OPG/RANKL/RANK系统的生理功能

骨是一个代谢活跃的器官,在整个生命周期中都在进行不断的更新与重建。在骨形态发生和重建的过程中主要受到细胞因子和激素的调节,同时骨代谢也受到OPG/RANKL/RANK系统的调控。论文主要对骨骼生理结构与骨组织细胞之间的关系,骨代谢和骨的形成过程,OPG/RANKL/RANK系统的生理功能,OPG/RANKL/RANK系统的影响因子以及骨代谢紊乱所引起的疾病几个方面作一综述。     

Effects of RANKL, osteoprotegerin, calcium and phosphorus on survival and activation of Muscovy duck osteoclasts in vitro

The aim of this study was to determine whether receptor activator of nuclear factor NF-κB ligand (RANKL), osteoprotegerin (OPG) and a calcium:phosphorus (Ca:P) ratio of 2:1 could affect survival and activation of Muscovy duck osteoclasts (OCs). Bone marrow cells were obtained from 5-day-old Muscovy ducks and cultured with (Group A) No added factors, (B) 30 ng/mL soluble RANKL (sRANKL), (C) 30 ng/mL sRANKL and 10 ng/mL OPG, (D) 10 ng/mL OPG, (E) 50 ng/mL OPG, (F) 100 ng/mL OPG and (G) 30 ng/mL sRANKL, 6 mmol/L Ca and 3 mmol/L P.sRANKL promoted the survival of OCs on day 2, whereas the number of OCs decreased with addition of OPG in a dose-dependent manner. OPG and Ca:P (2:1) both inhibited OC survival induced by RANKL. RANKL stimulated bone resorption by OCs, whereas OPG, but not Ca:P (2:1), inhibited the activity of OCs induced by RANKL. RANKL promotes the survival and activation of OCs from Muscovy ducks, whereas OPG and, to a lesser extent, Ca:P (2:1) reduce the life span and inhibited the activation of OCs induced by RANKL.     

Effect of dietary calcium level and source on mineral utilisation by piglets fed diets containing exogenous phytase

Calcium and phosphorus are essential minerals, closely linked in digestive processes and metabolism. With widespread use of low P diets containing exogenous phytase, the optimal dietary Ca level was verified. The 40‐day study evaluated the effects of Ca level (4, 7 and 10 g/kg diet) and Ca source (Ca from CaCO₃ and from Lithothamnium calcareum) on mineral utilisation in 72 piglets (7.9 ± 1.0 kg BW) fed an exogenous phytase containing diet with 2.9 g digestible P/kg. Measured parameters were growth performance, stomach mineral solubility, bone breaking strength and urinary, serum and bone mineral concentration. The apparent total tract digestibility of minerals was also assessed in the two diets with 7 g Ca/kg, using 12 additional pigs. Regardless of Ca source, increasing dietary Ca impaired feed conversion ratio, increased urinary pH, increased serum and urinary Ca, decreased serum and urinary P, decreased serum Mg and increased urinary Mg, increased serum AP activity, decreased bone Mg increased bone Zn. Bone breaking strength was improved with 7 compared to 4 g Ca/kg. Compared to CaCO₃, Ca from Lithothamnium calcareum increased serum Mg and with, 10 g Ca/kg, it limited body weight gain. The dose response of Ca in a diet with 2.9 g digestible P/kg and including exogenous phytase indicated that: (i) a low dietary Ca was beneficial for piglet growth, but was limiting the metabolic use of P; (ii) a high dietary Ca level impaired P utilisation; (iii) the optimal P utilisation and bone breaking strength was obtained with a dietary Ca‐to‐digestible P ratio of 2.1 to 2.4:1; (iv). Increasing dietary Ca reduced Mg utilisation, but not Zn status, when fed at adequate level. Finally, Ca from Lithothamnium calcareum had similar effects on Ca and P metabolism as CaCO₃, but impaired growth when fed at the highest inclusion level.     

Expression of receptor activator of nuclear factor kappa-B ligand (RANKL) in neoplasms of dogs and cats

BACKGROUND: Receptor activator of nuclear factor kappa-B (RANK), RANK-ligand (RANKL), and the soluble decoy receptor osteoprotegerin (OPG) form a key axis modulating osteoclastogenesis. In health, RANKL-expressing bone stromal cells and osteoblasts activate osteoclasts through RANK ligation, resulting in homeostatic bone resorption. Skeletal tumors of dogs and cats, whether primary or metastatic, may express RANKL and directly induce malignant osteolysis. HYPOTHESIS: Bone malignancies of dogs and cats may express RANKL, thereby contributing to pathologic bone resorption and pain. Furthermore, relative RANKL expression in bone tumors may correlate with radiographic characteristics of bone pathology. ANIMALS: Forty-two dogs and 6 cats with spontaneously-occurring tumors involving bones or soft tissues were evaluated. METHODS: A polyclonal anti-human RANKL antibody was validated for use in canine and feline cells by flow cytometry and immunocytochemistry. Fifty cytologic specimens were collected from bone and soft tissue tumors of 48 tumor-bearing animals and assessed for RANKL expression. In 15 canine osteosarcoma (OSA) samples, relative RANKL expression was correlated with radiographic characteristics of bone pathology. RESULTS: Expression of RANKL by neoplastic cells was identified in 32/44 canine and 5/6 feline tumor samples. In 15 dogs with OSA, relative RANKL expression did not correlate with either radiographic osteolysis or bone mineral density as assessed by dual energy x-ray absorptiometry. CONCLUSIONS AND CLINICAL IMPORTANCE: In dogs and cats, tumors classically involving bone and causing pain, often may express RANKL. Confirming RANKL expression in tumors is a necessary step toward the rational institution of novel therapies targeting malignant osteolysis via RANKL antagonism.     

Inhibitory effects of osteoprotegerin on osteoclast formation and function under serum-free conditions

The purpose of this study was to determine whether osteoprotegerin (OPG) could affect osteoclat differentiation and activation under serum-free conditions. Both duck embryo bone marrow cells and RAW264.7 cells were incubated with macrophage colony stimulatory factor (M-CSF) and receptor activator for nuclear factor κB ligand (RANKL) in serum-free medium to promote osteoclastogenesis. During cultivation, 0, 10, 20, 50, and 100 ng/mL OPG were added to various groups of cells. Osteoclast differentiation and activation were monitored via tartrate-resistant acid phosphatase (TRAP) staining, filamentous-actin rings analysis, and a bone resorption assay. Furthermore, the expression osteoclast-related genes, such as TRAP and receptor activator for nuclear factor κB (RANK), that was influenced by OPG in RAW264.7 cells was examined using real-time polymerase chain reaction. In summary, findings from the present study suggested that M-CSF with RANKL can promote osteoclast differentiation and activation, and enhance the expression of TRAP and RANK mRNA in osteoclasts. In contrast, OPG inhibited these activities under serum-free conditions.     

Effects of dietary dihydroartemisinin supplementation on growth performance,hepatic inflammation,and lipid metabolism in weaned piglets with intrauterine growth retardation

The aims of this study were to investigate the effects of dietary supplementation with dihydroartemisinin (DHA) on growth performance, hepatic inflammation, and lipid metabolism in intrauterine growth retardation (IUGR)‐affected weaned piglets. Eight piglets with normal birth weight (NBW) and 16 IUGR‐affected piglets were selected and fed either a basal diet (NBW and IUGR groups) or the basal diet supplemented with 80 mg/kg DHA (IUGR‐DHA group) from 21 to 49 day of age. Blood and liver samples were collected on day 49. DHA supplementation significantly alleviated the compromised growth performance and liver damage in IUGR‐affected piglets. Additionally, DHA supplementation decreased the activities of alanine aminotransferase and aspartate aminotransferase, as well as the serum levels of non‐esterified fatty acids (NEFA), very‐low‐density lipoprotein, and total cholesterol. In the liver, the concentrations of interleukin 1 beta, interleukin 6, tumor necrosis factor alpha, triglycerides, and NEFA were decreased. Fatty acid synthesis was decreased by DHA supplementation, whereas the activities of lipoprotein lipase, hepatic lipase, and total lipase were increased. Dietary DHA supplementation led to upregulation of the expression of AMPK/SIRT1 signaling pathway‐related genes, whereas that of inflammatory factor‐related genes were downregulated. In conclusion, dietary inclusion of 80 mg/kg DHA can alleviate IUGR‐induced impairments in piglets.     

Effects of dietary methionine supplementation on growth performance,intestinal morphology,antioxidant capacity and immune function in intra‐uterine growth‐retarded suckling piglets

This study investigated the effects of dietary supplementation with _L‐methionine (_L‐Met), _DL‐methionine (_DL‐Met) and calcium salt of the methionine hydroxyl analog (MHA‐Ca) on growth performance, intestinal morphology, antioxidant capacity and immune function in intra‐uterine growth‐retarded (IUGR) suckling piglets. Six normal birthweight (NBW) female piglets and 24 same‐sex IUGR piglets were selected at birth. Piglets were fed nutrient adequate basal diet supplemented with 0.08% _L‐alanine (NBW‐CON), 0.08% _L‐alanine (IUGR‐CON), 0.12% _L‐Met (IUGR‐LM), 0.12% _DL‐Met (IUGR‐DLM) and 0.16% MHA‐Ca (IUGR‐MHA‐Ca) from 7 to 21 days of age respectively (n = 6). The results indicated that IUGR decreased average daily milk (dry matter) intake and average daily gain and increased feed conversion ratio of suckling piglets (p < 0.05). Compared with the NBW‐CON piglets, IUGR also impaired villus morphology and reduced antioxidant capacity and immune homeostasis in the intestine of IUGR‐CON piglets (p < 0.05). Supplementation with _L‐Met enhanced jejunal villus height (VH) and villus area and ileal VH of IUGR piglets compared with IUGR‐CON piglets (p < 0.05). Similarly, _DL‐Met supplementation increased VH and the ratio of VH to crypt depth in the jejunum compared with IUGR‐CON pigs (p < 0.05). Supplementation with _L‐Met and _DL‐Met (0.12%) tended to increase reduced glutathione content and reduced glutathione: oxidized glutathione ratio and decrease protein carbonyl concentration in the jejunum of piglets when compared with the IUGR‐CON group (p < 0.10). However, supplementation with MHA‐Ca had no effect on the intestinal redox status of IUGR piglets (p > 0.10). In conclusion, supplementation with either _L‐Met or _DL‐Met has a beneficial effect on the intestinal morphology and antioxidant capacity of IUGR suckling piglets.     

Immunohistochemical Localization of RANK, RANKL and OPG in Healthy and Arthritic Canine Elbow Joints

Objective— To determine if the receptor activator of nuclear factor-κB–receptor activator of nuclear factor-κB ligand–osteoprotegerin (RANK–RANKL–OPG) system is active in bone remodeling in dogs and, if so, whether differences in expression of these mediators occur in healthy and arthritic joints.
Study Design— Experimental study.
Sample Population— Fragmented processus coronoidei (n=20) were surgically removed from dogs with elbow arthritis and 5 corresponding healthy samples from dogs euthanatized for reasons other than elbow joint disease.
Methods— Bright-field immunohistochemistry and high-resolution fluorescence microscopy were used to investigate the distribution of RANK, RANKL, and OPG in healthy and arthritic joints.
Results— All 3 molecules were identified by immunostaining of canine bone tissue. In elbow dysplasia, the number of RANK-positive osteoclasts was increased. In their vicinity, cells expressing RANKL, a mediator of osteoclast activation, were abundant whereas the number of osteoblasts having the potential to limit osteoclastogenesis and bone resorption via OPG was few.
Conclusions— The RANK–RANKL–OPG system is active in bone remodeling in dogs. In elbow dysplasia, a surplus of molecules promoting osteoclastogenesis was evident and is indicative of an imbalance between the mediators regulating bone resorption and bone formation. Both OPG and neutralizing antibodies against RANKL have the potential to counterbalance bone resorption.
Clinical Relevance— Therapeutic use of neutralizing antibodies against RANKL to inhibit osteoclast activation warrants further investigation.     

中草药复方防治肉鸡骨病的研究

为探明中草药复方对肉鸡骨病的防治效果，本研究选用160只1日龄健康科宝肉鸡，随机均分为8组，分别饲养至30和60日龄，由当归、枸杞、山药等8味中草药组成复方制剂，将中草药复方以高（0.6%）、中（0.4%）、低（0.2%）3个剂量组进行治疗试验，同时设置对照组饲喂基础饲粮，分别于30、60日龄时称重，采用生化分析仪检测骨代谢相关血液生化指标维生素D受体（VDR）、骨钙素（OT）、骨保护素（OPG）、碱性磷酸酶（ALP），通过万能试验机进行三点弯曲试验，以弹性模量、屈服强度和最大应力为指标检测胫骨的骨强度，运用实时荧光定量PCR方法检测胸椎、腿软骨组织中骨保护素（OPG）、骨形态发生蛋白-2（BMP-2）、成骨细胞转录因子2（Runx-2）、细胞核因子κB受体活化因子（RANK）、细胞核因子κB受体活化因子配体（RANKL）、肿瘤坏死因子α（TNF-α）基因mRNA表达量的差异。结果显示，与对照组相比添加复方中草药组试验鸡后体重略增加；60日龄时血清中VDR、OT、OPG、ALP含量显著或极显著提高（P<0.05；P<0.01）；肉鸡的胫骨骨强度指标（弹性模量、屈服强度和最大应力）显著增加（P<0.05），且弹性模量、屈服强度、最大应力3个指标均表现为30日龄高于60日龄，与血液生化指标相符；复方中草药可显著促进OPG、BMP-2、Runx-2基因表达（P<0.05），显著抑制TNF-α、RANK、RANKL基因表达（P<0.05）。综合试验结果，兼顾活重和骨强度相关指标，复方中草药高剂量组是改善科宝肉鸡骨骼特性最优配方，且中草药复方通过提高血清中VDR、OT、OPG、ALP含量，调控OPG/RANK/RANKL系统维持骨吸收和骨形成两者动态平衡，促进骨形成的重要通路BMP-2/Smads/Runx-2中BMP-2和Runx-2基因的表达，同时抑制TNF-α基因表达，以达到改善科宝肉鸡骨骼特性，最终有效防治肉鸡的骨病。     

Bone development of suckling piglets after prenatal,neonatal or perinatal treatment with dexamethasone

In mammals, the release from growth‐inhibiting conditions results in catch‐up growth. To investigate animal evidence for whether prenatal dexamethasone (DEX) treatment leads to the development of growth restriction especially reduced mineralization of skeleton, and release from it leads to the phenomenon of catch‐up, piglets were prenatally exposed to DEX (3.0 mg/sow per day^?2) during the last 24 days of prenatal life and tested further in two different ways: discontinued at birth and continued administration of DEX (0.5 mg/kg day^?2) to piglets through 30 days of neonatal life. Using dual energy X‐ray absorptiometry methods, bone mineral density (BMD) and bone mineral content (BMC) were measured. The three‐point bending test was applied to determine the mechanical properties of the bones. Furthermore, geometric properties of the bones were assessed. Serum concentration of osteocalcin (OC) was determined. Histomorphological analysis of the ribs was also performed. The consequences of neonate DEX treatment and in utero DEX exposure were reflected in a dramatic decrease of BMD, BMC and blood serum OC concentration and geometric parameters of piglets’ bones. Prenatal action of DEX during the last 24 days of pregnancy resulted in continued neonatal modification of bone tissues, thus diminishing bone quality, and negatively influenced structural development and mechanical properties, finally increasing the risk of fractures of ribs and limb bones. Prenatal DEX treatment limited to the last 24 days of foetal life did not reduce the term birth weight and the growth of suckling piglets followed up to 30 days of neonatal life, and catch‐up in bone mineralization did not occur.     

Effect of different levels of mannan‐oligosaccharide supplementation on the apparent ileal digestibility of nutrients,N‐balance and growth performance of weaned piglets

Several studies report that dietary mannan‐oligosaccharides (MOSs) improve the growth performance of piglets, however, only a few studies focus on nutrient digestibility. The aim of the study was to investigate the effect of dietary MOS on ileal digestibility of nutrients and on N‐balance and growth performance of piglets weaned at 28 days of age. Three experiments were conducted: a digestibility trial with a total of 30, simple T‐cannulated piglets (Exp. 1), a N‐balance trial with a total of 48 intact piglets (Exp. 2) and a performance trial with a total of 324 piglets (Exp. 3). In Exp. 1 and 2, the same five dietary treatments were carried out by supplementing the basal diet with 0, 1, 2, 4 g MOS or with 0.2 g antibiotic growth promoter (AGP, Avilamycin) per kg of diet. In Exp. 3, three dietary treatments were used as follows: the basal diet was supplemented no additive, 2 g MOS or 0.2 g AGP (Avilamycin) per kg of diet. Dietary MOS (2 or 4 g/kg) enhanced the ileal digestibility of crude protein similar to antibiotics. Addition of 1 g/kg MOS significantly increased the digestibility of Ca and P by 8.4% and 7.7% units, respectively; however, further increment did not enhance the absorption. Addition of 1 or 2 g/kg MOS significantly increased the ileal digestibility of lysine, methionine, cystine and threonine with the same magnitude or even more than the AGP. In our study, MOS supplementation had no influence on N‐balance and growth parameters. Because of the better apparent ileal digestibility of certain nutrients, however, the protein, lysine, methionine, cystine, threonine, Ca and P contents of the diet can probably be reduced without weakening the rearing performance of piglets, when the diet is supplemented with 2 g/kg MOS.     

Effects of dietary calcium, available phosphorus and vitamin D on growth rate, food utilisation, plasma and bone constituents and calcium and phosphorus retention of commercial broiler strains

1. The effects of different dietary concentrations of calcium (Ca), available phosphorus (AP) and vitamin D (D) on 5- to 16-day growth performance, and aspects of calcium and phosphorus (P) metabolism of chickens from three commercial strains were studied in two experiments. 2. Increasing dietary Ca reduced weight gain, tibia Ca and P content and increased plasma total Ca, Ca consumption and excretion, whilst dietary Ca at 32 g/kg increased tibia Ca:P ratio, plasma ionized calcium and reduced plasma P, tibia ash, P excretion, excreta moisture and Ca retention. 3. Increasing dietary AP reduced plasma total and ionized Ca and excreta moisture and increased P consumption and excretion, plasma P and tibia ash. 4. The addition of vitamin D increased plasma total and ionized Ca, tibia Ca:P ratio and reduced plasma sodium and P concentrations. 5. Strains differed in their tibia contents of Ca and Ca:P ratios, in response to Ca, AP and vitamin D diets whilst they differed in Ca excretion and excreta moisture caused by feeding either dietary Ca or AP. 6. It was concluded that dietary Ca, AP, vitamin D and strain of broiler chickens influenced the metabolism of Ca and P and that, as a consequence, the tolerance to high dietary Ca. A lean strain of chickens tolerated high dietary calcium better than its fat counterparts.     

Effects of Piper sarmentosum extract on the growth performance,antioxidant capability and immune response in weaned piglets

The biological properties of Piper sarmentosum render it a potential substitute for antibiotics in livestock feed. This study evaluated the effects of P. sarmentosum extract (PSE) on the growth performance, antioxidant capability and immune response of weaned piglets. Eighty 21‐d‐old weaned piglets were selected and randomly allocated to one of four dietary treatments with five replicates of four pigs each. The dietary treatments consisted of a basal diet supplemented with 0 (T0), 50 (T50), 100 (T100) or 200 (T200) mg/kg PSE. The feeding trial lasted 4 weeks. The results revealed that the T50 group had the highest average daily gain (ADG) and average daily feed intake (ADFI) throughout the feeding trial (p < 0.05). Additionally, the T50 group had higher (p < 0.05) serum glutathione peroxidase activity (GSH‐Px) and lower (p < 0.05) serum malondialdehyde (MDA) levels than the T0 group at 4 weeks post‐weaning (p < 0.05). Serum levels of interleukin‐1β (IL‐1β) and tumour necrosis factor‐α (TNF‐α) decreased, while serum levels of interleukin‐4 (IL‐4), interleukin‐10 (IL‐10) and transforming growth factor‐β (TGF‐β) increased by PSE supplementation at 4 weeks post‐weaning (p < 0.05). PSE supplementation upregulated the mRNA expression of IL‐4, IL‐10 and TGF‐β and downregulated the mRNA expression of TNF‐α, IL‐1β and interleukin‐6 (IL‐6) in the ileal mucosal layer of piglets (p < 0.05). In summary, our study findings revealed that PSE supplementation improved the antioxidant capability, and reduced inflammation, which may be beneficial to weaned piglet health.     

Influence of l‐carnitine and l‐arginine on protein synthesis and maturation of the semitendinosus muscle of lightweight piglets

Lightweight (LW ) piglets from large litters display impaired growth performance compared with heavier littermates. This study investigated the growth performance and muscle development of early‐weaned LW piglets (birthweight <1.2 kg) from large litters (17.3 ± 3.0 total born per litter), fed ad libitum a milk replacer supplemented with either l ‐carnitine (CAR ) or l ‐arginine (ARG ) from day 7 to day 28 of age. In total, 36 female and entire male Swiss Large White piglets, weaned on day 7 of age, were artificially reared in pairs in rescue decks. They were allocated to one of three dietary treatments: unsupplemented control (CON ), 0.48 g l ‐carnitine·piglet^?1·day^?1 (CAR ) or 1.20 g l ‐arginine·kg body weight^?1·day^?1 (ARG ). Milk replacer was prepared daily in a 1:4 powder‐to‐water ratio and fed ad libitum. Piglets were weighed at birth and on days 7, 14, 21 and 28. Feed intake was assessed daily. Piglets were euthanized on day 28. The entire semitendinosus muscle (STM ) was collected, and organs were weighed. Subsequently, the STM was divided into the light (STM _l) and dark (STM _d) portion, and contractile and metabolic traits were analysed by ATP histochemistry, enzyme activities and gene expression. No differences in growth performance, organ and STM weight and on contractile traits were found between groups. A tendency (p  <  .10) for an elevated lipid oxidation enzyme activity in the STM _l and STM _d and greater (p  <  .05) phosphorylation of the mammalian target of rapamycin pathway in the STM _l of CAR compared with CON piglets was found. Despite these metabolic responses, the lack of effect of CAR and ARG supplementation on growth performance suggests that providing the milk replacer ad libitum in combination with added CAR and ARG is insufficient for eliciting faster growth of LW piglets.     

Effect of porcine glucagon‐like peptides‐2 on tight junction in GLP‐2R + IPEC‐J2 cell through the PI3k/Akt/mTOR/p70S6K signalling pathway

Because of rare glucagon‐like peptide‐2 (GLP‐2) receptor (+) cells within the gut mucosa, the molecular mechanisms transducing the diverse actions of GLP‐2 remain largely obscure. This research identified the naturally occurring intestinal cell lines that endogenously express GLP‐2R and determined the molecular mechanisms of the protective effects of GLP‐2‐mediated tight junctions (TJ) in GLP‐2R (+) cell line. (i) Immunohistochemistry results showed that GLP‐2R is localised to the epithelia, laminae propriae and muscle layers of the small and large bowels of newborn piglets. (ii) GLP‐2R expression was apparent in the cytoplasm of endocrine cells in IPEC‐J2 cell lines. (iii) The protein expressions of ZO‐1, claudin‐1, occludin, p‐PI₃K, p‐Akt, p‐mTOR and p‐p70^S6K significantly (p < 0.05) increased in GLP‐2‐treated IPEC‐J2 cells, and all of them significantly (p < 0.05) decreased when LY‐294002 or rapamycin was added. GLP‐2 improves intestinal TJ expression of GLP‐2R (+) cells through the PI₃k/Akt/mTOR/p70^S6K signalling pathway.     

Effect of ginseng polysaccharides on the immunity and growth of piglets by dietary supplementation during late pregnancy and lactating sows

There are many reports that dietary supplementation with plant polysaccharides in pigs might promote their growth, but little is known about the maternal effect of ginseng polysaccharides (GPS) on piglets’ growth by dietary supplementation to pregnant and lactating sows. In the current study, the effects of dietary supplementation with GPS on the immunity of sows and growth of their piglets were investigated. Results showed no significant difference among the four groups in the total number of piglets, live piglets, weak piglets and birth weight of piglets, indicating the GPS‐treatment has no adverse effect on reproduction. Furthermore, the weaning weight of the GPS‐treated groups was higher than that of control group (P < 0.05); among them, the addition of 200 mg/kg dose has the best effect. Interestingly, GPS increased the total immunoglobulin G concentration in milk and serum of sows (P < 0.05). The concentrations of interleukin (IL)‐2, IL‐6, tumor necrosis factor (TNF)‐α, and interferon‐γ in milk and serum of sows were also increased in the experimental groups relative to the control (P < 0.05). Meanwhile, maternal supplementation of GPS significantly increased IL‐2 and TNF‐α concentration in the piglets’ serum of the experimental groups relative to control (P < 0.05). GPS (200 mg/kg) significantly increased the glutathione peroxidase activity in milk and serum (P < 0.05), while the concentrations of malondialdehyde were significantly reduced (P < 0.05). The present results indicated that GPS supplementation during late pregnancy and lactation improved immunity‐related bio‐molecular levels in sow serum and milk, which may be further beneficial to piglet health and growth through biological transmission effects.     

Involvement of the Ca2+ signaling pathway in osteoprotegerin inhibition of osteoclast differentiation and maturation

The purpose of this study was to determine whether the Ca²⁺ signaling pathway is involved in the ability of osteoprotegerin (OPG) to inhibit osteoclast differentiation and maturation. RAW264.7 cells were incubated with macrophage colony-stimulating factor (M-CSF) + receptor activator of nuclear factor-κB ligand (RANKL) to stimulate osteoclastogenesis and then treated with different concentrations of OPG, an inhibitor of osteoclast differentiation. The intracellular Ca²⁺ concentration [Ca²⁺]_i and phosphorylation of Ca²⁺/calmodulin-dependent protein kinase II (CaMKII) in the different treatment groups were measured by flow cytometry and Western blotting, respectively. The results confirmed that M-CSF + RANKL significantly increased [Ca²⁺]_i and CaMKII phosphorylation in osteoclasts (p < 0.01), and that these effects were subsequently decreased by OPG treatment. Exposure to specific inhibitors of the Ca²⁺ signaling pathway revealed that these changes varied between the different OPG treatment groups. Findings from the present study indicated that the Ca²⁺ signaling pathway is involved in both the regulation of osteoclastogenesis as well as inhibition of osteoclast differentiation and activation by OPG.     

Water extract of Artemisia ordosica enhances antioxidant capability and immune response without affecting growth performance in weanling piglets

The present experiment was conducted to investigate the effects of water extract of Artemisia ordosica (WEAO) on growth performance, antioxidant capability and immune response in weanling piglets. Seventy‐two 28‐day‐old weanling piglets were randomly allocated into four treatments with six replicate pens per treatment and three piglets per pen (n = 18). These four treatment diets were formulated by adding 0, 250, 500 and 750 mg/kg WEAO to the basal diet. The experiment lasted for 28 days. Body weight and feed intake were measured. Blood samples were collected to determine immune and antioxidative parameters. The experimental results showed that WEAO supplementation improved the apparent nutrient digestibility of piglets in a linear or quadratic dose‐dependent manner. In addition, dietary WEAO quadratically increased serum concentrations of cytokines interleukin (IL)‐1, IL‐4, tumour necrosis factor (TNF)‐α, soluble surface antigen CD8 (sCD8), immunoglobulins (Ig)‐A and linearly increased serum concentrations of IL‐2, IL‐6, IgG, IgM. Furthermore, dietary WEAO linearly or quadratically decreased serum concentrations of malondialdehyde but quadratically increased activities of antioxidant enzymes and total antioxidative capacity. These results suggested that WEAO may prove useful as a natural phytogenic feed additive with antioxidative potential and could be incorporated into diets of piglets.     

Tolerance evaluation of overdosed dietary levels of 25‐hydroxyvitamin D3 in growing piglets

Forty‐eight, cross‐bred (GL × LW × P) piglets were used in a 42‐day tolerance trial to assess the effects of feeding diets supplemented with vitamin D or increasing levels of 25‐hydroxyvitamin D₃ (25‐OH‐D₃). Six‐week‐old piglets (24 castrate males, 24 females) were used. Two replicate groups of 6 piglets were randomized by weight and allocated to four dietary treatments. The control group (T1) was supplemented with 50 μg vitamin D₃/kg feed. The experimental groups received 25‐OH‐D₃ at the recommended dose (T2: 50 μg/kg = 1x), at 250 μg/kg (T3: 5x) or at 500 μg/kg (T4: 10x) respectively. Feed intake and daily weight gain were measured weekly, and the animals were examined by a veterinarian daily. After 42 days, body mass, blood, urine, bone and tissue samples were analysed and a pathology examination conducted. Dietary treatments had no significant effect on final body mass or daily weight gain. The 25‐OH‐D₃ plasma concentration in T1 was 17 ± 3 ng/ml (mean ± SD) while the respective values of the experimental groups were significantly increased in T2, T3 and T4. Tissue concentrations of 25‐OH‐D₃ were higher in liver and muscle for T3 and T4 and in skin for T4 than in T1. However, neither gross pathology nor histology, nor blood and urine characteristics, nor bone parameters were affected by dietary treatments. Weight of organs as well as dry matter, ash and calcium content of kidneys remained unaffected by dietary 25‐OH‐D₃ intake. Furthermore, no changes were observed for general indicators of health. The results of this study demonstrated that feeding piglets with 25‐OH‐D₃ at 5 or 10 times the recommended level had no adverse effects on any of the biological parameters measured. It was concluded that 25‐OH‐D₃ can be regarded as a supplement with a very high safety margin when used at the recommended level.     

Body weight of young broilers fed with declining calcium and phosphorus contents during the starter period is irresponsive to changes in the skeleton

To evaluate the effect of reduced calcium (Ca) and available P (aP) levels in starter diets on growth performance and tibia, sternum and serum characteristics, and to assay the association between Ca and P (aP) intakes and these variables, 600 one-day-old broiler chicks (Ross 308) were used in a 21-days trial. Broilers were randomly allotted to five treatments with five replicate pens. Chicks were fed on one of the five starter diets that were formulated with a 10% reduction in the Ca and aP contents, starting from the control diet and hence, named as C (0.96% Ca, 0.45% aP), L1 (0.85% Ca and 0.42% aP), L2 (0.77% Ca and 0.38% aP), L3 (0.68% Ca and 0.34% aP) and L4 (0.61% Ca and 0.31% aP). Declining dietary Ca and aP levels did not affect body weight and levels of Ca, P and alkaline phosphatase in serum, but reduced linearly feed intake, the feed conversion ratio and the bone weight and ash content. The Ca and aP intakes and Ca:aP ratio displayed a positive correlation with both tibia and sternum weights, and the mass of the bones were equally sensitive to dietary Ca and aP levels. In conclusion, the body weight of young broilers was irresponsive to changes in the skeleton when exposed to early dietary Ca and aP restrictions (up to 0.61% Ca and 0.31% aP) and the mechanostat had limits due to the association between Ca and aP intakes, and bone properties.