Selenylation modification: enhancement of the antioxidant activity of a Glycyrrhiza uralensis polysaccharide.

Selenium (Se) is an essential trace element for human beings and many other forms of life. Organic selenium from natural foods has greater bioavailability and is safer than inorganic selenium species. In this article, the structural properties and antioxidant activities of a Glycyrrhiza uralensis polysaccharide (GUP) after selenylation modification were investigated. The GUP was extracted by water decoction and ethanol precipitation and purified via protein elimination using the trichloroacetic acid method and column chromatography. The purified product was subsequently modified by the nitric acid-sodium selenite (HNO3-Na2SeO3) method. The selenized GUP (SeGUP) product was characterized by Fourier transform-infrared (FT-IR) spectroscopy, and its thermal stability, particle size, and antioxidant activities were investigated. FT-IR analysis indicated that the selenium in SeGUP existed mainly as O-Se-O. The thermal stability and particle size of SeGUP differed significantly from those of GUP. Moreover, compared to GUP, SeGUP exhibited greater antioxidant activities in vitro and in vivo. These results indicate that selenylation modification significantly enhances the antioxidant activity of SeGUP, increasing its potential for application as an antioxidant. Graphical abstract ᅟ.

[Exploration and characterization of a universal piggyBac transposon vector for efficient transgene studies in sheep].

piggyBac (PB) is a DNA transposon that mediates transposition in various animal cells. As an efficient tool, PB transposons are applied to various transgenic research and optimized for different species, which is an essential means to enhance its versatility. To construct a universal PB transposase (PBase) vector for ovine transgene manipulation, the gene expression box of PBase with optimized bias for sheep codons was inserted into the pBNW-TP1 vector. The vector pBNW-TP2 with a single plasmid was successfully constructed. Then, pBNW-TP2 was transfected into ovine fibroblasts and mammary epithelial cells. The stable transfected cell lines were selected by G418 and the PB transposition sites were determined by Tail-PCR in the stable transfected cell lines. The transposition efficiency was verified by student's t-tests for cell clones with methylene blue staining. The results showed that pBNW-TP2 successfully guided the production of transgenic ovine fibroblasts and mammary epithelial cell lines. The PB transposition test indicated that the pBNW-TP2 vector can be specifically integrated into the TTAA sites of the sheep genome. The statistical analysis of methylene blue staining results suggested that the transgenic efficiency mediated by the pBNW-TP2 vector increased significantly. In summary, we verified and analyzed characteristics of the universal PB transposon vector pBNW-TP2 for sheep in this study, which will provide a scientific basis for future transgenic research mediated by the PB transposon in ovine somatic cells.

Optimized Extraction, Preliminary Characterization, and In Vitro Antioxidant Activity of Polysaccharides from Glycyrrhiza Uralensis Fisch.

BACKGROUND This study performed optimized extraction, preliminary characterization, and in vitro antioxidant activities of polysaccharides from Glycyrrhiza uralensis Fisch. MATERIAL AND METHODS Three parameters (extraction temperature, ratio of water to raw material, and extraction time) were optimized for yields of G. uralensis polysaccharides (GUP) using response surface methodology with Box-Behnken design (BBD). The GUP was purified using DEAE cellulose 32-column chromatography. The main fraction obtained from G. uralensis Fisch was GUP-II, which was composed of rhamnose, arabinose, galactose, and glucose monosaccharide, was screened for antioxidant properties using DP Hand hydroxyl radical scavenging assays. In addition, immunological activity of GUP-II was determined by nitric oxide and lymphocyte proliferation assays. RESULTS Optimization revealed maximum GUP yields with an extraction temperature of 99°C, water: raw material ratio of 15: 1, and extraction duration of 2 h. GUP-II purified from G. uralensis Fisch had good in vitro DPPH and hydroxyl radical scavenging abilities. Immunologically, GUP-II significantly stimulated NO production in RAW 264.7 macrophages, and significantly enhanced LPS-induced lymphocyte proliferation. CONCLUSIONS Extraction of GUP from G. uralensis Fisch can be optimized with respect to temperature, extraction period, and ratio of water to material, using response surface methodology. The purified product (GUP-II) possesses excellent antioxidant and immunological activities.

The anti-inflammatory and antioxidant effects of leonurine hydrochloride after lipopolysaccharide challenge in broiler chicks.

This study was carried out to investigate the protective effects of leonurine hydrochloride (LH, from Leonurus sibiricus) on lipopolysaccharide (LPS)-stimulated broiler chicks. A total of 120 one-day-old male Ross broilers were randomly divided into 4 treatment groups with 6 replicates of 5 birds per cage. The experiment was designed as a 2 × 2 factorial arrangement with LH (0 or 120 mg/kg) and LPS (injection of saline or 1.5 mg/kg body weight) levels as treatments. On days 14, 16, 18, and 20 of the trial, broilers were intraperitoneally injected with LPS or saline. Blood, spleen, and liver samples were collected on days 21 and 28 for analysis. The results showed that dietary LH had no effect on growth performance or immunoglobulin concentrations in the serum. However, dietary LH prevented LPS-induced reductions in average daily gain and average daily feed intake in the broilers on days 15-21 of the trial (P > 0.05). Dietary LH supplementation dramatically attenuated the LPS-induced increases in the spleen index, reduced glutathione (GSH) activity (serum and liver) and total superoxide dismutase (T-SOD) activity (serum and spleen), and significantly reduced malondialdehyde (MDA) levels (serum, spleen, and liver) on days 21 and 28 (P < 0.05). Additionally, LH supplementation significantly mitigated the LPS-induced increases in the tumor necrosis factor (TNF)-α (serum and spleen), interleukin (IL)-1ß (serum, spleen and liver), IL-2 (liver), IL-6 (serum, spleen and liver), toll-like receptor 4 (TLR4) (spleen and liver), and nuclear factor (NF)-κB (spleen and liver) levels on days 21 and 28 (P < 0.05). Therefore, this study revealed that LH could downregulate the expression of proinflammatory factors, mainly by inhibiting the expression of TLR4 and the activation of NF-κB. LH may be a potential feed additive with dual efficacy as an anti-inflammatory and antioxidant agent.

Dietary leonurine hydrochloride supplementation attenuates lipopolysaccharide challenge-induced intestinal inflammation and barrier dysfunction by inhibiting the NF-κB/MAPK signaling pathway in broilers.

This study was performed to evaluate the beneficial effects of dietary leonurine hydrochloride (LH) supplementation on intestinal morphology and barrier integrity and further illuminate its underlying antioxidant and immunomodulatory mechanisms in lipopolysaccharide (LPS)-treated broilers. A total of 120 1-d-old male broilers (Ross 308) were assigned to 4 treatment groups with 6 replicates of 5 birds per cage. The experiment was designed in a 2 × 2 factorial arrangement with LH (0 or 120 mg/kg) and LPS (injection of saline or 1.5 mg/kg body weight) as treatments. On days 14, 16, 18, and 20 of the trial, broilers were intraperitoneally injected with LPS or physiological saline. Compared with the control group, LPS-challenged broilers showed impaired growth performance (P < 0.05) from day 15 to day 21 of the trial, increased serum diamine oxidase (DAO) and D-lactic acid (D-LA) levels coupled with reduced glutathione (GSH) content and total superoxide dismutase (T-SOD) activity (duodenal and jejunal mucosa), reduced malondialdehyde (MDA) content (duodenal, jejunal, and ileal mucosa), and compromised morphological structure of the duodenum and jejunum. Additionally, LPS challenge increased (P < 0.05) the mRNA expression of proinflammatory cytokine genes and reduced tight junction (TJ) protein expression in the jejunum. However, dietary LH prevented LPS-induced reductions in average daily gain (ADG) and average daily feed intake (ADFI) in broilers. It also alleviated LPS challenge-induced increases in serum DAO levels, MDA content (duodenal and jejunal mucosa), and jejunal crypt depth (P < 0.05) but reduced villus height, GSH content (jejunal mucosa), and T-SOD activity (duodenal and jejunal mucosa) (P < 0.05). Additionally, LH supplementation significantly downregulated the mRNA expression of nuclear factor (NF)-κB, cyclooxygenase-2 (COX-2), and proinflammatory cytokines (TNF-α, IL-1ß, and IL-6) and upregulated the mRNA expression of zonula occludens-1 (ZO-1) and Occludin in the jejunal mucosa induced by LPS (P < 0.05). On the other hand, LH administration prevented LPS-induced activation of the p38, extracellular signal-regulated kinase (ERK) and c-Jun N-terminal kinase (JNK) mitogen-activated protein kinases (MAPKs) and attenuated IkB alpha (IκBα) phosphorylation and nuclear translocation of NF-κB (p65) in the jejunal mucosa. In conclusion, dietary LH supplementation attenuates intestinal mucosal disruption mainly by accelerating the expression of TJ proteins and inhibiting activation of the NF-κB/MAPK signaling pathway.

Effects of berberine on the growth performance, antioxidative capacity and immune response to lipopolysaccharide challenge in broilers.

This study evaluated the effects of berberine on growth performance, immunity, haematological parameters, antioxidant capacity, and the expression of immune response-related genes in lipopolysaccharide (LPS)-challenged broilers. We assigned 120 one-day-old male broilers (Ross 308) to two treatment groups; each group included two subgroups, each of which included six replicates of five birds per replicate. The experiment used a 2 × 2 factorial arrangement with berberine treatment (0 or 60 mg/kg dietary) and challenge status [injection of saline (9 g/L w/v) or LPS (1.5 mg/kg body weight)] as the main factors. On days 14, 16, 18 and 20, broilers were intraperitoneally injected with LPS or physiological saline. Blood and liver samples were collected on day 21. Dietary berberine supplementation significantly alleviated the compromised average daily gain and average daily feed intake (p < 0.05) caused by LPS. The LPS challenge led to increased lymphocyte and white blood cell (WBC) counts, malondialdehyde (serum and liver) content, and immunoglobulin G and M, tumour necrosis factor-α (TNF-α) and interleukin-1ß (IL-1ß) expression (p < 0.05) and significantly reduced serum total superoxide dismutase (T-SOD) activity (p < 0.05). Dietary berberine significantly mitigated the LPS-induced decreases in the mRNA expression of nuclear factor-kappa B (NF-κB), TNF-α, IL-1ß, inducible nitrite synthase and cyclooxygenase-2 (p < 0.05) in the liver. In conclusion, berberine supplementation has a positive effect on LPS challenge, which may be related to the increase in antioxidant enzyme activity and inhibition of both NF-κB signalling and the expression of inflammatory mediators.

Effect of Glycyrrhiza uralensis Fisch polysaccharide on growth performance and immunologic function in mice in Ural City, Xinjiang.

OBJECTIVE: To discuss the effect of Glycyrrhiza uralensis (G. uralensis) Fisch polysaccharide on growth performance and immunologic function in mice in Ural City, Xinjiang and to provide important data supporting the application of Glycyrrhiza polysaccharide. METHODS: A total of 100 Kunming mice aged 3 weeks old were randomly divided into 5 groups with 20 mice in each group (10 were females and 10 were males). About 0.5 mL normal saline was given to the mice of control group every day and 0.5 mL G. uralensis Fisch polysaccharide was given to the mice of other groups at the concentration of 1, 20, 50 and 100 mg/mL, respectively. The growth performance (average body weight, average daily feed intake and feed efficiency), immune organ indexes (spleen index and thymus index) and immunologic function (serum IL-2, CD4+/CD8+ and the activity of NK cells) of mice in each group were detected continuously. RESULTS: The average body weight, feed efficiency, serum IL-2, CD4+/CD8+ and the activity of NK cells of mice were increased with the increase of administrated time after administrating G. uralensis Fisch polysaccharide and were reached up the largest level on Day 28. At the same time, each index was proportional to the given dose and was significantly higher than those of control group and reached up the largest level at the administrated dose of 100 mg/mL. After administrating G. uralensis Fisch polysaccharide, the spleen index and thymus index of mice were increased with the increase of administrated dose and the spleen index and thymus index of mice administrated with the dose of 100 mg/mL were maximum which was more than 1.51 times and 1.43 times of that in control group, respectively and the comparative differences showed statistical significance (P < 0.05). The average daily feed intake of mice in each group was increased with the passage of time and at the same time, the comparison of average daily feed intake of mice in each group was not significantly different (P > 0.05). CONCLUSIONS: G. uralensis Fisch polysaccharide can significantly improve the growth performance and immunologic function of mice and laid a research basis for the clinical application of G. uralensis Fisch polysaccharide.