Thyroid hormones alter estrous cyclicity and antioxidative status in the ovaries of rats

To expand our understanding of the roles of thyroid hormones on female reproduction, we induced hypo‐ and hyper‐T rat models to investigate the roles of thyroid hormones on estrous cyclicity, as well as the antioxidative status in the ovaries of rats. In the current study, our data show that hypothyroidism (hypo‐T) and hyperthyroidism (hyper‐T) led to significantly reduced body weights and ovarain weights and delayed vaginal opening day. For hyper‐T, thyroxine (T4), tri‐iodothyronine (T3), progesterone (P4) and follicle‐stimulating hormone (FSH) were significantly increased, while estradiol (E2) and luteinizing hormone (LH) were significantly decreased. For hypo‐T rats, serum levels of total T4 and T3, E2, P4, FSH and LH were significantly increased, while concentrations of E2 and LH were significantly decreased. For ovary morphology, the numbers of secondary and antral follicles were significantly decreased with more atretic antral follicles and less corpora lutea in both hyper‐ and hypo‐T groups. Both hyper‐T and hypo‐T treatment significantly decreased the expressions of thyroid hormone receptor α1 in the ovary. Hypo‐T significantly reduced nitric oxide (NO), total NO synthase (tNOS), inducible NOS and constitutive NOS activities, but hyper‐T increased them. For antioxidative parameters, hypo‐T and hyper‐T treatment significantly increased malondialdehyde (MDA) contents. The activities of both glutathione peroxidase (GSH‐Px) and catalase (CAT) significantly decreased in the hypo‐T group but increased in the hyper‐T group. Total superoxide dismutase (T‐SOD) activity was significantly increased in the hyper‐T group. In summary, thyroid hormones alter estrous cyclicity and antioxidative status in the ovary of the rat may act through the NOS signaling pathway.     

Expression and regulation of Foxa2 in the rat uterus during early pregnancy

The forkhead box a (Foxa) protein family has been found to play important roles in mammals. Recently, the expression of Foxa2 was reported in the mouse uterus, and it was reported to be involved in regulation of implantation. However, the regulation of Foxa2 expression in the uterus is still poorly understood. Therefore, the present study was conducted to investigate the expressional profiles of Foxa2 in the rat uterus during the estrus cycle and pregnancy. Furthermore, the effect of steroid hormones and Hedgehog protein on the expression of Foxa2 was analyzed in vivo and in vitro. In this study, the level of expression of Foxa2 was low in the rat uterus during the different stages of the estrus cycle. However, the expression increased transiently during early pregnancy at 3.5 days post coitus (dpc) and decreased at 5.5 dpc. In ovariectomized rats, P4 treatment had no effect on the expression of Foxa2 compared with the expression in control animals. Moreover, the expression of Foxa2 in cultured epithelial cells was not increased by P4 treatment in vitro. However, Foxa2 expression was significantly decreased in the rat uterus after 24 h of E2 treatment. Treatment of cells with a recombinant Hedgehog protein significantly increased the expression of Foxa2. These results suggest that the expression of Foxa2 may transiently increase just before the implantation and it may be regulated by E2 and Hedgehog protein.     

Effects of delayed serum separation and long-term storage on the measurement of thyroid hormones in equine blood samples

Studies were conducted to determine the effects of delaying the separation of serum from the clot and of long-term storage of serum samples on the measurement of thyroid hormones in blood from horses using a fluorescence polarization immunoassay. The measured concentrations of T3 and T4 were not affected by leaving serum on the clot for as long as 24 hours at room temperatures. Storage of serum for 19 to 22 months at -20 degrees C resulted in significant increases of measured T4, but not T3. These studies support previous work demonstrating that thyroid hormones are resistant to degradation, immunologically stable, and reasonably insensitive to potential problems of routine specimen handling when measured with an immunoassay.     

Endometrial nitric oxide production and nitric oxide synthases in the equine endometrium: Relationship with microvascular density during the estrous cycle

Nitric oxide (NO) plays an important role in angiogenesis and in the regulation of the blood flow. This study was carried out to investigate (i) the effects of endogenous estrogens and progestins and exogenous progesterone (P₄) (5 ng/ml or 1 μg/ml) or estradiol 17β (E₂β) (50 pg/ml or 1 μg/ml) on in vitro endometrial NO synthesis; (ii) the presence of different isoforms of NO synthase; (iii) and their relationship to microvascular density in the equine endometrium during the estrous cycle. NOS expression was also evaluated in the myometrium. Expression of endothelial and inducible forms of NOS in the uterus was assessed by Western blot and immunocytochemistry. Vascular density in endometrial tissue was determined on histologic sections. In the luteal phase, compared to the follicular phase, endometrial NO production increased without exogenous hormones and with exogenous E₂β (1 μg/ml). Although immunocytochemistry revealed iNOS and eNOS expression in the endometrium, no positive signal for iNOS was detected by Western blot. Endothelial NOS was observed in endometrial glands, endothelial cells, fibroblasts, blood and lymphatic vessels. Endometrial eNOS expression was the highest in the follicular and mid-luteal phases while it was found to be the lowest in the early luteal phase. In the follicular phase, hyperplasia of endometrial tissue with respect to myometrium was detected. No difference in vascular density was present between phases. All together, NO may play some roles in both proliferative and secretory phases of endometrial development in the mare.     

Effects of growth hormone secretagogues on the release of adenohypophyseal hormones in young and old healthy dogs

The effects of three growth hormone secretagogues (GHSs), ghrelin, growth hormone-releasing peptide-6 (GHRP-6), and growth hormone-releasing hormone (GHRH), on the release of adenohypophyseal hormones, growth hormone (GH), adrenocorticotropic hormone (ACTH), thyroid-stimulating hormone (TSH), luteinising hormone (LH), prolactin (PRL) and on cortisol were investigated in young and old healthy Beagle dogs. Ghrelin proved to be the most potent GHS in young dogs, whereas in old dogs GHRH administration was associated with the highest plasma GH concentrations. The mean plasma GH response after administration of ghrelin was significantly lower in the old dogs compared with the young dogs. The mean plasma GH concentration after GHRH and GHRP-6 administration was lower in the old dogs compared with the young dogs, but this difference did not reach statistical significance. In both age groups, the GHSs were specific for GH release as they did not cause significant elevations in the plasma concentrations of ACTH, cortisol, TSH, LH, and PRL. It is concluded that in young dogs, ghrelin is a more powerful stimulator of GH release than either GHRH or GHRP-6. Ageing is associated with a decrease in GH-releasing capacity of ghrelin, whereas this decline is considerably lower for GHRH or GHRP-6.     

Relationships between thyroid status, tissue oxidative metabolism, and muscle differentiation in bovine fetuses

The temporal relationships between thyroid status and differentiation of liver, heart and different skeletal muscles were examined in 42 bovine fetuses from day 110 to day 260 of development using principal component analysis of the data. Plasma concentrations of reverse-triiodothyronine (rT(3)) and thyroxine (T(4)) increased during development from day 110 to day 210 or 260, respectively, whereas concentration of triiodothyronine (T(3)) and hepatic type-1 5'-deiodinase activity (5'D1) increased from day 180 onwards. On day 260, high T(4) and rT(3) and low T(3) concentrations were observed together with a mature 5'D1 activity. Cytochrome-c oxidase (COX) activity expressed per mg protein increased at day 180 in masseter and near birth in masseter, rectus abdominis and cutaneus trunci muscles (P<0.05). Significant changes in citrate synthase (CS) activity per mg protein were observed between day 110 and day 180 in the liver and between day 210 and day 260 in the liver, the heart and the longissimus thoracis muscle (P<0.05). Muscle contractile differentiation was shown by the disappearance of the fetal myosin heavy chain from day 180 onwards. A positive correlation (r>0.47, P<0.01) was shown between thyroid status parameters (5'D1, concentrations of T(4) and T(3)) and COX activity in muscles known to be oxidative after birth (masseter, rectus abdominis) but not in liver and heart, nor in muscles known to be glycolytic after birth (cutaneus trunci, longissimus thoracis). A similar correlation was found between thyroid parameters and CS activity in liver and masseter. Results indicate that elevation of plasma T(3) concentrations in the last gestational trimester could be involved in the differentiation of oxidative skeletal muscles.     

Assessment of Uterine Vascular Perfusion During the Estrous Cycle of Mares in Connection to Circulating Leptin and Nitric Oxide Concentrations

The objective of this study was to find difference in vascular perfusion of uterine horns or uterine body throughout the estrous cycle and their relation to circulating nitric oxide and leptin concentrations. Five cyclic mares were subjected to transrectal Doppler ultrasonography and blood sampling for 18 days. Area of color and power Doppler modes was measured in pixels. Day (P = .0001) of the estrous cycle and ovulation (P = .0001) influenced uterine blood flow. Uterine body blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day 0 (day of ovulation), and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until days 12 and 10, respectively. Conversely to the contralateral uterine horn, ipsilateral uterine horn blood flow directed away from the transducer (blue, P = .0001) increased from day −5 until day −1, and its power (P = .0001) blood flow increased from day −6 until day 0; then, both decreased until day 10. Nitric oxide concentrations (P = .0001) attained two major peaks; the first on day −3 and the other persisted from day 2 until day 5. Leptin concentrations increased (P > .001) with a maximum value on day 0 and then decreased until a minimum value on day 9. In conclusion, during the estrous cycle, ipsilateral uterine horn and uterine body blood vessels had similar blood flow. Both leptin and nitric oxide played a role during follicle growth, ovulation, and corpus luteum development and modulated uterine blood flow before and after ovulation.     

Effects of branched-chain amino acids on immune status of young racing horses

High-intensity exercise and competition are associated with depressed immune function. Young horses, which participate in high-intensity exercise and competitions, are at increased risk for the development of infectious disease due to depression of immune function. The effects of branched-chain amino acid (BCAA) supplementation on the immune status of young racing horses were evaluated, determining whether BCAA might help to avoid or reduce immune suppression during exercise and competitions. Twenty horses (10 male and 10 female) were treated with BCAA supplementation; another twenty untreated horses (10 male and 10 female) constituted control group. Peripheral blood was collected from each animal and evaluated for lymphocyte subsets, phagocytosis analysis of monocytes and granulocytes, lymphocyte proliferative response, and expression of cytokine-encoding messenger ribonucleic acids (mRNAs). The numbers of CD4⁺, CD8⁺, and major histocompatibility complex (MHC) class II⁺ cells in females of the treated group were significantly higher than those in females of the control group. The lymphocyte proliferative response in female of the treated group also was significantly higher than that in females of the control group. In addition, expression of mRNAs encoding interleukin-1β (IL-1β) and interferon-γ (IFN-γ) in females of the treated group was significantly higher than that in females of the control group. There were no significant differences between males of the treated and control groups. The results of this study indicated the positive effects of BCAA supplementation in counteracting immunosuppression in young female racing horses during and following high-intensity exercise.     

The immune status of the bovine uterus during the peripartum period

The post-partum period in cattle is characterised by an increased risk of infection of the uterus, as the anatomical barriers are broached during parturition and remain open for several days. Infection of the uterus is largely influenced by the balance between bacterial contamination and the local and systemic immune status during pregnancy and around parturition. Infectious diseases are more prevalent during this period, because of an impaired immune status before and immediately after parturition. Neutrophils play a primary role in the defence of the uterus against infection. Influx of neutrophils into the uterus is thought to be mediated by chemoattractants, chemokines and adhesion molecules, such as beta2-integrin (complement receptor 3) and L-selectin (CD62L). Other cellular components activated in the uterus during this period include lymphocytes, eosinophils, mast cells and macrophages. The major classes of immunoglobulins (IgM, IgA and IgG), either by passive diffusion or local production, play an important protective role in the uterus by acting as opsonins to enhance phagocytosis, stimulating the complement pathways or blocking pathogens from adhering to mucosal surfaces. Endometrial cells express toll-like receptor 4 (TLR4), which recognises lipopolysaccharides of Escherichia coli and other Gram negative bacteria, the most common causes of bovine endometritis. Activation of TLR4 triggers the production of tumour necrosis factor alpha and other pro-inflammatory cytokines. The periparturient period is also characterised by an increased secretion of prostaglandin F(2alpha), which enhances uterine immune defences.     

不同属种肉类对大鼠血清生化指标与抗氧化能力的影响

探讨了不同属种肉类对大鼠血清生化指标和抗氧化机能的影响。试验选取体重约177 g的雄性SD大鼠90只,随机分为3组即狗肉组、羊肉组、鸭肉组,分别将日粮中的主要蛋白源替换为狗肉、羊肉、鸭肉(CP 31%)。试验期为4周。结果表明:饲喂狗肉和羊肉能显著提高大鼠生长性能,且前者使得大鼠血清碱性磷酸酶(ALP)活性极显著升高(P<0.01);而饲喂鸭肉使得大鼠血清丙氨酸转移酶(ALT)活性显著升高(P<0.05),天冬氨酸转移酶(AST)、血清尿素氮(BUN)活性极显著升高(P<0.01)。不同属种的肉之间,鸭肉组肝脏和肾脏中谷胱甘肽过氧化物酶(GSH-Px)活性均极显著升高(P<0.01)。结果提示:饲喂狗肉和羊肉能促进机体蛋白质合成,有利于蛋白质沉积;饲喂鸭肉则具有增强机体抗氧化的能力。     

Effects of saccharicterpenin on antioxidant status and urinary metabolic profile of rats

Saccharicterpenin is a new green additive agent that is derived from the extract of Theaceae plants and has the ability to improve immunity and meat quality, increase the digestive enzyme activity, and enhance the intestinal development and growth of animals. However, the antioxidant status and systematic changes in metabolic biochemistry associated with saccharicterpenin supplementation in animals are still unknown. This study examined the effects of saccharicterpenin on the antioxidant status and urinary metabolic profile of rats. Sixteen rats were randomly distributed to 2 groups. One group was treated with 400 mg/kg body weight of saccharicterpenin, and the other group was treated with equal amount of saline. Results revealed that saccharicterpenin significantly increased the capacities of anti-hydroxyl radical (13.18%) and anti-superoxide anion (14.36%), the total antioxidant capacity (48.27%), and the activities of total superoxide dismutase (3.68%), catalase (21.52%), glutathione peroxidase (5.83%) and glutathione S-transferase (29.59%) (P < 0.05). By contrast, the contents of malondialdehyde and glutathione were not significantly affected by saccharicterpenin (P > 0.05). Saccharicterpenin supplementation significantly increased the urinary levels of bile acids, ethanol, α-ketoglutarate, and α-hydroxybutyrate but decreased the level of N-acetylglutamate (P < 0.05). In summary, saccharicterpenin can enhance the antioxidant capacity and modulate the metabolism in rats.     

Effects of polyphenol‐rich plant products from grape or hop as feed supplements on the expression of inflammatory,antioxidative, cytoprotective and endoplasmic reticulum stress‐related genes and the antioxidative status in the liver of piglets

Recent studies have shown that supplementation of plant products rich in polyphenols exerts anti‐inflammatory effects in the small intestine and improves feed conversion in piglets. This study aimed to investigate whether dietary polyphenols have also anti‐inflammatory and cytoprotective effects in the liver of piglets. For this end, relative mRNA concentrations of eight genes involved in proinflammatory pathways, eight genes involved in the antioxidative and cytoprotective system, six genes of phase I and phase II metabolism and 15 genes of the unfolded protein response (triggered by stress of the endoplasmic reticulum) in the liver of pigs fed diets supplemented with either 1% of grape seed and grape marc meal extract (GME) or 1% spent hops (SH) as sources of polyphenols were determined. Relative mRNA concentrations of almost all these genes, with few exceptions, in the liver of pigs supplemented with GME or SH did not differ from those in the liver of control piglets. Gene expression data were validated by consideration of concentrations of some selected proteins of these pathways which also did not differ between piglets supplemented with GME or SH and control piglets. Moreover, concentrations of thiobarbituric acid‐reactive substances and tocopherols as well as the total antioxidant capacity in liver and plasma did not differ between pigs supplemented with either GME or SH and control piglets. Overall, this study shows that supplementation of GME or SH as sources of polyphenols does not influence hepatic pathways linked to inflammation, the antioxidant and cytoprotective system, stress of the endoplasmic reticulum and the xenobiotic system in healthy piglets.     

Effects of trimethoprim-sulfadiazine on thyroid function of horses

Trimethoprim-sulfadiazine was administered to horses in a randomized, placebo controlled study to determine the effects of potentiated sulfonamides on thyroid function in normal horses. The treatment group included eight horses that received trimethoprim-sulfadiazine mixed with molasses orally at 30 mg/kg once daily for eight weeks. The control group included 8 horses that received an oral placebo (flour mixed with molasses) once daily for the same period. Thyroid function was evaluated prior to initiation of treatment and after 8 weeks of treatment. Serum concentrations of total and free triiodothyronine (T3), total and free thyroxine (T4), and thyroid stimulating hormone (TSH) were determined at rest and after a thyrotropin-releasing hormone (TRH) stimulation test. There was no detectable difference between treatment and control groups.     

Evaluation of the thyroid status of Basenji dogs in Australia

Objective To determine the thyroid status of Basenji dogs in Australia. Methods Jugular or cephalic venipuncture blood samples were taken from 113 Basenji, comprising 47 males, 5 castrates, 48 entire and 13 spayed bitches, and sent on ice in plain and EDTA tubes to a single laboratory to determine haematocrit and serum concentrations of total thyroid hormone (thyroxine, TT4), thyroid‐stimulating hormone (TSH) and cholesterol. In a subgroup of 8 dogs with abnormal elevated TSH concentrations and subnormal TT4 concentrations, 5 were further examined by dynamic endocrine testing using recombinant human (rh) TSH (54 µg). Results Ages ranged from 1 to 14 years and weight range was 6.5 to 14.0 kg. TT4 concentrations (nmol/L) ranged from 2 to 27, with a median of 13 and a mean ± SD of 13.0 ± 5.7. Importantly, 85/113 (75%) of TT4 values were lower than the normal laboratory reference range (17–37). TSH concentrations (ng/mL) ranged from 0.05 to 5.37, with a median of 0.16 and a mean ± SD of 0.3 ± 0.6. Conclusions Basenji have a similar reference range for serum TSH, but a considerably lower reference range for TT4 (2–27 nmol/L) than most breeds and crossbreds, resembling the sight hounds in this respect. Given the difficulty of accurately measuring TT4 concentrations that are so low, concomitant serial TSH determinations are essential to properly asses thyroid function. Taken alone, TT4 determinations are only of use when the value is within the reference range, in which case a diagnosis of hypothyroidism is likely excluded.     

酮病奶牛血清中NO含量及NOS活性变化的研究

为了深入研究酮病的发生机理,选择荷斯坦奶牛作实验动物,应用酮粉法和改良式水杨醛比色法随机检测分组,Ⅰ组为10头阳性牛、Ⅱ组为10头对照牛。应用硝酸还原酶法测定血清中一氧化氮(NO)含量和一氧化氮合酶(NOS)活性。实验结果表明对照组奶牛血清中的NO含量及NOS活性均显著大于阳性组。由此可知,酮病可导致奶牛血清中NO含量降低、NOS活性下降。     

Effect of dietary Moringa oleifera leaves on the performance,ileal microbiota and antioxidative status of broiler chickens

In this study, we investigated the effect of Moringa oleifera leaves (MOL) supplements on the performance, ileal microbiota carcass traits and biochemical and antioxidant parameters of broiler chickens. MOL was added to starter, grower and finisher diets at four levels (0%, 0.5%, 1.0% and 5.0%). A total of 320 one-day-old mixed-sex Cobb-500 broiler chicks were divided equally into four treatment groups, with 5 replicates of 16 chicks each until 42 days of age. Compared to the control group, chicks fed 0.5% and 1% MOL exhibited a higher final body weight (FBW), greater weight gain (BWG), and better average daily gain (ADG). Feed intake (FI) gradually decreased as the inclusion rate of MOL in the broilers' diet increased to 5%. The best feed conversion ratio (FCR), protein efficiency ratio (PER) and performance index (PI) were observed in broilers that were fed the MOL diet. The percentage of abdominal fat in broilers decreased significantly in response to dietary supplementation with MOL. The haematological profile in the MOL group revealed a lower WBC count and lymphocyte percentage but higher heterophil and H/L ratio without affecting haemoglobin and RBC count. The inclusion of MOL increased total serum protein, Ca and P levels but decreased glucose, cholesterol, triglycerides, AST and ALT concentrations. Moreover, dietary MOL increased the activities of GSH, CAT and SOD, GPx and GST while decreasing the TBARS level. The pH value decreased in the ileum of broilers fed the 1% and 5% MOL diet compared to the other treatments. Decreased ileal counts of E. coli, Salmonella and Staphylococcus spp. were observed whereas the total ileal Lactobacillus spp. count increased in broilers fed the MOL diet. In conclusion, the MOL supplement can enhance growth performance and antioxidant status, alter ileal microbiota and protect against enteropathogens with no deleterious effects on the broiler chickens.     

γ-氨基丁酸对生长肥育猪抗氧化性能及HPA、HPT轴激素分泌的影响

研究基础日粮中添加10 mg/kg水平的γ-氨基丁酸(GABA)对生长肥育猪抗氧化性能及下丘脑-垂体-肾上腺皮质(HPA)、下丘脑-垂体-甲状腺(HPT)轴激素分泌的影响.选取48头体重43 kg左右的杜长大三元杂交猪随机分成2组,每组3个重复,每重复8头.分别饲喂基础日粮+0 mg/kg GABA(对照组)、基础日粮+10 mg/kgGABA(试验组).饲养试验结束时从每重复中随机选取2头(公母各半),共12头,前腔静脉采血制备血清后测定相关指标.结果表明:日粮添加10 mg/kg GABA使猪血清谷胱甘肽过氧化物酶(GSH-Px)活性极显著提高(P<0.01),同时超氧化物歧化酶(SOD)活性提高,且丙二醛(MDA)含量下降,但差异不显著(P0.05);显著降低猪HPA轴激素皮质醇、皮质酮及促肾上腺皮质激素(ACTH)水平(P<0.05);显著升高了(HPT)轴激素三碘甲腺原氨酸(T3)及游离三碘甲腺原氨酸(FT3)水平(P<0.05),但对甲状腺素(T4)及游离甲状腺素(FT4)无显著影响(P0.05).     

Effects of tiletamine-zolazepam anaesthesia on plasma antioxidative status and some haematological parameters in sheep

It is not clear whether the anaesthetic agents tiletamine and zolazepam have antioxidant or pro-oxidant effects. Therefore, this study was carried out to investigate the effects of tiletamine-zolazepam anaesthesia on oxidant/antioxidant status in blood plasma and on haematological parameters in 10 healthy Awassi ewes. The tiletamine-zolazepam combination was administrated in a dose of 7.5 mg/kg intramuscularly. The animals were spontaneously breathing air during the procedure. Blood samples were collected by jugular venipuncture before induction and at 30, 60, 120 min, 24 h and 3 days after anaesthesia. Malondialdehyde concentration, an index of lipid peroxidation, was higher at 30, 60, 120 min and 24 h (P < 0.05) than the baseline value in the plasma. The level of glutathione decreased (P < 0.05) at 30, 60 and 120 min, then returned to the baseline level. Beta-carotene concentration was lower (P < 0.05) than the baseline value during anaesthesia with the exception of its level at 120 min. Glutathione peroxidase and catalase activities decreased (P < 0.05) at the onset of anaesthesia, then returned to baseline values. There was no significant change in vitamin A level. Red blood cell count, haematocrit and haemoglobin concentration significantly decreased (P < 0.05) only at 30 min and thereafter they gradually returned to the baseline values. Based on the results tiletamine-zolazepam anaesthesia seems to accelerate lipid peroxidation and to impair the enzymatic antioxidant defence in the blood plasma.     

桑椹素对老龄小鼠肾阳虚症状的疗效试验

为进一步开发桑椹的药用保健功能,研究了桑椹素对老龄肾阳虚小鼠肾损伤的保护作用,并对其机制进行了探讨。将雄性老龄小鼠随机分为正常组和模型组,桑椹素低、中、高剂量组(0.1、0.2、0.4 g/kg)和补肾填精丸组(0.35 g/kg),测定小鼠血清、肾脏一氧化氮(NO)含量变化,观察小鼠肾脏组织结构病理变化。结果表明,模型小鼠以0.2、0.4 g/kg剂量桑椹素灌胃可明显改善肾阳虚症状,显著提高血清、肾脏NO含量,减轻氢化可的松引起的肾脏损害。其作用机制之一可能是通过提高肾脏NO的合成,增加肾血流量和肾小球滤过率,从而减轻对肾脏的损害。