Inhibition of histamine release from dispersed canine skin mast cells by cyclosporin A, rolipram and salbutamol, but not by dexamethasone or sodium cromoglycate

Despite the important role that canine skin mast cells play in IgE-mediated allergic inflammation, clinically useful compounds for modulating mediator release from these cells or for suppressing cell response are lacking in the dog. The ability of five compounds to inhibit histamine release induced by non immunological (calcium ionophore A23187 and substance P) and IgE-dependent (concanavalin A) stimuli were compared. Sodium cromoglycate, a mast cell stabilizer, and dexamethasone, a glucocorticoid, failed to inhibit histamine release from isolated skin mast cells following any kind of stimulation. Salbutamol, a β-adrenergic agonist, exhibited inhibitory activity (46.0%) only after concanavalin A activation. In contrast, rolipram, a selective phosphodiesterase IV inhibitor and cyclosporin A, an immunosuppressor, showed potent anti allergic actions, inhibiting both IgE-dependent and -independent stimuli. Rolipram inhibited 42.8%, 44.7% and 19.2% of the mediator release induced by ionophore A23187, substance P and concanavalin A, respectively. Similarly cyclosporin A induced 85.9%, 14.9% and 67.3% inhibition after ionophore A23187, substance P and concanavalin A stimulation, respectively. These results suggest that rolipram and cyclosporin A merit to be clinically tested as agents for the treatment of chronic allergic diseases in the dog.     

Role of PPARα/PGC-1α in doxorubicin induced mouse dilated cardio-myopathy

AIM: To investigate the changes of peroxisome proliferator-activated receptors (PPAR)α/peroxisome proliferator activated receptor coactivator 1 alpha (PGC-1α) in doxorubicin (DOX) induced dilated cardiomyopathy (DCM) and its effect on the energy metabolism and myocardial function in mice. METHODS: Forty mice were randomly divided into 4 groups: control group, DOX group, PPARα inhibitor group and PPARα agonist group. The DCM model was established by injection of DOX. The protein levels of PPARα/PGC-1α were detected. The PPARα inhibitor and PPARα agonist were used 2 weeks beforeinjection of DOX. The contents of adenine acid and phosphocreatine (Pcr) in the mitochondria were measured by high-performance liquid chromatography (HPLC). The ANT activity was analyzed by the atractyloside-inhibitor stop technique. The changes of the echocardiography and hemodynamics were also observed. RESULTS: DOX induced DCM model was successfully established. The protein levels of PPARα and PGC-1α in control group were significantly higher than those in DOX group (P<0.05). Both of the high-energy phosphate contents and the transport activity of ANT were decreased in DOX group (P<0.05), and the hemodynamic parameters were disordered (P<0.01). Compared with DOX group, PPARα inhibitor pre-treatment significantly reduced the PPARα/PGC-1α expression. Meanwhile, high-energy phosphate contents in the mitochondria and the ANT transport activity of the mitochondria decreased, as well as the left ventricular function (P<0.05). On the other hand, PPARα agonist significantly increased the expression of PPARα and PGC-1α, and improved the transport activity of ANT. In addition, the hemodynamic parameters were ameliorated, but the high-energy phosphate contents of the mitochondria did not significantly change. CONCLUSION: PPARα/PGC-1α plays an important role in the regulation of ANT transport activity in dilated cardiomyopathy induced by DOX, and the activation of PPARα/PGC-1α has protective effects on the DCM induced by DOX.     

水貂部分Toll样受体(TLRs)基因的分子克隆、序列分析及组织表达分析

试验旨在研究Toll 样受体(Toll-like receptors,TLRs)在水貂抗病毒免疫中的作用机制,并为抗病育种积累可供选择的基因素材。本试验以雪貂TLRs基因为参考序列设计4对引物对水貂TLR4、TLR6、TLR7及TLR8进行分子克隆,并对所得序列进行生物信息学分析,进一步利用半定量RT-PCR技术分析TLR4和TLR7基因mRNA在不同年龄水貂各组织中的表达情况。结果表明,水貂TLRs与食肉目动物(雪貂、北极熊、大熊猫、海象、海豹、犬、老虎和猫)具有较高的同源性,在系统发育树中距离最近;组织表达分析表明TLR4和TLR7在检测的组织中广泛表达且表达量存在差异,相比成年水貂,仔貂各组织中TLR4或TLR7基因表达量更高。     

Toll-like receptor-mediated anti-inflammatory action of glaucine and oxoglaucine

Two isochinoline alkaloids, glaucine and oxoglaucine were investigated for their suggested anti-inflammatory influence concerning nitric oxide and cytokine production. Mouse peritoneal macrophages were stimulated with different Toll-like receptor (TLR) ligands such as LPS for TLR4, zymosan for TLR2 and CpG for TLR9. The alkaloids inhibited TNF-α and IL-6 production induced by these ligands. In regard to IL-12 suppressive effect was registered in the case of CpG stimulation. Glaucine succeeded to enhance LPS and zymosan-induced IL-10 production. The reduction of pro-inflammatory cytokines and increase of anti-inflammatory IL-10 are indicative for their use in different acute and chronic inflammatory diseases.     

褪黑素受体MT1和MT2在绵羊胃中的分布及表达

【目的】应用分子生物学技术探究绵羊不同胃室组织褪黑素(MT)特异性膜受体MT1和MT2的分布规律及表达模式,以初步探明绵羊不同胃室组织褪黑素调节胃消化的生物效应机制。【方法】采集绵羊瘤胃背囊、瘤胃腹囊、网胃、瓣胃和皱胃5个组织部位,用ELISA、实时荧光定量PCR、免疫组化和Western blotting方法检测褪黑素特异性膜受体MT1和MT2在绵羊胃组织不同部位的分布规律及表达模式。【结果】ELISA结果显示,绵羊各胃组织中均含有褪黑素,且皱胃中褪黑素含量最高,瓣胃次之,瘤胃背囊、瘤胃腹囊和网胃中均较低。实时荧光定量PCR结果显示,MT1和MT2基因mRNA在皱胃中含量均最高,其次是瘤胃腹囊,在瘤胃背囊和网胃中含量较低。免疫组化结果显示,MT1和MT2蛋白在绵羊各胃组织中均有分布,主要表达于各胃组织的黏膜层,且在皱胃腺体中的分布呈从底部到颈部逐渐增多的趋势。Western blotting结果显示,MT1和MT2蛋白在网胃中表达均最高,瓣胃次之,在瘤胃背囊、瘤胃腹囊和皱胃中表达均较低。【结论】褪黑素在绵羊各胃组织中差异化表达从而发挥多样性生理功能,可能通过与胃组织上特异性受体MT1和MT2结合,通过信号传导系统而调控前胃受食糜刺激后发生反刍生理过程。     

Orexin and orexin receptor like peptides in the gastroenteric tract of Gallus domesticus: An immunohistochemical survey on presence and distribution

This study reports the immunohistochemical localization and distribution of orexin A and B-like and their receptors-like peptides in the gastroenteric tract of chicken. The immunoreactivity is distributed in endocrine cells, nerve fibers and neurons, both in the stomach and intestine, and shows a discrete conformity with the data till now reported for Mammals. Our study suggests a possible participation of orexin-like peptides in the modulation of chicken gastroenteric activities and the preservation of their main distribution compared to Mammals. Western blot analysis has confirmed the presence of prepro-orexin and both receptors in the examined tissues.     

Effects of modulatory agents on neurally-mediated responses of trout intestinal smooth musclein vitro

Mediators and mechanisms responsible for the inhibitory modulation of trout intestinal smooth muscle were examined using a series of putative mediators and substances known to modulate neurotransmission in mammalian systems. Frequency response relationships to transmural stimulation and concentration response relationships to 5-hydroxytryptamine, carbachol, and substance P were established on paired segments of rainbow trout intestinein vitro in the presence and absence of putative modulatory agents. Modulation of neurally-mediated contractions of trout intestine was achieved with dibutyryl cyclic AMP and forskolin, agents that increase intracellular levels of cyclic AMP. The effect appears to be at the level of the smooth muscle, since the adenylate cyclase activator, forskolin, inhibited muscarinic and serotoninergic contractions as well as transmurally stimulated contractions. Substance P-induced contractions were unaffected by forskolin. The endogenous agonists/neurotransmitters which would increase cyclic AMP levels in rainbow trout intestinal smooth muscle are as yet unknown. The effects do not appear to be modulated by vasoactive intestinal peptide (VIP), calcitonin, calcitonin gene-related peptide (CGRP), or agents that activate -adrenoceptors. Prostaglandin E₂ (PGE₂) and ₂-adrenergenic agonists are possible agents which will decrease contractility of the smooth muscle. They were only active in the proximal intestine and on transmurally stimulated contractions. The effects of both PGE₂ and ₂-agonists appear to be prejunctional, decreasing release of contractile neurotransmitters in the enteric nervous system.     

大豆苷元对断奶仔猪胃肠道发育的影响

试验选用3窝新生仔猪,每窝内随机分为两组:大豆苷元组和对照组,于7、9、11日龄每头仔猪分别饲喂1、2和3mg.mL-1大豆苷元脱脂乳溶液1mL或等体积脱脂乳;所有仔猪于21日龄断奶;于14、21、24和35日龄,每窝随机抽取大豆苷元组和对照组仔猪各一头称重,屠宰,称量脾脏和胸腺重;取胃底部、十二指肠前端、空肠前端用于组织形态学观察;测定空肠和回肠组织组胺以及血清皮质醇含量。结果表明:大豆苷元显著增加断奶仔猪胃底腺盐酸细胞数量(P<0.05),但对胃底腺厚度和肠绒毛高度无显著影响;大豆苷元既可以阻止断奶造成的胃底腺肥大细胞募集(P<0.05),又可以维持肥大细胞的平稳增加;此外大豆苷元还可以降低断奶后回肠组胺水平(P<0.05);皮质醇在所有采样时间都无显著差异,说明大豆苷元对肥大细胞和组胺的作用可能与皮质醇无关;除35日龄大豆苷元组胸腺重高于对照组(P<0.05)外,大豆苷元对断奶仔猪胸腺和脾脏质量无显著影响。