Bax Gene Expression Research Articles

Protective Effect of Spirulina against Aspartame Induced Oxidative Stress and Molecular Gene Brain damage

Spirulina and lipoic acid has been considered as one of the most effective in ant oxidative stress and anti-inflammatory activity.Aspartame (ASP) is one of the mostwidely used (artificial sweeteners)used in a variety of foods and feeds.This study was conducted to evaluate the possible protective effect of Spirulina and alpha-lipoic acid against aspartame induced oxidative stress and brain damage in rabbits. Forty two white male New-Zealand Rabbits were classified into seven equal groups. Group I: (Control group) received no drugs. Group II: rabbits administered with aspartame (250 mg/kg b. wt/day). Group III: rabbits received alpha- lipoic acid (100 mg/kg b. wt/day). Group IV: rabbits received spirulina platensis (1500 mg/kg. b. wt/day). Group V: rabbits received aspartame (250 mg mg/kg b. wt) and treated with alpha- lipoic acid (100 mg/kg b. wt). Group VI: rabbits received aspartame (250 mg mg/kg b.wt) and treated with spirulina platensis (1500 mg/kg b. wt). Group VII: rabbits received aspartame (250 mg mg/kg b.wt) and treated daily with alpha- lipoic acid (100 mg/kg b. wt) and spirulina platensis (1500 mg/kg b. wt) for 8 weeks. At the end of experiment brain tissue were isolated and analyzed for determination of L-malondialdehyde (L-MDA), Catalase (CAT) and reduced glutathione (GSH) in addition to Anti-inflammatory cytokines: Interlukin-10 (IL-10),Activator Protein-1(AP-1),Bax gene expression and DNA damage. The obtained results showed a significant up-regulation of Activator Protein-1(AP-1),Bax gene expression level and a significant down-regulation of Interlukin-10 (IL-10) and marked increase in L-MDA and DNA damage that was indicated by an increase in tail length and tail DNA percent in brain tissue of aspartame treated rabbits. However, brain CAT activity andGSH concentration were markedly decreased when compared with control group.Coad ministration of spirulinaand alpha lipoic acid protect aspartame induced brain damage in rabbits caused a significant improvement of all previous parameters and attenuate DNA changes. Conclusively, spirulina platensis and alpha-lipoic acid exerts a protective effect against DNA damage and oxidative stress in brain of aspartameusage through free radical scavenging and anti-inflammatory activities as well as regenerating endogenous antioxidants defense system mechanisms.

Exploring the Therapeutic Potentials of Exopolysaccharides Derived From Lactic Acid Bacteria and Bifidobacteria: Antioxidant, Antitumor, and Periodontal Regeneration.

The metabolites of lactic acid bacteria (LAB) and bifidobacteria (Bb) have recently received a lot of attention due to their ability to protect interactions in blood and tissues, as well as their biodegradability and biocompatibility in human tissue. Exopolysaccharides (EPS) derived from bacteria have a long history of use in therapeutic and other industrial applications with no adverse effects. In this regard, EPSs were isolated and characterized from LAB and Bb culture supernatants to determine their antioxidant, antitumor, and periodontal regeneration properties. The antioxidant capacity of the EPSs varied with concentration (0.625–20 mg/ml). The highest antioxidant activity was found in LAB: Streptococcus thermophiles DSM 24731-EPS1, Lactobacillus delbrueckii ssp. bulgaricus DSM 20081T-EPS5, Limosilactobacillus fermentum DSM 20049-EPS6, and Bb; Bifidobacterium longum ssp. longum DSM 200707-EPS10. Human breast cancer cells (MCF7), human colon cancer cells (CaCo2), human liver cancer cells (HepG2), and human embryonic kidney 293 (HEK 293) cells were used as controls to assess the antitumor properties of the selected EPSs. According to the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium-bromide (MTT) assay, EPS5 had the highest cytotoxicity against MCF7, CaCo2, and HepG2, with IC50 values of 7.91, 10.69, and 9.12 mg/ml, respectively. Lactate dehydrogenase (LDH) activity was significantly higher in cell lines treated with EPS5-IC50 values compared to other EPSs-IC50 values (p < 0.05). Real time (RT)-PCR results showed that EPS5 treatment increased Bax, Caspase 8, Caspase 3, and p53 gene expression. The expression of the BCL2, MCL1, and Vimentin genes, on the other hand, was reduced. The MTT test was used to examine the effect of EPS5 on the viability of human periodontal ligament fibroblast cells (hPDLFCs), and it was discovered that EPS5 increased hPDLFC viability. According to high-performance liquid chromatography (HPLC) analysis, galactose made up 12.5% of EPS5. The findings of this study pave the way for the use of EPS, which hold great promise for a variety of therapeutic purposes such as antioxidant, antitumor, and periodontal regeneration.

Ruthana date extract inhibited proliferation of human hepatocellular carcinoma (HepG2) cells by modulation of BAX gene.

Cancer response to chemotherapeutic agents and its side effects remain a challenge for the development of new anticancer compounds. Dates are consumed worldwide due to their high nutritional value. We investigated the cytotoxicity and expression of the proapoptotic BAX gene in human hepatocellular carcinoma (HepG2) cells treated with Ruthana date ethanolic extract (RDE). The RDE ingredients analyzed by GC/MS and HepG2 cells were treated with different concentrations of RDE for 24, 48, and 72h. Cytotoxicity, cell viability, DNA fragmentation, and BAX expression were determined. The GC/MS analysis of RDE showed its high content of quercetin, myricetin kaempferol, thymine, and catechol as the most active ingredients. HepG2 treated with RDE showed a significant change in morphological characteristics related to cell death. The antiproliferative activity determined by WST-1 demonstrated that RDE significantly reduced cell viability. Cells treated with RDE (10-60mg) showed gradual DNA fragmentation in a dose-dependent manner. Gene expression analysis showed upregulation of BAX at 30mg/ml of RDE (p < 0.001). However, it showed downregulation at (40-60mg/ml) as compared to control. Our findings indicated that RDE exert cytotoxicity against HepG2 cells due to its high content of flavonoids. This effect through DNA fragmentation and activation of the proapoptotic BAX gene.

Apoptotic effects of human amniotic fluid mesenchymal stem cells conditioned medium on human MCF-7 breast cancer cell line.

Breast cancer, as the most common malignancy among women, is shown to have a high mortality rate and resistance to chemotherapy. Research has shown the possible inhibitory role of Mesenchymal stem cells in curing cancer. Thus, the present work used human amniotic fluid mesenchymal stem cell-conditioned medium (hAFMSCs-CM) as an apoptotic reagent on the human MCF-7 breast cancer cell line. Conditioned medium (CM) was prepared from hAFMSCs. After treating MCF-7 cells with CM, a number of analytical procedures (MTT, real-time PCR, western blot, and flow cytometry) were recruited to evaluate the cell viability, Bax and Bcl-2 gene expression, P53 protein expression, and apoptosis, respectively. Human fibroblast cells (Hu02) were used as the negative control. In addition, an integrated approach to meta-analysis was performed. The MCF-7 cells' viability was decreased significantly after 24 hours (P < 0.0001) and 72 hours (P < 0.05) of treatment. Compared with the control cells, Bax gene's mRNA expression increased and Bcl-2's mRNA expression decreased considerably after treating for 24 hours with 80% hAFMSCs-CM (P = 0.0012, P < 0.0001, respectively); an increasing pattern in P53 protein expression could also be observed. The flow cytometry analysis indicated apoptosis. Results from literature mining and the integrated meta-analysis showed that hAFMSCs-CM is able to activate a molecular network where Bcl2 downregulation stands in harmony with the upregulation of P53, EIF5A, DDB2, and Bax, leading to the activation of apoptosis. Our finding demonstrated that hAFMSCs-CM presents apoptotic effect on MCF-7 cells; therefore, the application of hAFMSCs-CM, as a therapeutic reagent, can suppress breast cancer cells' viabilities and induce apoptosis.

Involvement of NF-κB/PI3K/AKT signaling pathway in the protective effect of prunetin against a diethylnitrosamine induced hepatocellular carcinogenesis in rats.

Prunetin (PRU) is an O-methylated flavonoid that is present in various natural plants and a primary significant compound found in isoflavone. Liver cancer creates major carcinogenic death despite recently advanced therapies. Hepatocellular carcinoma (HCC) treatment and prognosis are better in people with secure liver function. In the present study, we evaluated the action of PRU on diethylnitrosamine (DEN) alone HCC in a rat model through inflammation-mediated cell proliferative phosphoinositide 3-kinase/protein kinase B (PI3K/AKT) pathway analysis. Male Wistar rats were dividedinto four groups of six rats each. Group I, normal rats; Group II, DEN alone; Group III, DEN + PRU, and Group IV, PRU-alone. All groups of rats carried outhepatic cancer development by hypothesis antioxidant, biochemical, cell proliferative, apoptosis, cytokines protein, and gene expression status profiles. In tumor incidence DEN + PRU, 100% delayed the tumor growth disappearance of the lesion, and reversal of normal liver architecture was observed. Liver marker enzymes levels decreased when antioxidant levels (superoxidase dismutase, catalase, glutathione peroxidase, and glutathione reductase) were in Group III. Proinflammatory markers nuclear factor-κB, interleukin (IL)-6, IL-1β, and tumor necrosis factor α, were elevated in the rat's serum in Group III. Cell proliferative markers proliferating cell nuclear antigen and Cyclin-D1 protein expressions were downregulated; in contrast, Bcl-2, Bax, caspase-3, and caspase-9 gene expressions were upregulated and then it followed that protein expression of PI3K/AKT wasdownregulated in PRU-treated groups. PRU assisted reversal of liver damage, antioxidant enzyme restoration cytokine balance, protein, and gene expression to control levels. Taken together, PRU improves functions of the liver, and as such prevents HCC. PRU can be used together with chemopreventives for HCC.

Effect of Synthetic Cholesterol (Synthechol®) Supplementation in an Egg Yolk-free Extender on Dog Sperm Cryopreservation

BACKGROUND: SyntheChol® is a new synthetic, non-animal-derived cholesterol that is easily dissolved in ethanol, ready to use, and behaves in a similar way as natural cholesterol. Therefore, it could be used as a substitute of natural cholesterol in dog sperm freezing extender. OBJECTIVE: To evaluate the effect of supplementing an egg yolk-free (EY-free) extender with synthetic cholesterol (SyntheChol®) on cryopreserved dog sperm. MATERIALS AND METHODS: Spermatozoa (1 × 108 sperm/mL) were suspended in EY-free extender supplemented with 0% (control), 0.25, 0.5, 1, 2, 4, or 6% SyntheChol® (Extender 1), cooled at 4 °C for 1 h, and diluted (1:1, v/v) with Extender 1 containing 1 M glycerol. The spermatozoa were then cooled to 4 °C for 30 min. Sperm-containing straws were frozen using LN2 vapor. Sperm motility (computer-assisted sperm analysis, CASA), sperm membrane integrity (SYBR-14 and PI staining), and acrosome integrity (FITC-PSA) were evaluated after thawing. Thereafter, optimal concentrations were determined (0.25, 0.5, 1, or 2%) and used to evaluate reactive oxygen species (ROS) generation, apoptosis, and the gene expression of motility-related sperm mitochondria-associated cysteine-rich protein, apoptosis-related B-cell lymphoma 2 (BCL2), and BCL2-associated X protein ( BAX) in cryopreserved sperm. RESULTS: Sperm progressive motility, membrane integrity, and acrosome integrity were markedly greater in the SyntheChol®-supplemented groups (0.25, 0.5, 1, or 2%) than in the control group. Only BAX expression was significantly reduced in the SyntheChol® groups (0.25, 1, or 2%) compared with the control group. However, there were no significant effects on the ROS generation or apoptosis index. CONCLUSION: SyntheChol® (0.25, 1, or 2%) proved to be effective in reducing the BAX gene expression level and improving sperm progressive motility, and membrane and acrosome integrity.

The Anticancer Effect of a Conjugated Antimicrobial Peptide Against Colorectal Cancer (CRC) Cells.

Although antimicrobial peptides (AMPs) were initially known as compounds of the innate immune system to fight microbial pathogens, it has been recently proposed that differences in normal and cancer cell membranes cause the anticancer effect of these peptides. The aim of this study was to evaluate the anticancer effect of MELITININ+BMAP27-conjugated peptide against colorectal cancer (CRC) cells. The MELITININ+BMAP27-conjugated peptides were designed and the β-naphthylalanine residues were added to the termini to improve the anticancer effect. CRC cancer cell lines including HT29, SW742, HCT-116, and WiDr were used. After preparing concentrations of 5, 10, 25, 50, 100, 150, 200, and 400μg/mL of peptide solution, the rate of cell death after 12, 24, and 48h was assessed using MTT test. After confirmation of the 30µg/mL efficacy and nontoxic concentration, the cells were exposed to this concentration, and the total RNA was extracted. The quantitative real-time PCR (RT-qPCR) technique was performed for the amplification of Bax, caspase3, atg5, and GAPDH (glyceraldehyde 3-phosphate dehydrogenase as the internal control) genes. The cytotoxicity of peptide against normal cells exhibited that the IC50 at 24 and 4h included 80 and 100µg/mL, respectively. After 24-72h of treatment, a significant difference in the mean percentage of CRC living cells was observed at concentrations of 50-400μg/mL of conjugated peptide (p < 0.05). The IC50 of the peptide at 24, 48, and 72h of exposure was measured as 30, 20, and 10μg/mL, respectively. The peptide resulted in a significant increase of 2.35-fold in the mean expression of Bax gene in CRC cells (p < 0.001). It also caused a significant increase of 1.75 times (p = 0.0112) of caspase 3 gene and 1.2 times (p = 0.0217) of atg5 gene. There was no significant difference among cell lines regarding the expression of each gene. The conjugated peptide caused the death of CRC lines via induction of the apoptosis and necrosis mechanisms. More studies are needed in this regard.

Synergistic effect of quercetin and cobalt ferrite-graphene oxide-based hyperthermia to inhibit expression of heat shock proteins and induce apoptosis in breast cancer cells

Background: Combinatorial medicine includes promising therapeutic methods for diseases such as cancer, whereby various biochemical and physical agents are simultaneously used to remove tumors. For example, the effectiveness of hyperthermia as a new technique for cancer therapy, can be enhanced, if it is combined with chemical compounds. Herein, the influence of quercetin as a heat shock protein (HSP) inhibitor on the efficiency of cobalt ferrite-graphene oxide (CoFe2O4-GO) nanoparticles- based hyperthermia was investigated in an in vitro study. Methods: Firstly, the surface of graphene sheets was decorated with CoFe2O4 nanoparticles (5-8 nm) and assayed using transmission electron microscopy (TEM), vibrating-sample magnetometer (VSM) and X-ray diffraction (XRD) methods. The cytotoxic effect of the corresponding co-implementation was then examined in MCF7 cell line with or without hyperthermia by (3-(4,5-dimethyl thiazolyl-2)-2,5-diphenyltetrazolium bromide) (MTT) test for 24, 48 and 72 hrs. In addition, the expression of Bax, Bcl2 and HSP70 genes and the production of radicals were evaluated by Real-Time PCR and DPPH (2,2-diphenyl-1-picrylhydrazyl) assays respectively. Results: The study showed that the doses associated with the IC50 points for quercetin and the CoFe2O4-GO nanocomposite were 0.02 mg/ml and 0.001 g/ml, respectively. The results showed that the simultaneous treatment of the cancer cells with quercetin, the nanocomposite, and hyperthermia significantly improves the cytotoxicity effect, increases the expression of Bax gene and down-regulates HSP70 and Bcl2 genes. In addition, the greatest attenuation of DPPH free radicals was observed in the corresponding group. Conclusion: The hybrid treatment of quercetin and the nanoparticle in the presence of hyperthermia could be considered as a promising approach for cancer therapy with minor side effects.

Polydopamine Nanoparticles Loaded with EPB41L4A-AS2 Promote Ovarian Cancer Cell Apoptosis Through Activation of Mitogen-Activated Protein Kinase Signaling Pathway

Ovarian cancer (OC) is one of known gynecologic malignant tumors, posing a serious threat to women’s life, and this disorder is often not diagnosed until it’s more advanced. This study aimed to identify the efficacy of polydopamine (PDA) nanoparticles loaded with EPB41L4A-AS2 (EPB) on OC. PDA nanoparticles-loaded EPB (PDA-EPB) were prepared, and OC cells were administrated with 10 μmol/LPDA-EPB nanoparticles or 10 μmol LHBSS buffer (control group). Untreated cells were taken as control group, and CCK-8 assay was conducted 24 h after treatments, to detect cell proliferation, while Transwell assay was used to detect invasion. Apoptosis was evaluated by flow cytometry along with analysis of Bax, Bcl-2, and caspase-3 apoptosis genes expression as well as the expression of Erk1/2, JNK, and p38 proteins. PDA-EPB nanomaterials presented uniform nanoparticle size of 100 nm and hydrated particle size of 190 nm. Treatment with composite nanoparticles decreased proliferation of OC cells over time. The increased concentration of nanoparticles occurred with decreased proliferation activity and 10 μmol/L was the best intervention concentration. Besides, PDA-EPB nanomaterials resulted in decreased number of invaded and migrated cells (41.03±3.95%) and increased apoptosis rate (33.59±3.23%), compared with the other two groups. The apoptosis ability was significantly higher, P &lt; 0.05. Meanwhile, in the presence of PDA-EPB nanoparticles, the experimental group presented low expressions of Bad (1.96 ±0.23), Bcl-2 (1.56± 0.34), Caspase-3 (2.89± 0.28) and (0.98 ±0.39), JNK (0.53±0.31), and p38 (0.79±0.26). PDA-EPB nanoparticles therefore decreased cell proliferation and induced apoptosis through specifically binding to the MAPK signaling pathway to activate the expression of Erk1/2, JNK, and p38 in OC.

Leptin improves in-vitro maturation of goat oocytes through MAPK and JAK2/STAT3 pathways and affects gene expression of cumulus cells

We investigated whether the recombinant leptin (1, 10, 100 ng/mL) influences the meiotic maturation of goat oocytes, whether the MAPK and JAK2/STAT3 pathways mediate the effects of leptin during in-vitro maturation, and whether leptin differently affects the abundance of mRNAs relevant to leptin signal transduction and apoptosis in oocytes and cumulus cells. The addition of leptin to the maturation medium positively affected the number of oocytes that completed nuclear maturation. Nuclear oocyte maturation stimulated by leptin was significantly impaired when we added the specific inhibitors of MAPK (U0126) and JAK2/STAT3 (AG490) to the maturation medium. The addition of leptin (10 ng/mL) during maturation did not affect the expression of AMPKα1, PPARα, Caspase 3, and BCL2 genes in oocytes or cumulus cells. The PPARγ and BAX mRNA abundances were significantly reduced in cumulus cells in the leptin group compared to the control group. Our results demonstrate that supplementation of the in-vitro maturation medium with leptin significantly improves nuclear maturation and reveal the important role of the MAPK and JAK2/STAT3 signaling pathways in establishing the leptin-mediated nuclear maturation of goat oocytes. Moreover, leptin treatment affects PPARγ and BAX gene expression in cumulus cells.

The boosting effects of melatonin on the expression of related genes to oocyte maturation and antioxidant pathways: a polycystic ovary syndrome- mouse model

BackgroundMelatonin, as a free radical scavenger exhibiting genomic actions, regulates the antioxidant genes expression and apoptosis mechanisms. In polycystic ovary syndrome (PCOS) patients, an imbalance between free radicals and antioxidants in follicular fluid leads to oxidative stress, aberrant folliculogenesis, and intrinsic defects in PCOS oocytes. In this experimental mouse model study, oocytes of PCOS and the control groups were cultured in different melatonin concentrations (10− 5, 10− 6, and 10− 7 M) to investigate the expression of oocyte maturation-related genes (Gdf9/Bmp15), antioxidant-related genes (Gpx1/Sod1), apoptotic biomarkers (Bcl2/Bax) and total intracellular ROS levels.ResultsGdf9 and Bmp15, Gpx1 and Sod1 were up-regulated in PCOS and control oocytes cultured in all melatonin concentrations compared to those cultured in IVM basal medium (P < 0.05). A significant decrease in the total ROS level was observed in all groups cultured in the supplemented cultures. Melatonin increased Bcl2 and decreased Bax gene expression in PCOS and control oocytes compared to non-treated oocytes.ConclusionsMelatonin increased antioxidant gene expression and regulated the apoptosis pathway, effectively reducing the adverse effects of culture conditions on PCOS oocytes. Furthermore, it influenced the expression of oocyte maturation-related genes in PCOS, providing valuable support during the IVM process.

Protective Effect of Minocycline on Bax and Bcl-2 Gene Expression, Histological Damages and Oxidative Stress Induced by Ovarian Torsion in Adult Rats.

BackgroundMinocycline is a widely used bacteriostatic antibiotic with various functions. The aim of this study was to investigate impact of apoptotic genes in ovary of the torsion/detorsion treated rat model by minocycline.Materials and MethodsThis experimental study was performed in 32 female Wistar rats classified in four groups, including: i. sham, ii. TD: torsion/detorsion group received normal saline, iii. TDM: torsion/detorsion group treated with 40 mg/kg Minocycline, and iv. MC: healthy group received 40 mg/kg Minocycline. After treatment period (7 days), histoplogical parameters, oxidative stress markers and hormone profile of serum as well as the expression of Bax and Bcl-2 genes were measured in the ovary of rats.ResultsLevels of superoxide dismutase (SOD), glutathione peroxidase (GPX) and estrogen were decreased in the TD group and significantly increased in the treated groups (P=0.001). Levels of malondialdehyde (MDA) and testosterone were increased in the TD group and decreased in the treated groups (P=0.001). Expression level of Bax was elevated in the TD group, while it was attenuated in the treated groups (P=0.001). Expression level of Bcl-2 was significantly increased in treated groups (P=0.001).ConclusionMinocycline can repair oxidative damage in ovarian tissue and regulate apoptotic-related gene expressions.

Curculigo orchioides polysaccharides extraction, characterization, and their protective effects against femoral head necrosis

The use of steroid drugs such as dexamethasone in long-term treatment poses a challenge called femoral head necrosis. To reduce the destructive effects of dexamethasone, the use of herbal suppressants such as Curculigo orchioides polysaccharides (COPs) is recommended despite the ambiguities in their chemical composition and the effects of each component on the anti-necrosis activity of the femoral head. In this study, after separation of COPs through hot water–ethanol precipitation method and purification through DEAE-Sepharose fast flow column, their properties were explained by FTIR, NMR, methylation and chromatography. Moreover, the COPs biocompatibility and its inhibitory effects against dexamethasone-induced cytotoxicity were evaluated by MTT assay and gene expression on human primary osteoblasts cells. The results exhibit that the 4756 Da molecular weight COPs is generally composed of Rha, Ara, Fru, Xyl, Man, Glc, Gal, Glu A and Gal A. Also, MTT results recognized high biocompatibility of COPs and suppressive effect on dexamethasone. Also, COPs significantly reduced dexamethasone-induced intracellular levels of apoptosis and ROS. Furthermore, the use of COPs has significantly increased ALP activity, collagen content and mineralization, which are very effective in differentiating HPOS cells to repair bone tissue. Meanwhile, the gene expression outcomes indicate an increase in Bcl-2 expression gene and decrease in BAX, Caspase-9 and Caspase-3 gene expression in the presence of COPs, which are important in controlling apoptotic activity and bone regeneration. Overall, this study revealed that COPs can lead to the treatment of femoral head necrosis.

Inflammation reduction potential of nanostructured lipid carriers encapsulated with rat's bone marrow cells' lysate.

Bone marrow-derived mesenchymal stromal cells (BMSCs) have been used for treating inflammatory disorders. Due to the large size of BMSCs compared to nanoparticles, BMSCs cannot be loaded into the nanoparticles. It is hypothesized that BMSCs lysate loading into the nanocarriers will effectively deliver cellular contents and regulatory elements of BMSCs at the injury site. This study aimed to investigate nanostructured lipid carriers (NLC) loading with BMSCs lysate through basic characterization and morphological analysis. Moreover, this study was mainly designed to investigate the role of NLC loaded BMSCs lysate in reducing inflammation via in-vitro and in-vivoassays. The in-vitro study involves cell viability assays, p53, annexin V and VEGF expression through ELISA and immunocytochemistry, real-time BAX, caspase-3, IL-6, IL-8, TOP2A, PCNA, and Ki-67 gene expression analysis. Additionally, to evaluate in-vivo anti-inflammatory activity, the carrageenan-induced rat paw oedema model was used. In-vitro results showed that NLC loaded BMSCs lysate increased cell viability, decreased apoptosis and pro-inflammatory genes expression and up-regulated angiogenesis and proliferation in H2O2 pre-stimulated cells. Findings of the in-vivo assay also indicated a reduction in rat's paw oedema volume in NLC-loaded BMSCs lysate, and downregulation of BAX, Caspase-3, IL-6, and IL-8 was observed. Enhanced expressions of TOP2A, PCNA, and Ki-67 were obtained. Concluding the results of this study, NLC-loaded BMSCs lysate could reduce inflammation and possibly regenerate damaged tissue mainly via increasing cell viability, angiogenesis and proliferation, and reducing apoptosis and pro-inflammatory cytokines.

Coenzyme Q10 protects against doxorubicin-induced cardiomyopathy via antioxidant and anti-apoptotic pathway

ABSTRACT Doxorubicin (Dox) is an anthracycline antibiotic that treats a variety of malignancies. Unfortunately, its cardiotoxicity limits its therapeutic usefulness. Coenzyme Q10 (CoQ10) has effectively treated and prevented various cardiac diseases and toxicities. This study aimed to evaluate the possible antioxidative and anti-apoptotic cardioprotective effects of CoQ10 against doxorubicin-induced histopathological and molecular changes in cardiomyocytes. Twenty-eight adult Wistar rats were divided into positive control, negative control, Dox-treated group, and Dox+CoQ10-treated. On the 16th day after the start of treatment, the hearts of all rats were dissected, and the left ventricles were processed for histological evaluation; immunohistochemical staining with caspase-3 and inducible nitric oxide synthase (iNOS); ultrastructural examination of cardiomyocytes; molecular assessment of proapoptotic gene Bax and anti-apoptotic gene expression Bcl-2; and biochemical study of malondialdehyde (MDA). The Dox-treated group had disorganized cardiomyocytes with increased interstitial space, vacuolated cytoplasm, and multiple small-sized pyknotic nuclei. A significant increase in caspase-3 and iNOS immunoexpression was observed. Ultrastructurally, the mitochondria were large with abnormal shapes, vacuolated cytoplasm, multiple vacuoles and autophagosomes, collagen fibril accumulation, and multiple small hyperchromatic nuclei. The intercalated discs were disorganized with loss of desmosome junction. The cardiomyocytes also showed significantly increased MDA levels and upregulation of Bax/Bcl-2 gene expression ratio. Co-administration of CoQ10 resulted in significant improvement in the histopathological picture, with a significant decrease in caspase-3 and iNOS immunoexpression and downregulation of the Bax/Bcl-2 gene expression ratio. In conclusion, CoQ10 protects against Dox-induced cardiotoxicity through the regulation of proapoptotic and anti-apoptotic gene expression.

In vitro anticancer effects of ibuprofen on HeLa cell line

Several studies have reported the anticancer effects of ibuprofen on cervical cancer cells, but the molecular pathway is still unclear in many aspects. This study aimed to investigate the effects of cytotoxic dose of ibuprofen on BAX, BCL-2, caspase-3, MMP-9, KAI-1 and NM23 gene expression levels and evaluation of caspase-3, -8 and -9 activity level in cervical cancer (HeLa) cells. Cervical cancer cells were divided into untreated (control) groups and ibuprofen treated groups. Expression levels of BAX, BCL-2, caspase-3, MMP-9, KAI-1 and NM23 genes were evaluated by Real-time PCR and caspase-3, -8 and -9 activity levels were determined using colorimetric method. Hoechst staining was used to confirm apoptosis. The data were statistically analyzed between groups using ANOVA and independent t-test. Significant increase in expression levels of caspases-3, and BAX/BCL-2 ratio, caspase-3, -8 and -9 activity level and a significant decrease in NM23, KAI-1 genes expression level were observed in HeLa cells treated with ibuprofen cytotoxic concentration. The apoptosis observed in HeLa cells was confirmed by Hoechst 333285 staining and flow cytometry analysis. Ibuprofen increased nuclear condensation and induced apoptosis in HeLa cells by both intrinsic and extrinsic apoptosis pathways.

Apigenin Enhanced Radiation-Induced Apoptosis/Necrosis by Sensitization of LNCaP Prostate Cancer Cells to 6 MV Photon Beams.

Objective Whereas prostate cancer (PrCa) may be unresponsive or moderately responsive to radiation therapy (RT)- most common modality for treatment of PrCa- patients must receive a high dose of RT In order to achieve appropriate tumour control. However, this increase in radiation dose may lead to severe adverse effects in normal tissues. Sensitization of PrCa to radiation provides an alternate approach to improve the therapeutic efficacy of RT. This study aims to assess the radiosensitisation effect of apigenin (Api) on a prostate cancer cell line (LNCaP). Materials and Methods In this experimental study, LNCaP cells were treated with 0-80 µM Api to investigate its effect on LNCaP cell viability and determine its half-maximal inhibitory concentration (IC50). Next, the cells were divided into four groups: i. Control, ii. Cells treated with the IC50 concentration of Api, iii. Cells treated with 2 Gy ionizing radiation (IR), and cells co-treated with Api and IR. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, real-time polymerase chain reaction (PCR), and an Annexin V-FITC/PI assay were performed to assess cell survival, Bax and Bcl-2 expressions, and presence of apoptosis and necrosis. Results Api inhibited cell survival in a dose-dependent, but not time-dependent manner. Cells treated with Api had increased amounts of early apoptosis, late apoptosis, and secondary necrosis compared to the control group. This group also had decreased Bcl-2 gene expression and up-regulated Bax gene expression. Co-treatment with Api and IR significantly inhibited cell survival, and increased early apoptosis, late apoptosis and secondary necrosis compared to the other groups. There was a significant decrease in Bcl-2 gene expression along with up-regulation of Bax gene expression, and Bax/Bcl-2 ratio changes that favoured apoptosis. ConclusionApi inhibited PrCa cell survival and induced apoptosis as a single agent. In addition, Api significantly sensitized the LNCaP cells to IR and enhanced radiation-induced apoptosis.

Antineoplastic effectiveness of silver nanoparticles synthesized from Onopordum acanthium L. extract (AgNPs-OAL) toward MDA-MB231 breast cancer cells

The present research was done to investigate the anticancer properties of silver nanoparticles (AgNPs) fabricated using bioactive extract of Onopordum acanthium L. (AgNPs-OAL) against breast cancer cells MDA_MB231 in vitro. The determination studies of AgNPs-OAL were confirmed by X-ray diffraction (XRD), field emission scanning electron microscopy (FESEM) analysis. Interestingly, the FESEM image observed the spherical shape of AgNPs-OAL with the range of 1-100 nm. As AgNP-OAL exhibited significant cytotoxicity properties on breast cancer MDA_MB231 cells with IC50 values of 66.04µg/mL, while lowing toxicity toward normal human embryonic kidney 293 (HEK293) cells with IC50 values of 101.04µg/mL was evaluated. Further, up-regulation of apoptotic Bax and CAD gene expressions were confirmed by quantitative real-time reverse transcription-PCR (qRT-PCR) technique results. Moreover, enhanced cell cycle population (sub-G1), annexin V/PI staining, acridine orange and ethidium bromide (AO/EB) staining, Hoescht 33,258 dye, and generation of reactive oxygen species were observed in AgNP-OAL-treated MDA_MB231 cancer cells. The green-synthesized AgNP-OAL has promising anticancer efficiency that can trigger apoptosis pathways in the MDA_MB231 breast cancer cells.

Combined Effect of Cold Atmospheric Plasma and Curcumin in Melanoma Cancer.

Curcumin (CUR) has interesting properties to cure cancer. Cold atmospheric plasma (CAP) is also an emerging biomedical technique that has great potential for cancer treatment. Therefore, the combined effect of CAP and CUR on inducing cytotoxicity and apoptosis of melanoma cancer cells might be promising. Here, we investigated the combined effects of CAP and CUR on cytotoxicity and apoptosis in B16-F10 melanoma cancer cells compared to L929 normal cells using MTT method, acridine orange/ethidium bromide fluorescence microscopic assay, and Annexin V/PI flow cytometry. In addition, the activation of apoptosis pathways was evaluated using BCL2, BAX, and Caspase-3 (CASP3) gene expression and ratio of BAX to BCL2 (BAX/BCL2). Finally, in silico study was performed to suggest the molecular mechanism of this combination therapy on melanoma cancer. Results showed that although combination therapy with CUR and CAP has cytotoxic and apoptotic effects on cancer cells, it did not improve apoptosis rate in melanoma B16-F10 cancer cells compared to monotherapy with CAP or CUR. In addition, evaluation of gene expression in cancer cell line confirmed that CUR and CAP concomitant treatment did not enhance the expression of apoptotic genes. In silico analysis of docked model suggested that CUR blocks aquaporin- (AQP-) 1 channel and prevents penetration of CAP-induced ROS into the cells. In conclusion, combination therapy with CAP and CUR does not improve the anticancer effect of each alone.

TAL effector-dependent Bax gene expression in transgenic rice confers disease resistance to Xanthomonas oryzae pv. oryzae.

The hypersensitive response (HR) is a form of programmed cell death of plant cells occurring in the local region surrounding pathogen infection site to prevent the spread of infection by pathogens. Bax, a mammalian pro-apoptotic member of Bcl-2 family, triggers HR-like cell death when expressed in plants. However, constitutive expression of the Bax gene negatively affects plant growth and development. The Xa10 gene in rice (Oryza sativa) is an executor resistance (R) gene that confers race-specific disease resistance to Xanthomonas oryzae pv. oryzae strains harboring TAL effector gene AvrXa10. In this study, the Xa10 promoter was used to regulate heterologous expression of the Bax gene from mouse (Mus musculus) in Nicotiana benthamiana and rice. Cell death was induced in N. benthamiana after co-infiltration with the PXa10:Bax:TXa10 gene and the PPR1:AvrXa10:TNos gene. Transgenic rice plants carrying the PXa10:Bax:TXa10 gene conferred specific disease resistance to Xa10-incompatible X. oryzae pv. oryzae strain PXO99A(pHM1AvrXa10), but not to the Xa10-compatible strain PXO99A(pHM1). The resistance specificity was confirmed by the AvrXa10-dependent induction of the PXa10:Bax:TXa10 gene in transgenic rice. Our results demonstrated that the inducible expression of the Bax gene in transgenic rice was achieved through the control of the executor R gene promoter and the heterologous expression of the pro-apoptosis regulator gene in rice conferred disease resistance to X. oryzae pv. oryzae.