed to examine luteolin’s anti-proliferative effect, and RT-PCR was used to analyze mRNA expression of BCL2, P53, ULK1, ATG12, MMP2, and MMP9 genes. Results: MTT assay results confirmed that luteolin reduced the proliferation rate of BC cells. Flow cytometry indicated increased cell death in EJ138 BC cells following luteolin treatment. RT-PCR findings demonstrated that luteolin upregulated P53, ULK1, and ATG12 expression while downregulating BCL2 mRNA expression. However, luteolin treatment in EJ138 cells did not significantly alter MMP2 and MMP9 expression levels. Conclusions: These findings indicate that luteolin exerts cytotoxic effects on EJ138 BC cells by dysregulating mRNA expression of genes involved in apoptosis and autophagy. Therefore, luteolin shows potential as an effective anti-cancer agent for BC therapy.

Curcumin Induces MCF-7 Breast Cancer Cell Apoptosis Through miR-15a Alteration

Background: In recent years, the application of herbal compounds in cancer treatment has shown significant progress. Curcumin (CUR), a natural polyphenol, demonstrates potent anti-cancer effects against various cancers, including breast cancer (BC). Curcumin targets a range of molecular pathways, contributing to the inhibition of cancer cell proliferation, metastasis, and angiogenesis, and promoting apoptosis. Objectives: This study aimed to assess the effect of CUR on BC cells, focusing on alterations in the expression levels of Mir-15a and Bcl-2 through the apoptotic pathway. Methods: The cytotoxicity of CUR was evaluated using an MTT assay. Changes in the expression of Mir-15a and Bcl-2 genes were analyzed by real-time PCR, and cell apoptosis was measured using flow cytometry. Data analysis was conducted using SPSS. Results: The MTT assay results indicated that cell viability decreased as CUR concentration increased (5 – 25 µM). Real-time PCR results showed a significant decrease in the expression of Mir-15a and Bcl-2 (P &lt; 0.05). Flow cytometry findings demonstrated that CUR treatment at the IC50 concentration for 48 hours induced apoptosis in 75.9% of MCF-7 cells. Conclusions: Our study provides a crucial foundation indicating that CUR induces apoptosis in MCF-7 cells by modulating the expression of Mir-15a.

Chitosan-encapsulated selenium nanoparticles alleviate CCl4 induced hepatotoxicity through synergistically modulating NF-κB and Nrf2 signaling pathways and regulating Bcl-2 and Caspase-3 expression: A comprehensive study with multiple regression analysis

BackgroundThe delivery of selenium in a nano-form (Se-NPs) is a promising modality of treatment for various oxidative stress-induced diseases. ObjectiveThis study aims to investigate the conceivable effects of selenium nanoparticles either alone (Se-NPs) or encapsulated with chitosan (Se-CS-NPs) on toxicity induced by CCl4 in rats. MethodsEighty albino rats were divided equally into eight groups. The first group was the placebo. The second group was a positive control, while the third and the fourth groups got orally (Se-NPs 5 mg/Kg) and (Se-CS-NPs 225 mg/Kg) respectively. The fifth and sixth groups were protective groups in which Se-NPs or Se-CS-NPs were given simultaneously. The seventh and eighth groups were therapeutic as they received either Se-NPs or Se-CS-NPs after stopping the CCl4 injection for 4 weeks more. ResultsOur results showed that the protective and therapeutic groups showed an increase in caspase-3 gene expression with a decline in the expression of Bcl-2, Nrf2, and AFP genes. Histopathological and immunohistochemical investigations showed the role of selenium nanoparticles either alone or coated with chitosan in decreasing fibrotic marker collagen I positive reaction ConclusionSelenium nanoparticles showed an excellent effect in counteracting the toxic effect of carbon tetrachloride on liver functions, inflammation reactions, and apoptosis process. Moreover, using selenium nanoparticles has a strong role in preserving the liver architecture with its normal constituents. No additional benefit was observed when the selenium nanoparticles were encapsulated with chitosan.