Bcl-2 Gene Expression Levels Research Articles

Assessment of the Regulatory Effect of Novel Indole-Core-Based Compound on Apoptosis and Cell Survival of Acute Myeloid Leukemia Cell Line

Background: Heterocyclic compounds are generally introduced as important valuable sources of pharmacologically active compounds. Amongst these compounds, the indole is largely distributed within the bioactive molecules, containing antitumor agents. Due to their unique physiochemical and biological properties, indole and its derivatives have been used as privileged scaffolds for designing antitumor agents. Objective: The current experimental study aimed to evaluate the anti-cancer effects of a novel compound with indole-core-base on acute myeloid leukemia (AML) cells. Methods: Following being cultured, AML cells that had been multiplicities were treated by the demonstrated concentration of novel indole compounds (at doses of 100-300µg/mL) for 24 h. The percentage of living and dead cells was subsequently determined by trypan blue dye . The survival rate of treated cells was also examined by MTT assay. The calculated fold changes of the studied genes expression against β-actin were determined by real-time polymerase chain reaction (PCR) technique. The collected data were statistically analyzed by student t-test and repeated measure test. Results: Results showed that the intended novel indole-core-based derivative (C18H10N2F6O) followed both dose-dependent cytotoxic and anti-proliferative patterns on the AML cell line. The compound was able to induce apoptosis in 50% of the cells at the dose of 250µg/mL. Real-time PCR analysis indicated that in compound-treated cells the gene expression level of Bcl-2 has been downregulated, while Bax was upregulated, compared to untreated control cells. Conclusion: Despite the lack of knowledge in this regard, in this study, results of this leading mechanism(s) that may run by the compound indicated that indole3carbaldehyde derivative has cytotoxic effects on AML cells in a dose-dependent fashion.

Effect of moxibustion and acupuncture on gastric mucosal cell apoptosis and expression of NF-κB, Bcl-2 in chronic atrophic gastritis rats

To observe the effect of moxibustion and acupuncture on apoptosis in gastric mucosal cell and expression of NF-κB, Bcl-2 in chronic atrophic gastritis (CAG) rats, so as to explore its mechanisms underlying improvement of CAG. Forty-eight male SD rats were randomly divided into normal, model, moxibustion and acupuncture groups, with 12 rats in each group. The CAG model was established by gavage of N-methyl-N'-nitro-N-nitroso-guanidine (MNNG) solution and irregular diet for 12 weeks. Moxibustion or acupuncture was applied to "Zusanli" (ST36) and "Zhongwan" (CV12) for 15 min, once daily for two weeks in the moxibustion or acupuncture group. The histopathological changes of the gastric mucosa were observed by HE staining. The apoptosis index of gastric mucosa was measured by TUNEL method. The gene expression levels of NF-κB and Bcl-2 were detected by real-time PCR. After modeling, the color of gastric mucosa was pale and dark, with low folds and significant bleeding points. The glands in lamina propria were atrophied, arranged disorderly, and the numbers were significantly reduced, with inflammatory cells infiltrated. Those histopathological changes were evidently milder in the moxibustion and acupuncture groups. Compared with the normal group, the apoptosis index, NF-κB and Bcl-2 gene expression of the model group were significantly increased (P<0.01). After the treatment, the apoptosis index, NF-κB and Bcl-2 gene expression were significantly reduced in the moxibustion and acupuncture groups (P<0.05，P<0.01). The NF-κB expression was lower in the acupuncture group than that of moxibustion group (P<0.01). Moxibustion and acupuncture therapy can improve the injury of gastric mucosa in CAG rats, which may be associated with its function in down-regulating the expression of NF-κB and Bcl-2 genes in the gastric mucosa.

Design and synthesis of diosgenin derivatives as apoptosis inducers through mitochondria-related pathways.

Diosgenin (DSG) has attracted attention recently as a potential anticancer therapeutic agent due to its profound antitumor activity. To better utilize DSG as an antitumor compound, two series of DSG-amino acid ester derivatives (3a-3g and 7a-7g) were designed and synthesized, and their cytotoxic activities against six human cancer cell lines (K562, T24, MNK45, HepG2, A549, and MCF-7) were evaluated. The results obtained showed that a majority of derivatives exhibited cytotoxic activities against these six human tumor cells. Structure-activity relationship analysis revealed that the introduction of l-tryptophan to the C-3 position of DSG and the C-26 position of derivative 5 was the preferred option for these compounds to display significant cytotoxic activities. Among them, compound 7g exhibited significant cytotoxicity against the K562cell line (IC50=4.41μM) and was 6.8-fold more potent than diosgenin (IC50=30.04μM). Further cellular mechanism studies in K562cells elucidated that compound 7g triggered mitochondrial-related apoptosis by increasing the generation of intracellular reactive oxygen species (ROS) and decreasing mitochondrial membrane potential (MMP), which was associated with upregulation of the gene and protein expression levels of Bax, downregulation of the gene and protein expression levels of Bcl-2 and activation of the caspase cascade. The above results suggested that compound 7g might be considered a promising scaffold for further modification of more potent anticancer agents.

DP from Euphorbia fischeriana S. mediated apoptosis in leukemia cells via the PI3k/Akt signaling pathways

Ethnopharmacological relevanceEuphorbia fischeriana S. (E. fischeriana) is a classic Chinese herb with toxicity that is mainly used for cancer treatment and in insect repellent, anti-inflammatory and anti-edema applications (Liu et al., 2001). 12-Deoxyphorbol13-palmitate (DP), a tetracyclic diterpene monomer compound, was extracted from the roots of E. fischeriana by our research groups. AimPrevious studies found that DP could inhibit the proliferation of leukemia cells in vitro. However, the underlying mechanism of DP in leukemia is unknown. Hence, DP’s pharmacological effect on leukemia cells was investigated in this study. Materials and methodsDP was obtained from the Natural Medicine Chemistry Laboratory of Qiqihaer Medical University. In vitro, K562 cells and HL60 cells were incubated with DP or DP combined with LY294002 at different concentrations. Cell proliferation and apoptosis were detected by the relevant experimental methods. In vivo, nude mouse xenograft models were established by injecting K562 cells. DP was intraperitoneally administered to observe the influence on the growth of transplanted tumors. Gene detection and immunoblot analysis were performed to validate the mechanisms. ResultsThe cell counting kit-8 (CCK-8) assay proved that DP inhibited the growth of K562 and HL60 cells in a time- or dose-dependent manner. At 12 h, DP could induce apoptosis by Annexin V-FITC/propidium iodide (PI) dual labeling, loss of mitochondrial membrane potential (MMP), intracellular reactive oxygen species (ROS), acridine orange/ethidium bromide (AO/EB) staining and transmission electron microscopy (TEM) observation in K562 or HL60 cells. Furthermore, in an assay of gene and protein expression, we found that DP could downregulate the gene and protein expression levels of Bcl-2, upregulate the gene and protein expression levels of Bax and Bim, and downregulate the protein expression levels of PI3k, p-Akt, and p-FoxO3a. Moreover, the effects of DP on proliferation and apoptosis in K562 cells were enhanced by LY294002. Then, we tested the antitumor effects of DP in vivo. Nude mouse xenograft models were established by subcutaneously injecting K562 cells. We found that tumor volume was significantly decreased in DP-treated xenograft nude mice. Morphologic changes, apoptosis degree, and related gene and protein expression levels in transplanted tumor tissue of DP-treated nude mice were assessed by different experimental methods. ConclusionsThe in vivo and in vitro experimental results indicated that DP might inhibit the proliferation and induce the apoptosis of leukemia cells, which might be a result of suppressing the PI3k/Akt signaling pathways.

Cytotoxic effect of 6-Shogaol in Imatinib sensitive and resistant K562 cells.

Chronic Myeloid Leukemia (CML) is a clonal hematopoietic malignancy characterized by the formation of BCR-ABL fusion protein. Imatinib (IMA) is a BCR-ABL tyrosine kinase inhibitor (TKI), which exhibited a high rate of response for newly diagnosed CML patients. Emergence of IMA resistance considered as a major challenge in CML therapy. Recent studies reported the anti-cancer effect of natural extracts such as 6-Shogaol (6-SG) which is extracted from ginger and the mechanisms involved in targeting of cancer cells. In the present study, we aimed to explore the potential anticancer effect of 6-SG on K562S (Imatinib sensitive) and K562R (Imatinib resistant) cells. K562S and K562R cells were incubated with increasing concentrations of 6-SG (5μM- 50μM) to determine its cytotoxic and apoptotic effects. Cell viability and apoptosis were investigated with spectrophotometric MTT assay and flow cytometric Annexin V staining, respectively. The mRNA expression levels of apoptotic related genes (BAX and BCL-2) and drug transporter (MDR-1 and MRP-1) genes were evaluated with qRT-PCR. According to our results, 6-SG treatment inhibited cell viability, induced apoptosis in both K562S and K562R cells. Based on our RT-PCR results, 6-SG enhanced pro-apoptotic BAX gene and decreased anti-apoptotic BCL-2 gene expression levels significantly in both treated K562S and K562R cells. Furthermore, 6-SG increased MDR-1 mRNA expression level in K562S and K562R cells in comparison with their control counterparts. Whereas, 6-SG decrease MRP-1 mRNA expression level in K562S cells significantly. It is the first study that reveals the apoptotic effect of 6-SG in CML cell line and IMA resistance. Therefore, 6-SG treatment can be suggested as a promising strategy for CML therapy.

Interleukin-22 Attenuates Acute Pancreatitis-Associated Intestinal Mucosa Injury in Mice via STAT3 Activation

Background/AimsInterleukin-22 (IL-22) is an important cytokine maintaining homeostasis at barrier surfaces. In this study, the role of IL-22 in acute pancreatitis-associated intestinal injury was further explored.MethodsSevere acute pancreatitis (SAP) was induced by administration of L-arginine in Balb/c mice at different time gradients. Histopathological examinations were made in both the pancreas and small intestine. Furthermore, recombinant murine IL-22 (rIL-22) was administrated to L-arginine-induced SAP mice by intraperitoneal injection. The mRNA levels of IL-22R1, Reg-IIIβ, Reg-IIIγ, Bcl-2, and Bcl-xL were detected in the small intestine by real-time polymerase chain reaction, and protein levels of total and phosphorylated STAT3 were assessed via Western blot.ResultsCompared with normal control group, 72 hours of L-arginine exposure induced the most characteristic histopathological changes of SAP, evidenced by pathological changes and serum amylase levels. Meanwhile, significant pancreatitis-associated intestinal mucosa injury was also observed. The gene expression levels of antimicrobial proteins Reg-IIIβ, Reg-IIIγ and anti-apoptosis proteins Bcl-2, Bcl-xL were downregulated in small intestine. Furthermore, L-arginine-induced SAP was attenuated by rIL-22 treatment. Importantly, pancreatitis-associated intestinal mucosa injury was also ameliorated, reflected by improved pathological changes and significant increase in gene expression levels of Reg-IIIβ, Reg-IIIγ, Bcl-2 and Bcl-xL. Consistently, serum amylase levels and mortality were decreased in mice treated with rIL-22. Mechanistically, the upregulated expressions of these protective genes were achieved by activating STAT3.ConclusionsExogenous rIL-22 attenuates L-arginine-induced acute pancreatitis and intestinal mucosa injury in mice, via activating STAT3 signaling pathway and enhancing the expression of antimicrobial peptides and antiapoptotic genes. (Gut Liver 2021;15-781)

Evaluation of cytotoxicity and anticancer activity of kombucha and doxorubicin combination therapy on colorectal cancer cell line HCT-116.

BACKGROUND:Colorectal cancer is the third leading to death type of cancer in the world. The therapeutic guideline varied between different methods. As the main therapeutic guideline is chemotherapy, recent studies had shown utilization of natural products in combination with conventional medication, elevate the efficiency of chemotherapeutic methods. Kombucha is a traditional beverage obtained from the fermentation of green tea as a rich source of flavonoid medicinal plant. This study aimed to evaluate the natural potential of combination therapy of this natural product with doxorubicin as a chemotherapeutic agent.MATERIALS AND METHODS:The study was performed as in vitro evaluation of biological activity of kombucha on HCT-116 cell line (human colon cancer cell line). The cytotoxic effect of different kombucha beverages (fermented green tea) in comparison with green tea extract was evaluated by dimethylthiazolyl tetrazolium bromide (MTT) assay. In the next step, anticancer activity of doxorubicin as a general guideline chemotherapeutic agent in combination with kombucha was evaluated by cell cycle analysis and apoptosis assay flow cytometry. Apoptotic genes expression pattern was determined using real-time polymerase chain reaction. The experiments were designed in three independent replications and statistically analyzed using SPSS software.RESULTS:The results show that kombucha compared with the green tea extract caused more (1.2 fold) early apoptosis induction and G0/G1 phase arrest. Moreover, kombucha increased the expression levels of p21, p53, and B-cell leukemia/lymphoma 2 (Bcl-2)-associated X protein genes (2, 2.5, and 1.5 fold, respectively) while it decreased Bcl-2 gene expression level (5–8 fold) compared with doxorubicin alone. Combination of kombucha with doxorubicin shows 2-fold increased G0/G1 phase compared with the doxorubicin treatment.CONCLUSION:This result indicated that kombucha caused boosted anticancer activity of doxorubicin agent. These findings suggest that kombucha may be has an assistor and useful role in colorectal cancer treatment align with chemotherapy.

Impactos de la Quercetin y Mercaptothion en los parámetros reproductivos de rata macho

Quercetin is an herbal polyphenol with valuable antioxidant properties. Mercaptothion is categorized as organophosphates which can generate free radicals and induce male fertility disorders. This study was aimed to assess the impacts of Quercetin against destruction of male fertility parameters induced by Mercaptothion. 64 male rats were randomly assigned into 8 groups; control, and Mercaptothion (250 mg/kg) groups; Quercetin groups (7.5, 15, and 30 mg/kg) and Mercaptothion + Quercetin (7.5, 15, and 30 mg/kg). Treatments were administered intraperitoneally (Mercaptothion) and orally (Quercetin) daily for 65 days. The sperm parameters, testis malondialdehyde (MDA), total antioxidant capacity (TAC), testosterone level and height of germinal layer were evaluated. Expressions of p53, caspase-3, Bax, and Bcl-2 were measured through real-time quantitative PCR. Values of all factors were reduced significantly except the MDA level (which increased) in Mercaptothion group compared to the control group (p&lt;0.001). Studied criteria in groups of Quercetin and Quercetin + Mercaptothion in whole doses increased significantly except MDA level (which reduced) compared to the Mercaptothion group (p&lt;0.001). Also, downregulated levels of p53, caspase-3, and Bax genes and unregulated levels of Bcl-2 gene expression were detected in control and the sixth treatment groups significantly in Quercetin group compared to the Mercaptothion group (p&lt;0.001). No significant alterations were detected in Quercetin groups compared to the control group (p&gt;0.05). Quercetin reduced toxic effects of Mercaptothion on male fertility parameters.

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.

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.

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.

Lycopene mitigates experimental colitis in rats by inhibiting oxidative stress-mediated inflammation and apoptosis

Objective: The potential protective and beneficial effects of Lycopene (LYC) in a rat model of acetic acid-induced ulcerative colitis (UC) were evaluated. Methods: Forty male albino rats were divided into five equal groups. Group I: (Control normal) rats received no drugs. Group II: (Early ulcerative colitis): rats received 2 ml (3% v/v) glacial acetic acid intracolonially at 21 th day of experiment and sacrificed 3 days later. Group III:(Early ulcerative colitis + Lycopene protected) rats received Lycopene (10 mg/kg body weight/day, orally) for 21 days prior to acetic acid administration. Group IV: (Late ulcerative colitis) rats received acetic acid similar to group II for 3 successive days and sacrificed after 21 days. Group V: (Late ulcerative colitis+ Lycopene treated) rats first administered with acetic acid then after 3 days Lycopene was administered for 21 days. Results: A significant increase in L-Malondialdehyde (L-MDA) with marked decrease in reduced glutathione (GSH) level and Catalase (CAT) activity were observed in colon tissue of UC-induced rats. Additionally, a significant up-regulation of nuclear factor kappa B (NF-κB), caspase-3 and Transforming growth factor-β1 (TGF-β1) and down-regulation of Interleukin -10 (IL-10) and Beta cell lymphoma-2 (Bcl-2) gene expression levels of were observed in colon of UC induced rats. However, a significant depletion of colon tissue L-MDA and down-regulation of NF-κB, caspase-3 and TGF-β1 in addition to marked increase in GSH concentration and CAT activity and up-regulation of IL-10 and Bcl-2 were observed after LYC treatment. Conclusions: Lycopene has powerful antioxidants, anti-inflammatory and anti-apoptotic effects against ulcerative colitis.

Combined treatment with silver graphene quantum dot, radiation, and 17‐AAG induces anticancer effects in breast cancer cells

We aimed to investigate the possible anticancer effects of radiation in combination with 17-allylamino-17-demethoxy geldanamycin (17-AAG) and silver graphene quantum dot (SQD) in breast cancer (BC) cells. MCF-7 BCcells treated with, or without, different concentrations of 17-AAG and synthesized SQD and cellular viability detected. The growth inhibitory effects of low concentrations of 17-AAG with minimally toxic concentration of SQD in combination with 2 Gy of X-ray radiation were examined. The apoptosis induction assessed by acridine orange/ethedium bromide staining. Likewise, the levels of lactate, hydrogen peroxide (H2 O2 ), nitric oxide (NO) were evaluated. The relative gene expression levels of Bax and Bcl-2 were detected by real-time polymerase chain reaction and the Bax/Bcl-2 expression ratio was determined. Moreover, the protein expression of epidermal growth factor receptor (EGFR) was assessed by western blot analysis. Treatment with low concentrations of 17-AAG and SQD at a minimally toxic concentration promoted inhibition of BC cell growth and induced apoptosis. In addition, significant reduction in cell viability was seen in triple combination versus all double and single treatments. Indeed 17-AAG and SQD in combined with radiation significantly increased the H2 O2 and NO versus single and double treated cases. In addition, triple combination treatment showed decreased lactate level in compared tomonotherapies. EGFR protein expression levels were found to decreased in all double and triple combined cases versus single treatments. Additionally, in double and triple treatments, Bax/Bcl2 ratio were higher in compared to single treatments. Treatment with low concentrations of 17-AAG and SQD at a minimally toxic concentration tends to induce anticancer effects and increase the radiation effects when applied with 2 Gy of radiation versus radiation monotherapy.

Trichinella spiralis muscle larvae excretory/secretory products trigger apoptosis and S-phase arrest of the non-small-cell lung cancer line A549

Trichinella spiralis (T. spiralis) muscle larvae (ML) excretory/secretory products (ESPs) are antitumor substances extracted from the culture medium of T. spiralis ML. The ESPs inhibit tumor growth and induce tumor cell apoptosis. To explore the effects of these products on the non-small-cell lung cancer (NSCLC) line A549, logarithmically growing A549 cells were co-cultured with different concentrations of T. spiralis ML ESPs for 24, 36 and 48 h. Our results showed that T. spiralis ML ESPs significantly inhibited A549 cells proliferation, which was dose-and time-dependent. To evaluate the inhibition by T. spiralis ML ESPs of the growth of A549 cells, we assayed their apoptosis and cell-cycle distribution by flow cytometry (FCM). To determine whether ESPs induced apoptosis of A549 cells via the mitochondrial pathway, we evaluated the levels of mitochondrion-related factors by Western blotting. The FCM indicated a clear trend toward apoptosis of A549 cells co-cultured with ESPs for 24 h. The cells were blocked in S-phase. Western blotting revealed that the expression levels of the genes encoding Bax, caspase-3, and caspase-9 increased (compared to a control group), and the Bcl-2 gene expression level decreased. Our results suggest that T. spiralis ML ESPs induce apoptosis of the NSCLC line A549 via the mitochondrial pathway; the cells become arrested in S-phase. This may explain the antineoplastic activity of T. spiralis ML ESPs.

Secreted Chemicals From Probiotic Bacteria Potentiate Th1 Pattern of Immune Cells and Apoptosis Induction in Breast Cancer and Gastric Adenocarcinoma Cell Lines

Cancer is one of the main causes of mortality. Therapeutic importance in some diseases, especially cancers has led to considering the anti-cancer effects of probiotics more than ever. Probiotics have stimulatory effects on the immunity and suppressor effects on cancer activity. Various studies report that probiotic bacteria have cytotoxicity effects on different types of solid tumors. In this research, we evaluated the synergic effect of secretory chemicals of Lactobacillus and Bifidobacterium bacteria on cytotoxic potency of breast and gastrointestinal cancer cell lines. Bifidobacterium bifidum and Lactobacillus acidophilus were cultured in MRS broth. Cytotoxicity assay was performed on AGS, MCF-7 cell lines, and peripheral mononuclear cells after treatment with bacteria extractions by MTT test. β-actin, Bcl2, Bax, TNF-α, and IFN-γ gene expression levels were evaluated in treated cell lines by real-time PCR method. The result of cytotoxicity assay showed that the obtained extraction from the bacterial cultures has a higher killing effect on cancer cells compared with normal cells. Results of gene expression indicated a significant increase in Bax and Bcl2 gene expression in cancer cells. TNF-α gene expression increased significantly as compared with the control group. The obtained extraction from probiotics culture can induce death in cancer cells through the apoptosis mechanism and improve the cellular immune response, i.e., T helper 1 cytokine profile.

Effect of Fucoxanthin Administration on Thyroid Gland Injury Induced by Cadmium in Mice.

The intervention effect of fucoxanthin, which is reportedly a powerful antioxidant, on cadmium-induced thyroid damage in mice was evaluated. Animals (N = 120) were divided into control group (given pure water, N = 20) and CdCl2-exposed group (given CdCl2 orally at a dose of 30 mg/kg body weight (bw)/day for 30 days, N = 100). Besides, the CdCl2-exposed group was divided into the following 5 groups (N = 20) to evaluate the intervention effect of fucoxanthin: (1) negative control group (NCG; animals were supplied with pure water); (2) positive control group (PCG; animals were supplied with 50 mg/kg bw/day thyroid tablets. Thyroid tablets are made from the thyroid glands of pigs, cattle, sheep and other food animals (the main components of T4). But because they are extracts of the thyroid glands, they may contain a small amount of T3. The specific amount of T3 is unknown.); (3) low fucoxanthin concentration group (F1; animals were supplied with 10 mg/kg bw/day fucoxanthin); (4) medium fucoxanthin concentration group (F2; animals were supplied with 25 mg/kg bw/day fucoxanthin); (5) high fucoxanthin concentration groups (F3; animals were supplied with 50 mg/kg bw/day fucoxanthin). A 14-day treatment was conducted for these animals. The levels of T4, T3, MDA, ascorbate peroxidase (APX) and catalase were measured, and the expression levels of Bax, Bcl-2, ERK1, ERK2, MEK1, eIf2α, p-eIf2α, GRP78 and GRP94 genes were determined using real-time reverse transcriptase-polymerase chain reactions (RT-PCR). In addition, tissue histopathology and ultrastructure were recorded and analysed. We found that the injection of cadmium chloride (CdCl2) decreased serum T4 and T3 levels to 27.10 ng/ml and 837.74 pg/ml, respectively. In addition, CdCl2 intoxication induced oxidative stress, structural abnormalities and apoptosis in thyroid follicles. Our results showed that the treatment of CaCl-exposed mice with 25-50 mg/kg bw/day fucoxanthin appreciably decreased oxidative stress and apoptosis induced by CdCl2 and restored the microstructural and ultrastructural organisations of the thyroid gland towards normalcy. Compared with the negative control group, fucoxanthin treatment groups showed significant increase in T4 and T3 levels (52.17 ng/ml and 1669.18 ng/ml, respectively; P < 0.05), relieved oxidative stress by decreasing malondialdehyde level and increasing catalase and APX levels and increased apoptosis inhibition through inhibiting the ERK1/2 pathway and preventing endoplasmic reticulum stress in thyroid follicular epithelial cells. Herein, our study provides evidence of the protective effects of fucoxanthin supplementation against thyroid damage and suggests therapeutic potential of this pigment in cases of Cd intoxication and hypothyroidism.

The immunoreaction and antioxidant capacity of juvenile blunt snout bream (Megalobrama amblycephala) involves the PI3K/Akt/Nrf2 and NF-κB signal pathways in response to dietary methionine levels

A 75-day rearing trail was designed to evaluate the immunoreaction and antioxidant capacity of juvenile blunt snout bream in response to dietary methionine levels. Three practical diets were extruded to feed juveniles with graded methionine levels (0.40%, 0.84% and 1.28% dry matter). The data indicated that the plasma concentrations of immunoglobulin M (IgM), complement component 3 (C3) and glutathione (GSH) in the 0.84% methionine diet were markedly upper than those in the 0.40% group (P < 0.05). The activities of plasma antioxidant parameters involving catalase (CAT), total superoxide dismutase (T-SOD), total antioxidant capacity (T-AOC) and glutathione peroxidase (GPx) were significantly increased by the 0.84% diet compared with the 0.40% diet, whereas plasma alanine aminotransferase (ALT) and malondialdehyde (MDA) levels were significantly induced by 0.40% methionine (P < 0.05). Compared with the 0.40% group, 0.84% dietary methionine dramatically upregulated the mRNA expression levels of protein kinase B (Akt), phosphoinositide 3-kinase (PI3K) and nuclear factor erythroid 2-related factor 2 (Nrf2) pathway related genes including CAT, manganese superoxide dismutase (Mn-SOD), heme oxygenase 1 (HO-1) and glutathione peroxidase-1 (GPx-1) in the kidney and liver, and downregulated Kelch-like ECH-associated protein 1 (Keap1) mRNA levels (P < 0.05). Compared with the 0.40% group, the 0.84% dietary methionine strikingly suppressed the mRNA levels of renal and hepatic nuclear factor-kappa B (NF-κB) and pro-inflammatory cytokines (interleukin 1β (IL-1β), tumor necrosis factor-α (TNF-α), interleukin 6 (IL-6)), however, improved the mRNA expression levels of anti-inflammatory cytokines involved renal and hepatic transforming growth factor-β (TGF-β) and hepatic interleukin 10 (IL-10) (P < 0.05). Renal IL-10 and interleukin 8 (IL-8) mRNA expression levels were not markedly influenced by experimental diets (P > 0.05). Dietary methionine (0.84%) significantly upregulated renal and hepatic heat stress protein 70 (Hsp70), renal B-cell lymphoma-2 (Bcl-2) gene expression levels compared with the 0.40% diet (P < 0.05). In a word, the data represented that 0.84% dietary methionine could enhance the immune and antioxidant capacity of this fish species by inducing PI3K/Akt/Nrf2 pathway and inhibiting NF-κB pathway.

Study of Anticancer Activity of Pratensein and Pratensein Glycoside Isolated from Cuscuta kotchiana

In the present paper, we demonstrate that extract of Cuscuta kotchiana is able to inhibit in vitro proliferation of two human breast cancer cell lines, MCF-7 and MDA-MB-231. The expression levels of p53, bcl-2, caspase-3 and bax genes at the mRNA and protein levels were evaluated using quantitative Real Time PCR and western blot analysis. The most active fractions of C. kotchiana were detected by NMR as pratensein and pratensein glycoside. The cytotoxic activity of pratensein glycoside was significantly more than pratensein. The expression level of bcl2 gene was decreased in cancer cells treated by both compounds at CC50 concentrations. But the expression levels of caspase-3, p53 and bax genes were increased in treated cancer cells. In conclusion, all the data demonstrated that the glycoside form of pratensein is important agent in inducing apoptosis in human breast cancer cells.

Antioxidant effects of hydro-alcoholic extract of Ashrasi date palm in hepatotoxicity caused by mercuric chloride administration

Introduction: date palm (ADP) is a flowering plant type of the palm family which has strong antioxidant properties. Mercuric chloride (MC) is a complex substance that generates oxidative stress. This study was designed to investigate the protective effects of ADP on the , inflammatory, oxidative and apoptotic changes induced by MC.&#x0D; Methods: Fifty-six male rats were randomly allocated into 8 groups (n = 7) as follows: Group 1- control, 2- MC (50 mg/kg), 3-5: ADP (30, 90, and 270 mg/kg), and 6-8: MC+ADP. The rats were orally and administrated with the respective agents for 5 weeks. Extracts of ADP were prepared and screening was conducted. Nitric oxide (NO), lipid (LP), and Ferric Reducing Ability of Plasma (FRAP) assays were conducted to assess the antioxidant index. The levels of p53, Bcl2 and Bax gene expression were assessed by real-time PCR. Cytokines involved in inflammation were evaluated by enzyme-linked assay (ELISA). Liver enzymes were also measured using biochemical methods. changes (e.g. in hepatocyte (HD) and central hepatic vein (CHV)), and apoptosis cell index were evaluated by light and fluorescent microscopy methods.&#x0D; Results: MC treatment significantly increased all parameters described in the Methods section (except for FRAP level and Bcl2 expression level, which were both decreased) in the MC group, when compared to the control group (P &amp;lt; 0.05). Also, all indices were significantly decreased in the ADP and ADP+MC groups (except for FRAP level and Bcl2 expression level, which were increased), when compared to the MC group (P &amp;lt; 0.05).&#x0D; Conclusion: These findings revealed that ADP extract can serve as a potent antioxidant which effectively attenuates the adverse effects of MC on the liver through the activation of antioxidant mechanisms and regeneration of modifications.

Minocycline attenuates testicular damages in a rat model of ischaemia/reperfusion (I/R) injury.

Testicular torsion is a serious urological disease leading to testicular damage. This study aimed to assess the effect of minocycline on testicular ischaemia/reperfusion (I/R) injury caused by testicular torsion/detorsion. Male adult Wistar rats (n=32) were assigned into four groups of sham, I/R, I/R+minocycline and minocycline. I/R injury was induced by two sets of surgical operations, including the rotation of the left testis (720°, counterclockwise), followed by detorsion after 4hr. The administration of minocycline was carried out 30min before detorsion and then continued for 8weeks. At the end of the 8th week, rats were killed and sampling was done. Johnson's score, the height of seminiferous tubule epithelium, the mean seminiferous tubule diameter, as well as biochemical parameters, SOD, GPx and CAT, were significantly enhanced in the I/R+minocycline group compared with the I/R group. The administration of minocycline led to a marked decrease in expression levels of Caspase-3, Bax, IL-1β and TNF-α genes, and a remarkable increase in expression levels of Bcl-2, 3β-HSD and 17β-HSD3 genes compared with the I/R group. Administration of minocycline could also reduce the rate of germ cell apoptosis (TUNEL staining). Hence, minocycline was useful in the management of testicular torsion/detorsion.