Shibin Feng

Deoxynivalenol induces apoptosis in PC12 cells via the mitochondrial pathway.

Deoxynivalenol (DON) has broad toxicity in animals and humans. In this study the impact of DON treatment on apoptotic pathways in PC12 cells was determined. The effects of DON were evaluated on (i) typical indicators of apoptosis, including cellular morphology, cell activity, lactate dehydrogenase (LDH) release, and apoptosis ratio in PC12 cells, and on (ii) the expression of key genes and proteins related to apoptosis, including Bcl-2, Bax, Bid, cytochrome C (Cyt C), apoptosis inducing factor (AIF), cleaved-Caspase9, and cleaved-Caspase3. DON treatment inhibited proliferation of PC12 cells, induced significant morphological changes and apoptosis, promoted the release of Cyt C and AIF from the mitochondria, and increased the activities of cleaved-Caspase9 and cleaved-Caspase3. Bcl-2 expression decreased with increasing DON concentrations, in contrast to Bax and Bid, which were increased with increasing DON concentration. These data demonstrate that DON induces apoptosis in PC12 cells through the mitochondrial apoptosis pathway.

Immune regulation mechanism of Astragaloside IV on RAW264.7 cells through activating the NF-κB/MAPK signaling pathway

&NA; The present study was designed to investigate the effects of Astragaloside IV (ASIV) on the immune functions of RAW264.7 cells. Compared with control group, the concentrations of interleukin 1&bgr; (IL‐1&bgr;), tumor necrosis factor &agr; (TNF‐&agr;) and nitric oxide (NO) were higher in the 100 &mgr;g/mL ASIV‐treatment group. The interleukin 6 (IL‐6) concentration was significantly higher in the 50 and 100 &mgr;g/mL ASIV‐treatment groups. The relative mRNA expression levels of IL‐1&bgr;, TNF‐&agr; and inducible nitric oxide synthase (iNOS) were significantly higher in the 50 and 100 &mgr;g/mL ASIV‐treatment groups. The relative mRNA expression levels of IL‐6 in the 100 &mgr;g/mL ASIV‐treatment group were significantly higher. In contrast, the relative mRNA expression levels of interleukin 4 (IL‐4) and IL‐6 markedly reduced in ASIV‐treatment groups. Furthermore, ASIV promoted the secretion of CD40 and CD86 and increased the number of cells in G2/M phase. The apoptosis of RAW264.7 cells was decreased in ASIV‐treatment groups. The protein levels of cyclin D1, CDK4 and CDK6, p50 and p‐p65 increased in a dose‐dependent manner. The ratio of p50/&bgr;‐actin was significantly higher in the 50 and 100 &mgr;g/mL ASIV‐treatment groups, and p‐p65/p65 was significantly higher in the 25, 50 and 100 &mgr;g/mL ASIV‐treatment groups. The phosphorylation levels of p38, ERK and JNK increased, and the protein expression of total p38, ERK and JNK decreased in a dose‐dependent manner. These effects of ASIV were alleviated by PDTC. ASIV enhances the immune function of RAW264.7 cells by activating the NF‐&kgr;B/MAPK signaling pathway. HighlightsASIV activated the NF‐&kgr;B/MAPK signaling pathway in RAW264.7 cells.ASIV promoted the secretion of CDs and increased the number of cells in G2/M phase.ASIV activated the NF‐&kgr;B/MAPK to enhance immune functions in RAW264.7 cells.

Andrographolide Inhibits Inflammatory Cytokines Secretion in LPS-Stimulated RAW264.7 Cells through Suppression of NF-κB/MAPK Signaling Pathway

Andrographolide, the main active component extracted from Andrographis paniculata (Burm.f.) Wall. ex Nees, exerts anti-inflammatory effects; however, the principal molecular mechanisms remain unclear. The objective of this study was to investigate the molecular mechanisms of Andrographolide in modifying lipopolysaccharide- (LPS-) induced signaling pathway in RAW264.7 cells. An in vitro model of inflammation was induced by LPS in mouse RAW264.7 cells in the presence of Andrographolide. The concentration and expression levels of proinflammatory cytokines were determined by an enzyme-linked immunosorbent assay (ELISA) and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. The nuclear level of NF-κB was measured by an electrophoretic mobility shift assay (EMSA). The expression levels of NF-κB, p38, ERK, and JNK were determined by western blot. Andrographolide dose-dependently inhibited the release and mRNA expression of TNF-α, IL-6, and IL-1β in LPS-stimulated RAW264.7 cells. The nuclear level of p65 protein was decreased in Andrographolide treatment group. Western blot analysis showed that Andrographolide suppressed LPS-induced NF-κB activation and the phosphorylation of IkBa, ERK1/2, JNK, and p38. These results suggest that Andrographolide exerts an anti-inflammatory effect by inhibiting the activation of NF-κB/MAPK signaling pathway and the induction of proinflammatory cytokines.

Effects of β-conglycinin on growth performance, immunoglobulins and intestinal mucosal morphology in piglets

One of the main causes of allergic reactions in young animals is β-conglycinin, an antigenic glycoprotein found in soya beans. Therefore, the objective of the study was to investigate the effects of a prior immunisation with β-conglycinin on growth performance, serum immunoglobulin levels and intestinal histology in piglets. Forty piglets (7 d of age) were randomly divided into four groups of ten piglets each. Piglets of Groups Im and Im+S were immunised twice by hypodermic injection with β-conglycinin at 500 μg/kg body weight (BW) at day 7 and 21 of age. At day 23, Groups Im+S and S were intramuscularly injected with 5000 μg β-conglycinin per kg BW. The piglets of Group C received a physiological saline solution by hypodermic injection. All piglets were weaned at the age of 23 d and blood samples were taken on days 7, 21 and 35. At the end of the trial, five piglets per group were slaughtered and the intestine was collected for evaluating mucosal histology. Compared to Group C, in Group S the average daily gain (ADG), feed intake and gain:feed ratio were decreased (p < 0.01), and serum levels of IgG and IgE were increased (p < 0.01). Furthermore, in this group the structure of duodenal and jejunal mucosa was severely damaged. But in Groups Im and Im+S the ADG was increased (p < 0.05), serum IgE levels were decreased (p < 0.01) and the intestinal mucosa was not damaged. The results suggest that prior immunisation with β-conglycinin can increase ADG and serum IgG levels and decrease serum IgE levels. Therefore, this method is also potentially able to protect the structural integrity of the intestinal mucosal epithelia and alleviate allergic reactions in piglets.

Treatment of inflammatory bowel disease via green tea polyphenols: possible application and protective approaches

Inflammatory bowel disease (IBD) is a collection of inflammatory conditions of colon and small intestine which affect millions of individuals worldwide and the prevalence amount is on the rise. The organ failure as well as loss of tissue function is because of the inflammatory reaction which is the major contributor of tissue healing leading to lifelong debilitation. To stop the tough consequences of inflammation every patient pursues alternative therapy to relieve symptoms. Green tea polyphenols (GTPs) play significant roles in down regulating signaling pathways because GTPs exert effective antioxidant properties and regulate Toll-like receptor 4 (TLR4) expression via certain receptor, inhibited endotoxin-mediated tumor necrosis factor alpha (TNF-α) production by blocking transcription nuclear factor-kappa B (NF-kB) activation and upstream of mediated I kappa B kinase complex pathway activities, as well as intrusion with the flow of cytokines and synthesis of cyclooxygenase-2 (COX-2). This article highlights the green approach regarding the defensive effects of GTP review-related studies concerning the contrary effects and the key therapeutic targets application of GTPs in biomedical field to treat inflammatory bowel disease (IBD) and its complications.Graphical abstract.

A sensitive immunochromatographic assay using colloidal gold–antibody probe for rapid detection of fumonisin B1 in corn

ABSTRACT A sensitive immunochromatographic assay (ICA) using a colloidal gold–antibody probe for the rapid detection of fumonisin B1 (FB1) in corn samples was developed. The colour density of the test line correlated with the concentration of FB1 in the range 2–40 ng ml–1 by the assay, and the detection limit for FB1 was 2 ng ml–1. The linear range for FB1 was 50–1000 µg kg–1, and the visual limit detection of the test was 1000 µg kg–1 in corn samples. The ICA to detect FB1 is sensitive, specific and rapid. Specific anti-FB1 monoclonal antibody (mAb) and FB1-ovalbumin (FB1-OVA) conjugate antigen were prepared. FB1 mAb, labelled with colloidal gold, was used as the probe on the immunochromatographic strip. FB1-OVA and goat-anti-mouse IgG were coated onto a nitrocellulose (NC) membrane as test lines and control lines, respectively. FB1 in samples will competitively combines the FB1 mAb with the FB1-OVA in an NC membrane and the results are directly observed by the colour of the detection and quality control lines. The concentrations of FB1 mAb labelled with colloidal gold, detecting antigen and goat anti-mouse IgG, were optimised. The results indicate that the test strip is specific for FB1, with no cross-reactivity to other toxins. The strip assay for FB1 was simple, only needing one step without complicated assay performance and expensive equipment, and the total time for visual evaluation was less than 10 min. A survey of 24 corn samples from Hefei, China, was performed with the test strip and HPLC, and the detection results showed that the developed ICA and the HPLC were in excellent agreement. Hence, the developed ICA can be used as a method for rapid detection of FB1 in corn samples.

Chicken uric acid elimination via the uric acid transporters BCRP and MRP4 in the liver, kidneys, and intestines

Breast cancer resistance protein (BCRP) and multidrug resistance protein 4 (MRP4) are involved in uric acid excretion in humans and mice. Despite evidence suggesting that chicken renal proximal tubular epithelial cells participate in uric acid secretion, the roles of BCRP and MRP4 in chickens remain unclear. This study evaluated the relationship between chicken BCRP and MRP4 expression and renal function in the liver, kidneys, and intestines. Sixty 20-day-old Isa brown laying hens were randomly divided into four groups: a control group (NC) and groups provided with sulfonamide-treated drinking water (SD), a diet supplemented with fishmeal (FM), and an intraperitoneal injection of uric acid (IU). Serum uric acid, creatinine, and blood urea nitrogen (BUN) levels were significantly higher in the SD and IU groups than in the NC group. BCRP and MRP4 levels in the SD and IU groups were significantly increased in the kidneys and ileum and decreased in the liver. In the FM group, BCRP and MRP4 were significantly increased in the kidneys and slightly increased in the ileum. These results demonstrate that chicken BCRP and MRP4 are involved in renal and intestinal uric acid excretion. When renal function is impaired, serum uric acid increased and BCRP and MRP4 in the liver, kidneys, and ileum exhibit compensatory increases; when renal function is normal, serum uric acid changes have no effect on ileum BCRP and MRP4 expression. Therefore, this study may provide the references to the uric acid regulation in human.

Deoxynivalenol induces toxicity and apoptosis in piglet hippocampal nerve cells via the MAPK signaling pathway

Deoxynivalenol (DON) is a mycotoxin capable of producing a variety of toxic effects in human and animals. In this study, the effect of DON treatment on cytotoxicity and apoptotic pathways in piglet hippocampal nerve cells (PHNCs) was determined. The effects of DON on cellular morphology, cell activity, lactate dehydrogenase (LDH) release, the protein expression of mitogen-activated protein kinase (MAPK) pathway, and the relative expression of key genes related to apoptosis were evaluated. The results indicated that DON significantly inhibited cellular viability and promoted the release of LDH by damaging the membrane integrity of PHNCs, however, the cellular viability was increased and LDH leakage rate were decreased after adding MAPK inhibitors. DON induced PHNCs apoptosis and phosphorylation of MAPK pathway proteins dose-dependently. The ratios of phospho p-JNK/JNK and p-p38/p38 significantly increased with the increase of DON concentration, while the p-ERK/ERK ratio significantly decreased. In addition, DON upregulated the BAX mRNA level, and downregulated the BCL2 mRNA level. Pre-incubation with inhibitors of JNK (SP600125) and p38 (SB202190) significantly decreases the BAX/BCL2 ratio. However, pre-incubation with the inhibitor of ERK (U0126), significantly increased the BAX/BCL2 ratio. These data demonstrated that DON induces toxic effects and apoptosis in PHNCs via the MAPK signaling pathway.

Effects of deoxynivalenol exposure on cerebral lipid peroxidation, neurotransmitter and calcium homeostasis of chicks in vivo

&NA; During current research, the effects of deoxynivalenol (DON) exposure on cerebral lipid peroxidation, neurotransmitter secretion and calcium homeostasis in chicks were evaluated. One hundred and twenty Hailan chicks (male, 1‐day‐old) were randomly divided into four groups. Chicks in low, medium and high dose groups were fed with 0.27, 1.68 and 12.21 mg/kg−1 DON respectively by gavage according to feed intake. Chicks in control group were fed with physiological saline by gavage. The trials were conducted for 36 d. At the end of the trials, twenty chicks per group were sacrificed, and the cerebra were collected for measuring the brain indices. Compared with the control group, the activities of total superoxide dismutase (T‐SOD) and glutathione peroxidase were significantly decreased in treatment groups (P < 0.05), the contents of malondialdehyde in high dose group were increased (P < 0.05), the catalase activities and nitric oxide contents in medium and high dose groups were decreased (P < 0.05), and the activities of T‐AOC in high dose group were reduced (P < 0.05). Compared with the control group, the concentrations of norepinephrine and 5‐hydroxytryptamine in high dose group were obviously increased (P < 0.05), while the concentrations of dopamine were decreased (P < 0.05). Meanwhile, the concentrations of calcium and calmodulin (CaM) in medium and high dose groups were lower than those of the control group (P < 0.05), and the gene relative expression of CaM mRNA in treatment groups were significantly reduced (P < 0.05), in a dose‐dependent manner. These results suggested that DON exposure can affect the cerebral lipid peroxidation, neurotransmitters secretion and the balance of calcium homeostasis in chicks. HighlightsDeoxynivalenol has neurotoxic effect to the chicks.Deoxynivalenol affect lipid peroxidation of cerebrum tissue in chicks.Deoxynivalenol altere neurotransmitter levels of cerebrum tissue in chicks.Deoxynivalenol change the equilibrium of calcium homeostasis in chicks.

Therapeutic Role of Green Tea Polyphenols in Improving Fertility: A Review

Sperm cells are highly sensitive to reactive oxygen species (ROS), which are produced during cellular oxidation. In normal cell biology, ROS levels increase with a decreasing antioxidant response, resulting in oxidative stress which threatens sperm biology. Oxidative stress has numerous effects, including increased apoptosis, reduced motion parameters, and reduced sperm integrity. In this regard, green tea polyphenols (GrTPs) have been reported to possess properties that may increase the quality of male and female gametes, mostly via the capability of catechins to reduce ROS production. GrTPs have antioxidant properties that improve major semen parameters, such as sperm concentration, motility, morphology, DNA damage, fertility rate, and gamete quality. These unique properties of green tea catechins could improve reproductive health and represent an important study area. This exploratory review discusses the therapeutic effects of GrTPs against infertility, their possible mechanisms of action, and recommended supportive therapy for improving fertility in humans and in animals.