Bax Gene Expression Research Articles

Effect of cold plasma and elicitors on bioactive contents, antioxidant activity and cytotoxicity of Thai rat-tailed radish microgreens.

Raphanus sativus var. caudatus or Thai rat-tailed radish (RTR) contains glucosinolates and isothiocyanates with chemopreventive effects; however, only mature plants have been investigated to date. Thus, the present study aimed to determine isothiocyanates, phenolic compounds and flavonoid compounds, antioxidant activity, cytotoxicity, and antiproliferative activity of RTR microgreens grown from seeds treated with cold plasma (21 kV for 5 min), organic elicitor (160 mmol L-1 NaCl, 10 mmol L-1 CaCl2 or 176 mmol L-1 sucrose) or both in combination. Seeds were germinated on vermiculite and sprayed with deionized water or elicitor for 7 days before harvest. Cold plasma had insignificant effect on growth, whereas NaCl and CaCl2 increased fresh weight. Plasma with CaCl2 led to the highest total isothiocyanate (ITC) content [1.99 g kg-1 dry weight (DW)] in RTR microgreens containing raphasatin as the only ITC detected. Plasma treatment gave the highest total phenolic content (7.56 mg gallic acid equivalents g-1 DW), antioxidant activity from a 2,2-diphenyl-1-picrylhydrazyl assay (7.70 mg trolox equivalents g-1 DW) and ferric reducing antioxidant power assay (21.72 mg Fe2+ g-1 DW). Microgreen extracts from plasma showed an IC50 value of 29.28 and 13.83 μg mL-1 towards MCF-7 and HepG2, respectively, with inhibitory properties on matrix metalloproteinase (MMP)-2 and MMP-9 proteins. Plasma enhanced Bax and Caspase-3 gene expression but reduced Bcl-2 and MMP-9 expression, indicating activation of apoptosis. Cold plasma shows promise as an innovative tool to enhance bioactive compounds with chemopreventive benefits in microgreens. © 2020 Society of Chemical Industry.

The effects of melatonin and vitamin D3 on the gene expression of BCl-2 and BAX in MCF-7 breast cancer cell line

ObjectiveCompelling evidence from epidemiological and experimental studies indicate that the disruption of circadian rhythm and decreased melatonin synthesis are risk factors for breast cancer. With the ever-increasing rate of breast cancer cases globally, there is a strong need for safer, effective, and alternate treatment strategies with lesser side effects. This study aimed to investigate the effects of melatonin and vitamin D3 on the gene expression of two apoptotic factors, BCL-2 and Bax, in the MCF-7 cancer cell line. MethodsCell viability (MCF-7) was studied through MTT reduction assay in response to melatonin (1 nM, 5 nM and 10 nM) and vitamin D3 (0.5 nM, 1 nM and 10 NM). The optimum dose of melatonin (5 nM) and vitamin D3 (0.5 nM) were selected for their combination and Bax/BCL-2 ratio determination. Changes on the expression of genes BCL-2 and Bax, were determined through mRNA expression while Western blot analysis was used to detect changes in the expression of BCL-2 and Bax at the protein level. ResultsThere was an upregulation of Bax gene expression and protein expression with the treatments of melatonin, vitamin D3, and their combination, with concomitant downregulation of Bcl-2 gene and protein expression. The Bax/BCL-2 ratio was increased significantly (p < 0.001) suggesting potential therapeutic treatments. ConclusionThe results suggest that treatment with melatonin and vitamin D3 inhibits the proliferation and induced apoptosis in breast cancer cells. We report here for the first time that this combination has effectively activated Bax and downregulated BCL-2 at the DNA and protein level.

Antineoplastic activity of Salmonella Typhimurium outer membrane nanovesicles

Nano-sized Gram-negative bacterial outer membrane vesicles possess unique structural and immunostimulatory effects that could be exploited to regress tumors by alerting the host immune system and reversing the immunosuppressive tumor microenvironment. The current study was conducted to investigate the antitumor activity of the outer membrane vesicles (ST-OMVs) of Salmonella Typhimurium ATCC 14028, in vitro in human colorectal carcinoma (HTC116), breast cancer (MCF-7), and hepatocellular carcinoma (HepG2) cell lines and in vivo in Ehrlich solid carcinoma-bearing mice model either as a mono-immunotherapy or as an adjuvant to a commonly used conventional chemotherapy. In addition, we investigated the safety of ST-OMVs. Adult Swiss albino female mice with transplanted Ehrlich solid carcinoma were treated with either ST-OMVs, paclitaxel or a combination of both. Tumor volume, growth inhibition rate, quantitative RT-PCR of Bax and VEGF genes expression, histopathology and immune-expression of caspase-3, Beclin-1, CD49b and Ki-67 were all analyzed. Our results showed that ST-OMVs significantly decreased tumor volume, significantly increased tumor growth inhibition rate, up-regulated the immunohistochemical expression of caspase-3, Beclin-1, and CD49b (enhanced recruitment of NK cells). Furthermore, ST-OMVs down-regulated the expression of Ki-67, increased Bax gene expression and decreased VEGF gene expression as detected by qRT-PCR analysis. Histologically, ST-OMVs promoted apoptosis, decreased tumor invasion and mitotic activities. Moreover, ST-OMVs showed a remarkable cytotoxic activity in various investigated in vitro cancer cell lines. Our findings demonstrate potential antitumor activity of ST-OMVs that might be used as a promising safe antitumor immunotherapy or an adjuvant to conventional chemotherapeutic drugs, resolving some of their problems.

Cytotoxic and Apoptotic Effects of Ferulic Acid on Renal Carcinoma Cell Line (ACHN)

Background: Polyphenolic compounds have anti-proliferative effects and can trigger apoptosis in cancer cells. Ferulic acid is excessively found in various herbal products and fruits. Ferulic acid plays a role in the treatment of neurodegenerative diseases, cancer, diabetes, cardiovascular disease, inflammation, and bacterial and viral infections. Objectives: The purpose of this study was to evaluate the anticancer effect of ferulic acid on renal carcinoma cells (ACHN). Methods: To assess the anti-proliferative effect of ferulic acid, the renal carcinoma cell line (ACHN) was treated with different ferulic acid concentrations (10, 20, 40, 80, and 160 μM) for 24, 48, and 72 hours. Cell viability was measured using the MTT assay. The apoptosis of cancer cells was evaluated by flow cytometry and real-time PCR. Results: The IC50 of ferulic acid against ACHN cells was determined to be 30 μM at 72 hours with the MTT assay. The treatment of cells with ferulic acid concentrations of 30 and 60 μM caused a significant increase in the apoptosis index. The Bcl-2 gene expression level was significantly lower in the treated group than in the control group, and the Bax gene expression level was significantly higher in the treated group than in the control group (P &lt; 0.001). Conclusions: The results of this study showed the apoptotic activity of ferulic acid against ACHN cells. These results can be helpful in the better understanding of the anticancer mechanism of ferulic acid, suggesting this substance as an alternative drug or in combination with conventional chemical treatments for cancer treatment.

Inhibitory Effects of Agaricus Blazei Murrill (FA-2-b-β) on Proliferation of Chronic Myeloid Leukemia Cells in vivo and Its Mechanism

To investigated the anti-tumor in vivo effect and mechanism of the acid RNA protein complex (FA-2-b-β) of Agaricus blazei Murrill extract. CCK-8 method was used to detected the inhibitory effect of FA-2-b-β on proliferation of primary CML cells from newly diagnosed CML patients, the CML mouse model was established by trail-venous injection of primary CML cells, and the survival time, blood cell count and body weight were observed, the immunoflouresence and immunehistochemistry analysis, RT-qPCR, Western bolt were used to detemine the expression of caspase-3 signal pathway-related apoptosis genes and proteins. The experiments in vitro showed that the proliferative inhibitory rate in drug-treated group increased with concentration- and time-dependent manner (r24=0.9092, r48=0.9442, r72=0.9546), the inter group comparison showed the statistical difference of results. The experiments in vitro showed that the survival time prolonged, blood cell count increased and body weight recovered in FA-2-b-β-treated group and imatinib-treated group, despite the WBC count is still high. The RT-qPCR and Western blot showed that the expression of BAX and caspase-3 gene and protein were up-regulated, the expression of BCL-2, cytochroime C, caspase-8, caspase-9 and BCL-ABL gene and protein were down-regulated. The FA-2-b-β can induce apoptosis of primary CML cells and prolong the survival time of CML model mouse, which may be related with the caspase-3 signal pathway related genes and proteins.

English

Avian influenza virus (AIV) A H5N1 is able to cause zoonosis with high morbidity and mortality. Several studies suggested the regulation of cellular apoptosis for virus survival by influenza viruses. This sheds light on the development of antivirals that can improve cellular apoptosis to fight influenza infections. This study aimed to investigate the regulation of pro-apoptotic gene expressions by H5N1 in MDCK cells. The H5N1 infection (102.67TCID50/ml) was performed for 8, 24, 48, 72, 96 and 120 hours. Upon incubation, the percentage of cell death, DNA fragmentation and pro-apoptotic gene expressions i.e. Bax, TRAIL, Caspase 3 (Cas3) and 8 (Cas8), and Fas-ligand (FasL) were determined. The degree of DNA laddering and pro-apoptotic gene expressions were determined on agarose gels. The results indicated that severe cytopathic effects (CPE) caused by H5N1 appeared as early as 24 hour post-infection (hpi) meanwhile a significant cell death was observed starting at 48 hpi. However, DNA laddering was not observed in both of the control and infected DNA samples. Downregulation of Bax gene expression was recorded at 24 -72 hpi and a total suppression was observed at 96 and 120 hpi. Similarly, a significant decrease in the TRAIL gene expression was observed at 48-120 hpi. On the contrary, the expressions of Cas3, Cas8 and FasL genes were not detected in the mock and infected samples. Conclusively, this study suggested that H5N1 might downregulate cellular apoptosis prematurely for viral survival which can be a target cellular mechanism for newly developed antivirals to combat H5N1 infections

Synergistic anticancer effects of phycocyanin and Citrullus colocynthis extract against WiDr, HCT-15 and HCT-116 colon cancer cell lines

BackgroundWe investigated the effect of C. colocynthis extract and phycocyanin on the growth of colorectal cancer (CRC) cells, and assessed the expression of caspase 3 and Bax encoding genes. MethodsWiDr, HCT-116 and HCT-15 CRC cells were treated with various concentrations of C. colocynthis and phycocyanin extracts for 24, 48 & 72 h. The cell cytotoxicity was measured by MTT assay. Expression of Caspase 3 and Bax genes was evaluated by quantitative real-time PCR (RT-qPCR). ResultsThe combination of extract + phycocyanin as well as C. colocynthis singly unraveled higher inhibitory effects against cancer cells compared to phycocyanin and control. The IC50 values for phycocyanin and the extract + phycocyanin included 6.2 ± 0.86 and 3.8 ± 1.06 μg/μL against ViDr, 6.90 ± 1.11 and 5.87 ± 3.43, respectively. Their cytotoxicity was low against normal cells. The expression of Bax gene was significantly increased in exposure of extract + phycocyanin and also C. colocynthis extract treatment. In addition, the combination of extract + phycocyanin and C. colocynthis extract significantly increased the expression of caspase 3 gene in CRC cell lines. ConclusionC. colocynthis extract, as well as the combination of extract + phycocyanin increased the expression of Bax and caspase 3 genes, indicating intervention in the apoptosis in cancer cells. Although the expression of Bax and Caspase 3 is a major anticancer mechanism of extract and phycocyanin, this finding and other mechanisms require further investigations.

L-Arginine protects mouse Leydig cells against T-2 toxin-induced apoptosis in vitro.

To explore the protective mechanism of L-arginine against T-2 toxin-induced apoptosis in mouse Leydig cells, we investigated whether L-arginine can prevent T-2 toxin-induced apoptosis in mouse Leydig cells and explored the underlying mechanisms. Leydig cells were isolated and cultured with control, T-2 toxin (10 nM), L-arginine (0.25, 0.5, and 1.0 mM), and T-2 toxin (10 nM T-2 toxin) + L-arginine (0.25, 0.5, or 1.0 mM) for 24 h. Cells and supernatants were harvested to examine proliferation of the cells, the apoptosis rate, activity of caspase-3 and mitochondria, and the gene expression levels of Bcl-2, Bax, PARP, and caspase-3. Results showed that proliferation and mitochondrial activity of Leydig cells were inhibited by administration of T-2 toxin. Bcl-2 gene expression levels was decreased, while the gene expression levels of Bax and PARP were increased, which could trigger mitochondria-mediated apoptosis, activate downstream caspase-3, and then increased caspase-3 at both activity and gene expression levels. The expression of the Bcl-2 gene was upregulated and the expression of Bax, caspase-3, and PARP gene were downregulated when L-arginine was added to the cultured cells. The results of this study showed that L-arginine could block T-2 toxin-induced apoptosis in mouse Leydig cells by regulating specific intracellular death-related pathways.

Apigenin inhibits proliferation of hepatocellular carcinoma cell by upregulation of cleaved caspases-3/8 and downregulation of pSTAT-3/pJAK-1/pJAK-2

Apigenin is a flavonoid nutrient and possesses antineoplastic actions. Cell proliferation inhibition, induction of apoptosis, cell cycle arrest and inhibition of cancer cell spread were assessed. HepG2 cells were divided into 3 groups: HepG2/control group, HepG2/DMSO group and HepG2/Apigenin group. HepG2 cells were cultured for 24, 48 and 72 h. Apigenin was supplemented on HepG2 (100 μM). Cell viability was assessed. qRT-PCR was applied for PARP, Bcl-2 and Bax genes expression. Western blot was estimated for cleaved caspases-3/8, HIF-α, and pSTAT-3/pJAK-1/pJAK-2 proteins. VEGF level was assessed by ELISA in cell culture supernatants. Finally, ultrastructure examination of cell culture pellet by transmission electron microscope (TEM) was determined. The results revealed that cleaved caspases-3/8 and Bax were significantly upregulated and the level of PARP, BcL-2, HIF-α, pSTAT-3, pJAK-1, pJAK-2 and VEGF were significantly downregulated in apigenin group compared to control groups.

The Effect of High Intensity Interval Training (HIIT) and Crocin Consumption on Doxorubicin-Induced Apoptotic Indices in the Muscle Tissue of Rats

Background: Apoptosis is one of the main means to remove undesirable cells that occurs in the eucaryotic and even procaryotic organisms. Objectives: This study aimed to examine the effect of 8 weeks of high-intensity interval training (HIIT) and crocin consumption on apoptotic factors in the soleus muscle of male rats exposed to doxorubicin. Methods: sixty male rats were selected and divided into five groups: 1) doxorbicin- control, 2) training, 3) crocin, 4) training + crocin, and 5) healthy control. Groups 1 to 4 received 2 mg/kg doxorubicin intraperitoneally; Groups 3 and 4 received daily 10 mg/kg crocin, and groups 2 and 5 received HIIT including high-intensity interval repetitions (80 to 90% of maximum speed) and low-intensity interval repetitions (30 to 40% of maximum speed) at two-minute alternations for eight weeks and for five days each week. Results: Doxorubicin had a significant effect on increasing Bax and caspase-3 and decreasing Bcl-2 (P ≤ 0.05). However, training had a significant effect on reducing Bax and increasing Bcl-2 (P ≤ 0.05). Crocin consumption had a significant effect on decreasing Bax and caspase-3 and increasing Bcl-2 (P ≤ 0.05). In addition, training and crocin consumption had interactive effects on decreasing Bax and increasing Bcl-2 (P ≤ 0.05). Conclusions: It appears that the combination of HIIT with crocin extract can be used to increase the expression of the Bcl-2 protein gene as an anti-apoptotic factor and decrease the expression of Bax gene as a stimulating factor in the soleus muscle of male rats exposed to doxorubicin.

Injury of procymidone on testis and sperm in the male adolescent mice

To investigate the damage of procymidone to penile, testis and sperm in adolescent male mice. 4-week-old male ICR mice were allocated randomly to the treatment groups and the control group, with 8 mice in each group. Procymidone was administered to mice by gavage with dose 50 mg/kg(low dose), 100 mg/kg and 200 mg/kg, while the control group was given only an equal volume of soybean oil. On the 11 th days, the mice were sacrificed by spinal dislocation, their body masses were weighed, and the anogenital distance(AGD), testicular volume and penis length were measured. Furthermore the weight of the testes, epididymis and penis were weighed, the organ coefficients were calculated, and then a testis on one side and 1/2 penis tissue were fixed in 4% paraformaldehyde and used for histology analysis. The testis on the other side and the remaining 1/2 penis were used to detect androgen content and Caspase-3, Caspase-9, Caspase-12 and Bax gene expression. At the same time, one epididymis was randomly selected to detect sperm motility, density and morphology. The weight, penis length and testicular volume of mice in each experimental group did not change significantly, while the weight of testes and epididymis and testicular and epididymal coefficients of mice in the high-dose group decreased significantly(P&lt;0. 05). The testis seminiferous tubules of every treatment group showed different degrees of degeneration, spermatogenic cell damage, and decreased number of luminal sperm. Meanwhile, medium and high dose treatment groups showed hyperplasia of testicular stromal cells. However, there was no significant pathologic change in penile tissue in every treatment group. The sperm density of mice in the middle and high dose groups was lower than that of the control group(P&lt;0. 05), and the sperm deformity rate in the two groups was significantly higher than that of the control group(P&lt;0. 01), while the sperm motility of every dose treatment group was lower than that of the control(P&lt;0. 05). The testosterone in the testis of the middle and high dose groups were high than the control(P&lt;0. 001), while there was no statistically significant difference of the testosterone among all the groups in the penis(P&gt;0. 05). In addition, the expression of pro-apoptotic genes Caspase-3, Caspase-9, Caspase-12 and Bax in testis tissues of the high-dose group were significantly increased(P&lt;0. 05). Procymidone can cause damage to the structure and function of testes, reduce sperm quality, and increase the expression of certain pro-apoptotic genes in adolescent male mice.

Effect of Artemisia absinthium ethanolic extract on oxidative stress markers and the TLR4, S100A4, Bax and Bcl-2 genes expression in the kidney of STZ-induced diabetic rats.

The present study was conducted to examine antidiabetic effects of Artemisia absinthium ethanolic extract [A.absinthium] and to investigate its effects on oxidative stress markers and the expression of TLR4, S100A4, Bax and Bcl-2 genes in the kidney of STZ-induced diabetic rats. Thirty six rats (weight 200-250g) were randomly divided into diabetes and control groups. Induction of diabetes was performed using STZ (55mg/kg.bw). Biochemical parameters and oxidative stress markers (SOD and MDA) were measured using spectrophotometry after 60days of treatment. The expression of TLR4, S100A4, Bax and Bcl-2 were analyzed by real-time PCR. One-way analysis of variance (ANOVA) and Bonferroni post hoc test were used to compare the data. Diabetes significantly impairs the serum fasting blood glucose (FBG), lipid profile, urea, creatinine and albumin. At the end of treatment with A.absinthium extract, these parameters were close to the normal range. The results showed that the A.absinthium extract significantly decreased the kidney expression of TLR4, S100A4, Bax and increased the expression of Bcl-2 and improved oxidative stress markers (SOD and MDA) in the kidney tissues of treated rats. Also, all of these beneficial effects of the A.absinthium were dose-dependent. The extract of A.absinthium possesses antidiabetic effects. A.absinthium decreased the expression of TLR4, S100A4, Bax and increased the expression of Bcl-2 and improved oxidative stress. Therefore, this herbal extract can be used as an adjuvant treatment for diabetic complications.

Enhancement of Cisplatin Cytotoxicity by Cu(II)-Mn(II) Schiff Base Tetradentate Complex in Human Oral Squamous Cell Carcinoma.

Oral squamous cell carcinoma (SCC) is one of the most predominant tumors worldwide and the present treatment policies are not enough to provide a specific solution. We aimed to assess the cytotoxic effect of Cu(II)–Mn(II) Schiff base tetradentate complex alone or in combination with cisplatin against squamous cell carcinoma cell line (SCCs) in vitro. Oral-derived gingival mesenchymal stem cells (GMSCs) were used as control. The cell viability was assessed by MTT assay. IC50 values were calculated. Evaluation of apoptosis and DNA damage were performed. In addition, the expression of pro-apoptotic and anti-apoptotic genes and proteins were tested. IC50 values indicated less toxicity of the Schiff base complex on GMSCs compared to cisplatin. Schiff base complex treatment resulted in up-regulation of p53 and Bax genes expression and down-regulation of Bcl2 gene expression in SCCs paralleled with increased protein expression of caspase-3 and Bax and down-regulation of Bcl-2 protein. Annexin V-FITC apoptosis kit showed a higher apoptotic effect induced by a Schiff base complex compared to the cisplatin-treated group. These effects were markedly increased on the combination of Schiff base and cisplatin. The present study established that Cu(II)–Mn(II) Schiff base tetradentate complex might induce a cytotoxic effect on SCCs cells via induction of the apoptotic pathway. Moreover, this Schiff base complex augments the anticancer effect of cisplatin.

Effect of coated carbon nanotubes with chitosan and cover of flaxseed in the induction of MDA-MB-231 apoptosis by analyzing the expression of Bax and Bcl-2

IntroductionFunctionalized carbon nanotubes have been used as molecular carriers in the cancer treatment. Medicine molecules bind to the surface functional groups of coated nanotubes or polymers. The aims of this research were to examine the effect of multilayer coated and uncoated carbon nanotubes containing flaxseed extract on MDA-MB-231 cell line and to study changes in induced apoptosis by measuring genes' expression of Bax and Bcl-2. Bcl-2 is anti-apoptotic gene and Bax is apoptotic gene. MethodsIn this study, MDA-MB-231 that is breast cancer cell line was cultured in DMEM medium as the culture media. The cells were divided into four equal groups, control group and three treatment groups. Cells were treated with different concentrations of flaxseed extract as medicine, chitosan-coated carbon nanotube and uncoated carbon nanotubes. The effects of the medicine were evaluated by MTT assay. Changes in the expression of Bax and Bcl-2 were analyzed by Real Time PCR. ResultsThe results showed that the expression level of Bax gene in chitosan-coated nanotubes carrying flaxseed and carbon nanotubes-containing flaxseed extract and crude carbon nanotubes and flaxseed extracts were respectively 9.125, 5.139, 1.576 and 3.151 that have significantly increased. Expression of Bcl-2 gene in chitosan-coated nanotubes containing flaxseed and carbon nanotube-coated nanotubes and crude carbon nanotubes and flaxseed extract were respectively −3.389, −2.915, −1.103 and − 2.604 that have significantly decreased. The results showed that flaxseed extract with carbon nanotube-chitosan nanoparticle carrier in a dose-dependent manner increased Bax gene expression and decreased Bcl-2 gene expression. ConclusionThe results showed that MDA-MB-231 cancer cells treated with carbon nanotube-chitosan nanoparticle and carrier of flaxseed extract had apoptosis and death due to increasing Bcl-2 and decreasing Bax. The use of flaxseed as a medicine and carbon nanotube-chitosan nanoparticle can be considered as a promising strategy in the treatment of breast cancer.

Characterization and apoptotic effect of copper nanoparticles biosynthesized from Ziziphus zizyphus leaf on human renal cell carcinoma A498 cells

Biological formulation of copper nanoparticles (Cu-Nps) in cancer treatment has augmented due to its beneficial and curative effects. Ziziphus zizyphus (ZZ) is a species of medicinal plant traditionally used for various ailments. Hence, this work deals with formulation of copper nanoparticles (Cu-NP’s) from Z. zizyphus leaf extracts (Cu-ZZ NP’s) and examine its effect on human renal carcinoma A498 cells. The characterization of the formulated Cu-ZZ NPs was done by spectroscopic and microscopic methods. The anti-cancer tests (MTT, ROS, MMP, Hoechst staining, comet, Real‐time PCR, and western blot) were used to evaluate the biological properties of Cu-ZZ NPs on the growth and proliferation of A498 cells. The cytotoxic activity of Cu-ZZ NPs resulted in potent toxicity of A498 cells. Further, Cu-ZZ NP’s triggered an intracellular ROS generation through mitochondrial pathways in the A498 cells, revealing a noteworthy cytotoxicity activity. Also, fluorescent microscopic studies using Hoechst staining, and comet analysis confirmed the occurrence of apoptosis in Cu-ZZ NPs treated A498 cells. Following, quantitative RT-PCR presented a negative regulation of Bcl-2 expression and positive regulation of Bid, Bax, and caspases (3 and 9) gene expression in Cu-ZZ NP’s treated A498 cells. To further justify, inhibition of mTOR/PI3K/AKT protein expression by Cu-ZZ NPs in A498 cells, lead to an inference that Cu-ZZ NPs induce apoptosis in A498 cells by inhibiting the mTOR, PI3K/Akt pathway.

Downregulation of Sirt1 is correlated to upregulation of p53 and increased apoptosis in epicardial adipose tissue of patients with coronary artery disease.

The higher expression level of p53 in epithelial adipose tissue (EAT) has previously been reported in atherosclerosis. Since we hypothesized that the expression of p53 is modulated by Sirt1, the aim of this study was to determine the expression levels of Sirt1 and p53 and to investigate their correlation to apoptosis in EAT of patients with coronary artery disease (CAD). Thirty-five patients with more than 50 % stenosis in at least one of the main coronary arteries were considered as CAD group while 29 patients with no clinical signs of atherosclerosis who underwent open-heart surgery for valve replacement were classified as control group. EAT biopsy samples were collected from all participants during surgery. Sirt1, p53, Bax, and Bcl-2 gene expression levels were determined in EAT by qRT-PCR and Western blotting was carried out to assess Sirt1 and p53 protein levels. Hematoxylin and eosin staining was used for histopathological analysis. mRNA and protein levels of Sirt1 in EAT were significantly lower in patients with CAD compared with control group, whereas CAD patients showed greater p53 gene and protein expressions. In addition, inverse correlations were observed between Sirt1 and p53 at both mRNA and protein levels. The Bax and ratio of Bax/Bcl-2 gene expressions were higher in CAD group, but no difference was observed in Bcl-2 expression. Histopathological analysis showed apoptotic bodies and infiltrated immune cells in EAT of CAD group. Our results suggest that the Sirt1-p53 axis may involve in atherosclerosis by promotion of apoptosis.

Aronia melanocarpa Ameliorates Adrenal Cytoarchitecture Against the Hexavalent Chromium-Induced Injury

Hexavalent chromium is a toxin that penetrates the cell, triggering reactive oxygen species (ROS) production. Aronia melanocarpa, due to its proanthocyanidins, anthocyanins, and phenolic acid contents, is a valuable antioxidant. The aim was to observe the influence of hexavalent chromium Cr(VI) on the adrenal gland, and if this impact can be recovered by the administration of A. melanocarpa. Accordingly, 36 rats were divided into six groups: control; Aronia; Cr receiving Cr(VI) in distilled water for 3 months; CrA receiving a mix of Cr(VI) and A. melanocarpa at 2.5% aqueous extract for 3 months; Cr2 receiving, for 3 months, Cr(VI) in distilled water, and next, for 1 month, only distilled water; and respectively, CrA2 receiving, for 3 months, Cr(VI) in distilled water, followed by 1 month of Aronia at 2.5% extract administration. The adrenal gland samples were examined toward histological and molecular assessment, and results were statistically analyzed (ANOVA). Hexavalent chromium induced changes in the adrenal cortex expressed by focal or diffuse hypertrophies, cytoplasmic vacuolization (due to lipidic accumulation), and cells’ shape and size alteration, including necrosis. These structural alterations were carried by Bax and Bcl2 gene expression: the Bax gene expression levels, increased significantly (p < 0.001) in all experimental groups, except the Aronia group, compared with control. In the Cr2, CrA, and CrA2 groups, notable reduction of Bax gene expression (p < 0.001) was reported compared with the Cr group. Regarding the Bcl2 gene expression (p < 0.001), a significant increase was observed in the experimental groups, compared with the control. The Bcl2 expression level had a similar pattern to Bax gene, consequently trying to compensate its overexpression. Aronia administered concomitantly, or after Cr(VI), diminished structural changes and expression of the studied genes, thus reducing the Bax/Bcl2 ratio and suggesting that the active ingredients from Aronia are capable of blocking apoptotic cascade induced by the pathway of Bax and Bcl2 proteins.

Protective Effects of Quercetin and Melatonin on Indomethacin Induced Gastric Ulcers in Rats.

Medications to prevent the development of NSAID-induced gastric ulcers have a large range of unpleasant side effects. Recent efforts have been focused on determining safer alternative nontoxic and natural forms of anti-ulcer treatments. Twenty-four male rats were divided into 4 groups: 1: control group that received no treatment; 2: the ndomethacin-treated group that received 20 mg/kg of indomethacin for 2 days to induce the development of gastric ulcers; 3: quercetin-treated group that in addition to the indomethacin treatment, received 50 mg/kg of quercetin 6 hours after and then daily for 14 days and; 4: the melatonin-treated group which received 20 mg/kg of melatonin 6 hours after each indomethacin treatment and then daily for 14 days. All drugs were administered orally. The following parameters were assessed in each group: mean ulcer index of gastric tissue, gastric acid volume and pH, oxidative stress markers: malondialdehyde (MDA), superoxide dismutase (SOD), glutathione peroxidase (GSH), inflammatory markers: PGE-2, TNF-α, and IL-10, nitric oxide (NO) levels and the relative gene expression of BAX, BCL-2 and COX-2 by real time PCR. Our findings revealed that the indomethacin-treated group had a significantly increased (p< 0.05) ulcer index, gastric acid volume, and elevated levels of stress, inflammatory, and apoptotic markers compared to controls. In the groups that received quercetin or melatonin, these factors were all significantly decreased (p< 0.05). Between quercetin and melatonin, there was no significant difference in their gastroprotective effect. Both quercetin and melatonin had protective antioxidant, anti-inflammatory and antiapoptotic activity against indomethacin-induced gastric ulcers.

Effect of the Cellular Extract of Co-cultured Lactobacillus Casei on BAX and Human β-Defensin 2 Genes Expression in HT29 Cells

Aims: Defensins are cysteine-rich antimicrobial cationic peptides and BAX is a proapoptotic gene that can cause cell death. This study aimed to investigate the effect of cellular extract of co-cultured Lactobacillus casei on the expression of BAX and human β-defensin 2 (hBD-2) genes in HT29 cells. Methods &amp; Materials: This experimental study was conducted in the Research Center for Pharmaceutical Nanotechnology of Tabriz University of Medical Sciences in 2017. The HT29 cell line was obtained from the Pasteur Institute of Iran, and cells were assessed using Microculture Tetrazolium Test (MTT) after culturing. DNA was extracted from the treated cells, and then the DNA ladder assay was carried out. After preparing cDNA, the expression levels of BAX and hBD-2 genes in the HT29 cell line were measured using a real-time Polymerase Chain Reaction (PCR) method. Findings: The results of the MTT assay indicated that Lactobacillus casei inhibited the proliferation of HT29 cells and induced apoptosis in these cells. Results of DAPI staining and DNA ladder assay obtained from treating HT29 cells by Lactobacillus casei showed qualitative changes in cell apoptosis. Moreover, realtime PCR results indicated that Lactobacillus casei bacteria significantly increased the expression of the hBD-2 gene in HT29 colon cancer cells within 12-24 hours (P= 0.023), while BAX gene expression showed no significant change in the first 24 hours (P= 0.37). Conclusion: The extract of Lactobacillus casei can be used to stimulate cancer cells to produce β-defensins, inhibit pathogens, prevent the stimulation of cellular signaling, and fight antibiotic-resistant bacteria.

Effect and mechanism of rapamycin on proliferation and apoptosis of human lung cancer cells

The purpose of this study was to investigate the effects of RAPA on the proliferation and the expression of p53, Bcl-2 and Bax proteins in cultured human small cell lung cancer (NCI-H446) cells, and to explore the possible mechanism of RAPA-treated NCI-H446 cells with different concentrations of RAPA-treated NCI-H446 cells. The proliferation of NCI-H446 cells in all groups was assayed by the CCK-8 method. FITC-Annexin V/PI double staining method was used to determine the apoptosis of NCI-H446 cells. The immunohistochemical SP method was used to detect the expression of p53, Bcl-2 and Bax. Expression of p53, Bcl-2 and Bax mRNA was detected by RT-PCR. The results showed that, after 48h treatment, the proliferation of NCI-H446 cells treated with 5ng/mL, 10ng/mL and 15ng/mL RAPA decreased significantly (P &lt; 0.05) and the proliferation inhibition rate increased significantly (P &lt; 0.05) compared with the control group, and the proliferation inhibition rate had a dose-dependent relationship with RAPA. Compared with the control group, the apoptosis rate of NCI-H446 cells treated with 5ng/mL, 10ng/mL and 15ng/mL RAPA increased significantly (P &lt; 0.05), and there was a dose-dependent relationship between the apoptosis rate and RAPA. The expression of Bcl-2 protein and mRNA was higher in the control group, while the expression of p53 and Bax protein and mRNA was lower. The expression of Bcl-2 protein and mRNA decreased and the expression of p53 and Bax protein and mRNA increased gradually with the increase of concentration and the prolongation of action time in 5ng/mL, 10ng/mL and 15ng/mL RAPA groups. In the control group, the intracellular Ca2+ concentration was constant, and there was no significant change with time; while in the 5ng / mL, 10ng / mL, and 15ng / mL RAPA group, the intracellular Ca2+ concentration in the RAPA group increased significantly after 12 h of administration (P &lt;0.05); After that, with the prolonged action time of the medicine, the intracellular Ca2+ concentration in the 5ng / mL, 10ng / mL, and 15ng / mL RAPA group decreased, but at 72h, the effect was 5ng / mL, 10ng / mL, and 15ng / mL RAPA. The intracellular Ca2+ fluorescence intensity in the group was still significantly higher than that in the control group (P &lt;0.05). In conclusion, RAPA can induce apoptosis of NCI-H446 cells by down-regulating Bcl-2 gene expression, up-regulating P53 and Bax gene expression, and increasing intracellular Ca2+ concentration and its apoptosis induction effect have timeliness and dose-effect.