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1.
NOS-immunoactive nerve structures were studied in the domestic hen (Gallus domesticus) stomach using single-labelling immunofluorescence. The examined tissues were collected from 10-12 months old laying hens (n=11), Isabrown and Zielononózka races. The distribution and number of the nerve fibres was examined considering differences between the proventriculus and gizzard, and also between particular layers of the stomach wall. Large or moderate numbers of nerve fibres showing NOS-immunoreactivity (NOS-IR) were found in all major layers of both parts of the stomach. In the muscular layer of the gizzard, NOS-positive nerve cell bodies were also observed. Small or moderate numbers of nerve terminals were distributed around blood vessels. NOS-IR nerve fibres associated with blood vessels were slightly more numerous in the gizzard than in the proventriculus. The widespread occurrence of NOS in both parts of the hen stomach can prove its important role in the control of digestive processes. It seems that there is a reasonable necessity of further, more detailed studies on morphological characteristics of the nitrergic nerve structures the bird alimentary tract, their functional significance and plasticity in different physiological and pathological conditions.  相似文献   

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This study examined the presence of immunoreactivity and mRNA for different nitric oxide synthase (NOS) isoforms in immature and in vitro matured oocytes and in embryos at two‐, four‐ and eight‐cell, and morula and blastocyst stages in buffalo. Oocytes obtained from slaughterhouse buffalo ovaries were subjected to in vitro maturation in TCM‐199 + 10% FBS + 5 μg/ml pFSH + 1 μg/ml estradiol‐17β + 0.81 mm sodium pyruvate + 10% buffalo follicular fluid + 50 μg/ml gentamycin sulphate for 24 h in a CO2 incubator (5% CO2 in air) at 38.5°C. Following in vitro fertilization carried out by incubating them with 2–4 million spermatozoa/ml for 18 h, the presumed zygotes were cultured in mCR2aa medium containing 0.6% BSA and 10% FBS for up to 8 days post insemination. Immunofluorescence staining of NOS using antibodies that cross‐reacted either with all the NOS isoforms i.e., universal (uNOS) or specifically with inducible (iNOS) or endothelial (eNOS) isoforms revealed that NOS was present in oocytes and embryos at all the stages examined. Examination of the semi‐quantitative expression of NOS genes by RT‐PCR revealed that the iNOS, eNOS and nNOS mRNA was present in the immature and mature oocytes and in all the embryonic stages examined. In conclusion, it was demonstrated in the present study that immunoreactivity and mRNA for different NOS isoforms was present in buffalo oocytes and pre‐implantation stage embryos.  相似文献   

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The present study was planned to evaluate a role of nitric oxide (NO) in the regulation of regular ruminal contractions in conscious sheep. Intravenous infusion of S-nitroso-acetyl-DL-penicillamine (SNAP) at doses of 3-30 nmol kg(-1) min(-1)for 30 minutes inhibited both the amplitude and frequency of ruminal contractions in a dose-dependent manner. However, intravenous infusion of Nomega-nitro-L-arginine-methyl ester (L-NAME) at doses of 0.3-3.0 micromol kg(-1) min(-1)did not alter the basal tone of intraruminal pressure and the amplitude of ruminal contractions. The frequency of contractions was slightly inhibited by L-NAME infusion at 1.0 micromol kg(-1)min(-1). The effects of L-NAME were abolished by simultaneous infusion of L -arginine at 30 micromol kg(-1) min(-1). These results suggest that exogenous NO can diminish the ruminal contractions, while endogenous NO is not involved in the regulatory mechanism of basal tone and regular phasic contractions of the rumen in healthy sheep.  相似文献   

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The aims of this study were to purify and localize the nitric oxide synthases (NOSs) from hybrid tilapia (Nile tilapia Oreochromis niloticus x Mozambique tilapia O. mossambicus). The purification procedures involved affinity chromatography with a 2', 5'-ADP-agarose 4B column and ion exchange with a diethylaminoethanol Bio-Gel A column. The results from gel filtration assays showed that the molecular weights of neuronal NOS (nNOS) and inducible NOS (iNOS) were 178 and 120 kDa, respectively. The sodium dodecyl sulfate-polyacrylamide gel electrophoresis results showed that there were three bands with molecular weights of 89, 47, and 29 kDa from the purified nNOS. However, only one band, with a molecular weight of 120 kDa, appeared on the gel from the purified iNOS. Hybrid tilapia nNOS was a dimer structure, while iNOS appeared to be a monomer structure. Moreover, our results revealed that the activities of nNOS and iNOS were significantly higher after the addition of Ca+2 or Mg+2 ions individually. However, when L-arginine and NADPH were present, the addition of 1 mM of either ion did not further increase the activity. The chemical L-N(G)-methyl-L-arginine could inhibit the activities of the purified NOSs with or without L-arginine. Western blot analyses showed only an 89-kDa immunoreactive band from the extracts of cerebrum; however, we did not find the specific bands in other tissues, such as gill, intestine, liver, spleen, and anterior kidney. We found another 120-kDa immunoreactive protein band with the rabbit antirat iNOS serum against iNOS from the extracts of anterior kidney and spleen. The results of immunohistochemistry with the rabbit antihuman nNOS serum indicated that the nNOS existed in the cerebellum, olfactory bulb, diencephalons, and nerve cell bodies and neuronal fibers of the spinal cord. Interestingly, only macrophages from anterior kidney and spleen showed positive reactions with the rabbit antirat iNOS serum. In the same way, the endothelial NOS (eNOS) located in the heart and epithelial cells of the blood vessels reacted positively with the rabbit antibovine eNOS serum.  相似文献   

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The objectives of this study were to evaluate the role of nitric oxide (NO) synthase isoforms (nNOS, eNOS, and iNOS) in the regulation of the migrating motor complex (MMC) in sheep using electromyography and their expression in the gastrointestinal (GI) tract by Western blot (WB) and immunohistochemistry. Intravenous administration of L-NAME or the nNOS inhibitor 7-nitroindazole (7-NI) decreased the MMC interval. Myoelectric activity of intestinal phase II was increased, whereas antral activity was reduced. These effects were blocked by L-arginine. Inhibitors of either iNOS (aminoguanidine and S-methylisothiourea) or eNOS (L-NIO) were ineffective. The NO donor sodium nitroprusside decreased GI myoelectric activity, inhibited the MMC pattern, and prevented the effects induced by L-NAME and 7-NI in the intestine. Intracerebroventricular administration of these agents did not modify GI motility. In the rumen, abomasal antrum, duodenum, and jejunum, WB showed three bands at about 155, 145, and 135kDa corresponding to nNOS, and a 140-kDa band (eNOS); however iNOS was not detected. Positive nNOS immunostaining was observed in neurons of the myenteric and submucous plexus of all GI tissues, while eNOS was found in the endothelial cells, ruminal and intestinal epithelium, as well as in some enteric neurons and in endocrine-like cells of the duodenal Brunner's glands. In contrast, only weak iNOS immunoreactivity was found in ruminal epithelium. Taken together, our results suggest that NO, synthesized at a peripheral level by nNOS, is tonically inhibiting the MMC pattern and intestinal motility in sheep.  相似文献   

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为探讨Balb/c小鼠正常生理状况下肺组织中一氧化氮自由基含量以及一氧化氮合酶活性的动态变化,采用电子自旋共振法直接测定了一氧化氮自由基含量,采用分光光度计法测定了一氧化氮合酶的活性.结果表明:在第3,6,7,12天Balb/c小鼠肺组织的一氧化氮自由基含量、一氧化氮合酶活性均无显著性差异(P>0.05).说明生理状态下,Balb/c小鼠肺组织一氧化氮自由基的产生维持动态平衡.  相似文献   

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一氧化氮 (NO)是一种最新发现的、哺乳动物中最小、最轻并具有独特理化性质和生物学活性的信息和效应分子 ,能激活靶细胞中的鸟苷酸环化酶 (GC) ,提高环一磷酸鸟苷(cGMP)浓度 ,发挥一系列生物学作用 ,现已成为研究热点之一。NO广泛存在于神经系统、心血管系统、免疫系统、消化系统、生殖系统与呼吸系统等的细胞内 ,是传递神经信息、调节血压以及机构防御等一系列生命活动必不可少的生物信使。因此 ,内源性NO的生物学研究、NO在动物模型大鼠体内的分布及其功能的确定 ,将有助于在动物医学和人类临床医学领域进一步阐明机体某些…  相似文献   

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为了研究D-半乳糖联合铝诱导的小鼠阿尔茨海默症(AD)脑海马一氧化氮合酶(NOS)活性和一氧化氮(NO)含量的变化及L-NNA和盐酸多奈哌齐对其变化的影响,探讨NO在阿尔茨海默症中的作用机制及2种药物对脑神经元的保护作用。选取2月龄健康昆明小鼠160只,体质量(20±2)g,随机分为正常对照组、模型组、盐酸多奈哌齐治疗组及L-NNA治疗组,利用D-半乳糖联合三氯化铝建立小鼠AD模型,应用生化检测技术测定各组脑海马在造模后每周NOS活性及NO含量。结果表明,模型组海马内NOS活性开始呈缓慢升高,从第4周开始呈显著升高,保持较高含量至12w造模结束;两治疗组脑海马NOS活性在各时间点极显著或显著低于模型组(P0.01或P0.05),并且L-NNA在4~8周时降低脑海马NOS活性的程度好于盐酸多奈哌齐治疗组(P0.05);NO含量的变化随着NOS活性的变化而变化;利用SABC免疫组织化学方法检测各组脑海马神经型NOS(nNOS)阳性神经元,发现模型组脑海马nNOS阳性神经元密度降低,细胞着色变淡,胞体截面积和最长突起长度变小,经过治疗后神经元密度增加,胞体截面积和最长突起长度显著改善(P0.05)。结果提示NO参与了AD形成过程,高浓度的NO能发挥神经毒性作用损害脑组织;L-NNA通过抑制NOS的活性,降低了脑海马NO的含量,对阿尔茨海默症中海马神经元具有明显的保护作用。  相似文献   

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一氧化氮 (NO)作为内皮源性舒张因子(EDRF)的主要形式 ,在心血管系统的生理和病理生理中发挥着重要作用。研究表明 :肺动脉高压和心衰的发生与 NO的代谢紊乱有关 ,有文献报道 NO可产生负性肌力作用、增强舒张功能 ;也有人认为NO对心功能无影响 ,但 NO能降低肺动脉高压和高血压的发生。内皮素是最强烈的血管收缩因子 ,两者由内皮细胞合成 ,血管内皮细胞功能失调与肺心血管疾病的发生、发展有关 ,其中最重要的疾病是肺动脉高压和原发性高血压 [1,2 ]。病理状态下 ,由于血流切应力及血流搏动过强刺激血管内皮细胞 ,造成其功能失调 :血管…  相似文献   

11.
The objective of the present study was to investigate the influence of prostaglandin F(2alpha) (PGF (2alpha)) and nitric oxide (NO) on production of steroids and PGs by culturing bovine luteal cells obtained from ovaries on days 8-12 of the estrous cycle with a nitric oxide (NO) donor (Spermine NONOate), and a NO synthase inhibitor (N(G)-nitro-L-arginine methyl ester dihydrochloride: L-NAME). When the cells were exposed for 24 h to PGF(2alpha) (10(-7)-10(-5) M), production of progesterone (P(4)) increased significantly at all doses used (P<0.05). Moreover, PGF(2alpha) stimulated PGF(2alpha) production (P<0.01), depressed testosterone (T) production (P<0.05), but did not affect synthesis of prostaglandin E(2) (PGE(2)). Spermine NONOate decreased P(4) production to 66%, 47% and 34% of the control concentration after treatment with 10(-5) M, 10(-4) M and 10(-3) M, respectively, but did not affect T production, and increased PGF(2alpha) synthesis (P<0.05) and PGE(2) (P<0.01) at all doses used. L-NAME increased production of P(4) (P<0.01) but did not affect (P>0.05) secretion of T, PGF(2alpha) and PGE(2). Estradiol-17beta (E(2)) was detectable on the level of sensitivity of assay and was not significantly altered by any treatments. The overall results suggest that PGF(2alpha) and NO produced locally in bovine CL play roles in the regulation of the secretory function of the bovine CL as auto/paracrine factors.  相似文献   

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铜是动物体内一种必需的微量元素,在多种代谢途径中有重要的生物学作用.早期的研究结果表明,提高饲料中铜的含量,可以促进动物生长.但是过量添加铜也可以损害肝脏、肾脏等器官[1-2].一氧化氮(NO)作为一种强有力的内皮细胞舒血管因子,其功能已越来越受到人们的重视.NO对肾脏具有保护和损伤的双重作用.一方面具有扩张动脉,抑制血小板粘附、聚集,抗血栓形成,疏通微循环,增加供血,保护细胞的作用.  相似文献   

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Inflammation is a process consisting of a complex of cytological and chemical reactions which occur in and around affected blood vessels and adjacent tissues in response to an injury caused by a physical, chemical or biological insult. Much work has been performed in the past several years investigating inducible nitric oxide synthase (NOS, EC 1.14.13.39) and nitric oxide in inflammation. This has resulted in a rapid increase in knowledge about iNOS and nitric oxide. Nitric oxide formation from inducible NOS is regulated by numerous inflammatory mediators, often with contradictory effects, depending upon the type and duration of the inflammatory insult. Equine medicine appears to have benefited the most from the increased interest in this small, inflammatory mediator. Most of the information on nitric oxide in traditional veterinary species has been produced using models or naturally occurring inflammatory diseases of this species.  相似文献   

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A vast majority of the world buffalo resource is concentrated in tropical and subtropical countries. Apart from heat stress and poor nutritional availability, endometritis is one of the most commonly encountered reproductive problems limiting fertility and consequently productive potential of the species. As demonstrated recently, endometritis impairs growth and follicular fluid composition of the largest follicle in buffalo. In the present study, the effect of endometritis on luteal development, function, nitric oxide (NO), and ascorbic acid was investigated. Reproductive tracts were collected from 90 cyclic buffaloes at an abattoir and grouped into endometritic (n?=?36) or non-endometritic (n?=?54) buffaloes based on physical examination of uterine mucus, white side test, and uterine cytology. Samples with pus-containing mucus, positive reaction on white side test, and/or >5 % neutrophils were considered to be positive for endometritis. Corpora lutea were enucleated, weighed, classified into stages I to IV, and assayed for progesterone (P4), NO, and ascorbic acid concentrations. Endometritic buffaloes had lesser (P?<?0.0001) luteal weight and P4, NO, and ascorbic acid concentrations than non-endometritic buffaloes. The findings indicated that endometritis impairs corpus luteum development and function in buffalo. Reduced luteal NO and ascorbic acid concentrations during endometritis are novel findings.  相似文献   

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The objective of this study was to examine the follicular characteristics and intrafollicular concentrations of nitric oxide and ascorbic acid during ovarian acyclicity in buffaloes. Ovaries were collected from 56 acyclic and 95 cyclic buffaloes at slaughter, surface follicle number was counted and follicles were classified into small (5.0–6.9 mm), medium (7.0–9.9 mm), and large (≥10.0 mm) size categories based on their diameter. Follicular fluid was aspirated and assayed for nitric oxide, ascorbic acid, estradiol, and progesterone. Acyclic buffaloes had a higher (P < 0.05) number of medium-sized follicles and a lower (P < 0.001) number of large follicles than the cyclic ones. In acyclic animals, the number of large follicles was lower (P < 0.01) than in medium size category which in turn was lower (P < 0.001) than the number of small follicles. In contrast, the number of medium and large follicles was not different (P > 0.05) in the cyclic control. However, the number of small-sized follicles was higher (P < 0.001) compared to the other two categories. The incidence of large-sized follicles was lower (P < 0.05) in acyclic buffalo population compared to the cyclic control. Evaluation of estrogenic status demonstrated that all the follicles of acyclic buffaloes are estrogen-inactive (E 2/P 4 ratio < 1). Small- and medium-sized follicles of acyclic buffaloes had higher concentrations of nitric oxide (P < 0.05 and P < 0.001, respectively) and lower concentrations of ascorbic acid (P < 0.05 and P < 0.01, respectively) than the corresponding size estrogen-active follicles of their cyclic counterparts. In conclusion, this study indicates that follicular development continues during acyclicity in buffaloes. Although follicles in some acyclic buffaloes attain a size corresponding to morphological dominance, they are unable to achieve functional dominance, perhaps due to an altered balance of intrafollicular nitric oxide and ascorbic acid and, as a result, these follicles instead of progressing to ovulation undergo atresia.  相似文献   

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In this study we investigated the effect of kefir on the levels of glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO) in the liver, stomach, spleen and colon of mice with colonic aberrant crypts formed by azoxymethane (AOM). Thirty 12 weeks old Swiss Albino mice averaging 31.5 g weight were used as experimental animals. The mice were separated into 3 groups. The first group was the control group, second group was the AOM and third group was the AOM+kefir group. We applied AOM to the second and third groups. Mice were fed ad libitum by laboratory rodent chow during the experiment period. Water was given to the first and second groups and third group received only kefir diluted with water (50%). AOM was injected subcutaneously to the second and third groups for 7 weeks (two times a week, 5 mg/kg). Six weeks after the final AOM treatment the animals were sacrificed and liver, stomach, spleen and colon samples were collected from all the groups. MDA level demonstrated an increase only in stomach for the third group (p < 0.001), while an elevation was observed for all of the four organs for the second group (spleen p < 0.001, liver p < 0.001, colon p < 0.01). GSH level showed an increase in the second group at stomach (p < 0.01) and colon (p < 0.001), while in the third group, a small increase was determined only at the colon (p < 0.05). NO level increased at all of the organs in the second group (spleen, liver, colon p < 0.001, stomach p < 0.05), but only at liver and colon in the third group 3 (p < 0.001). In conclusion these results showed that kefir plays an antioxidant role.  相似文献   

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