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.     

Role of G-protein-coupled receptor kinases in cell migration and signal transduction

G-protein-coupled receptor kinases (GRKs) are a family of serine/threonine protein kinases. The investigators pay much attention to the roles of GRKs in the signal transduction through G-protein-coupled receptors (GPCRs) with arrestin ever since a long time ago. Due to the physiological and pathological observations with the methods of deletion or overexpression, GRKs are considered as new drug targets. The kinases play a role in the pathogenesis of hypertension and cell migration through GPCRs and Hedgehog signaling pathways. As the development of research techniques, especially bioluminescence resonance energy transfer (BRET) and fluorescence resonance energy transfer (FRET), the special mechanism of GRKs for GPCRs is more evident. In this review, we discuss the recent achievement in the roles of GRKs signaling and the related newest research techniques.     

Binding of canine anaphylactic antibodies on human basophils: application to canine allergy diagnosis

Abstract Using the technique of human basophil passive sensitization, as employed for human allergy díagnosis, we checked the ability of canine anaphylactic antibodies to sensitize human basophils. Therefore, by sensitizing human basophils with sera taken from dogs allergic to house dust mite, we demonstrated basophil activation as measured by alcian blue staining. Basophil activation was inhibited by heating dog sera at 56 °C for 6 h and by a human myeloma IgE. Basophil activation was also shown by histamine and leukotriene (LTC4) release. These results indicate canine anaphylactic antibodies bind to human basophil IgE receptors and also that they are IgE. The three methods described here for measuring basophil activation may lead to díagnostic methods applicable to canine allergy díagnosis. Resumen Mediante el método de la sensibilización pasiva de basófilos humanos como se utiliza para el díagnóstico de la alergia humana, evaluamos la capacidad de los anticuerpos anafilácticos caninos de sensibilizar basófilos humanos. Asi, sensibilizando basófilos humanos con suero extraido de perros con alergia al ácaro del polvo, demostramos la activación de basófilos medíante la tinción de Azul de Alcián. Se inhibió calentando suero canino a 56 °C durante 6 h. y por IgE de mieloma humano. La activación de los basófilos se mostró también por la liberación de histamina y leucotrieno (LTC4). Estos resultados indican que los anticuerpos caninos anafilácticos se unen a los receptores de IgE en basófilos humanos y también que son IgE. Los tres métodos descritos aqui para medir la activación de basófilos pueden llevar a métodos de díagnóstico aplicables al díagnóstico de la alergia canina. [Sainte-Laudy, J., Prost, C. Binding of canine anaphylactic antibodies on human basophils: application to canine allergy díagnosis (Union de anticuerpos anafilácticos caninos a basófilos humanos: aplicacion al díagnóstico de alergia canina). Veterinary Dermatology 1996; 7 : 185–91.] Résumé Utilisant une technique de sensibilisation passive de basophiles humains, employee pour le díagnostic allergologique chez l'homme, nous avons testé la capacité des anticorps anaphylactiques canins à sensibiliser des basophiles humains. Ainsi, par sensibilisation de basophiles humains avec des sérums provenant de chiens allergiques aux acariens de la poussière de maison, nous avons démontré l'activation des basophiles mesurée par coloration au bleu alcian. Celle-ci est inhibée par des sérums canins chauffés à 56 °C pendant 6 heures et par un myélome IgE humain. L'activation des basophiles a été aussi démontrée par libération d'histamine et de leucotriénes (LTC4). Ces résultats prouvent la présence d'anticorps anaphylactiques canins fixés à des récepteurs IgE de basophiles humains et que ceux-ci sont des IgE. Les trois méthodes décrites ici pour mesurer l'activation des basophiles peuvent être utilisées pour le díagnostic allergologique chez le chien. [Sainte-Laudy, J., Prost, C. Binding of canine anaphylactic antibodies on human basophils: application to canine allergy díagnosis (Fixation d'anticorps anaphylactiques canins sur des basophiles humains). Veterinary Dermatology 1996; 7 : 185–91.] Zusammenfassung Mit der Technik der passiven Basophilensensibilisierung beim Menschen, wie man sie für die Allergiedíagnose beim Menschen anwendet, untersuchten wir die Möglichkeit, menschliche Basophile durch kanine anaphylaktische Antikörper zu sensibilisieren. Dazu wurden humane Basophile mit Sera von Hunden sensibilisiert, die allergisch auf Hausstaubmilben reagierten. Dabei demonstrierten wir eine Basophilenaktivierung, die durch Elsässerblau-Färbung gemessen werden konnte. Der Vorgang wurde verhindert durch Erhitzen der Hundesera auf 56 °C für 6 Stunden und durch humanes Myelom-IgE. Basophilenaktivierung wurde auch durch Histamin-und Leukotrien(LTC4)-Ausschüttung gezeigt. Diese Ergebnisse zeigen, daß kanine anaphylaktische Antikörper sich an humane basophile IgE-Rezeptoren binden und auch IgEs darstellen. Die drei hier beschriebenen Methoden zur Messung der Basophilenaktivierung können zu einer díagnostischen Methode führen, die für die Diagnostik kaniner Allergie anwendbar ist. [Sainte-Laudy, J., Prost, C. Binding of canine anaphylactic antibodies on human basophils: application to canine allergy díagnosis (Die Bindung von anaphylaktischen Antikörpern des Hundes an Basophile Zellen des Menschen: Anwendung für die Allergiedíagnose beim Hund). Veterinary Dermatology 1996; 7 : 185–91.]     

Immunolocalization of sex steroid receptors in the epididymis and ductus deferens of immature and mature Japanese Quail, Coturnix Japonica

The goal of this study was to determine whether in the Japanese quail the male genital tract contains receptors for progesterone, androgen and estrogen (PR, AR and ER, respectively), which have significant roles in reproductive functions, and whether their localization changes during sexual maturation. The epididymis and ductus deferens (middle and ampulla regions) of immature (approximately 30-day-old) and mature male Japanese quail were collected and frozen sections of them were immunostained for PR, AR and ER. The immunoreaction products for AR and PR were found in the nuclei of epithelial cells in the efferent ductules, epididymal duct, and the middle and ampulla regions of the ductus deferens of mature and immature birds. In the mature birds, the epithelial cells of the efferent ductules, epididymal duct, and the middle and ampulla regions of the ductus deferens were positive for ER, although some of the cells in the ductus deferens were negative. The epithelial cells of the ductules in the epididymis stained positive for ER, but the immunoreactions were negligible in the ductus deferens of immature birds. These results suggest that the epididymis and ductus deferens in quail possesses PR, AR and ER receptors. Each receptor is expressed before sexual maturation, although enhancement of ER expression may occur during maturation.     

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.     

Roles of atypical chemokine receptors in progression and metastasis of tumor

Chemokines and their receptors have been implicated mostly in tumor progression and metastasis. Atypical chemokine receptors (ACKRs) comprise a group of 7-transmembrane domain proteins structurally similar to G protein-coupled receptors. However, ACKRs do not induce classical signaling via the typical G protein-mediated pathways. ACKRs efficiently internalize the cognate chemokine ligands and act as scavengers instead. ACJRs are composed of at least 3 members of chemokine receptors:Duffy antigen receptor for chemokines (DARC, also known as ACKR1), D6 (also known as ACKR2) and ChemoCentryx chemokine receptor (CCX-CKR, also known as ACKR4). These receptors bind to and/or internalize their chemoattractant ligands without activating signal transduction cascades leading to cell migration. In this review, we summarize the recent progress regarding the roles of ACKRs in the progression and metastasis of tumor.     

Protective effect of adiponectin on glucose and lipid metabolism by suppressing MHCⅡ expression

AIM: To investigate whether the protective effect of adiponectin on glucose and lipid metabolism is achieved through down-regulating major histocompatibility complex class Ⅱ (MHCⅡ) in the adipose tissue. METHODS: Adiponectin knockout (KO) mice and C57BL/6(WT) mice were fed with high-fat diet and standard diet for 24 weeks, respectively. The body weight, fasting blood glucose (FBG), fasting insulin (FINS), homeostasis model assessment of insulin resistance (HOMA-IR), triglyceride (TG), total cholesterol (TC), low-density lipoprotein cholesterol (LDL-C), high-density lipoprotein cholesterol (HDL-C), hepatic histology, and class Ⅱ trans-activator (CⅡTA), histocompatibility 2 class Ⅱ antigen E beta (H2-Eb1) and cluster of differentiation 74(CD74) mRNA and MHC Ⅱ protein levels in adipose tissue were measured at sacrifice. siRNA targeting MHC Ⅱ and overexpression vector was used in 3T3-L1 cells to explore the effect of adiponectin on the protein level of MHCⅡ. RESULTS: The levels of body weight, FBG, FINS, HOMA-IR, TC, TG, LDL-C, hepatic steatosis, CⅡTA, H2-Eb1 and CD74 mRNA expression, and MHCⅡ protein expression in the KO mice were higher than those in the WT mice that fed with high-fat diet or standard diet. In 3T3-L1 cells, inhibition of adiponectin reversed MHC Ⅱ protein level induced by specific siRNA. The expression of MHC Ⅱ in adipocytes decreased after adiponectin was overexpressed. CONCLUSION: Adiponectin improves glucose and lipid metabolism through suppressing the expression of MHCⅡ in the adipose tissue.     

Pharmacological and therapeutic targets for Δ9 tetrahydrocannabinol and cannabidiol

Summary Cannabis is the unique source of a set of at least 66 compounds known collectively as cannabinoids. Of these, most is known about the pharmacology of ⁹-tetrahydrocannabinol (⁹-THC), the main psychoactive constituent of cannabis, and about cannabidiol (CBD), which lacks psychoactivity. Accordingly, this paper focuses on the pharmacological and therapeutic targets of these two cannabinoids. Many of the effects of ⁹-THC are mediated by cannabinoid receptors of which at least two types, CB₁ and CB₂, are present in mammalian tissues. Endogenous agonists for cannabinoid receptors have also been discovered. CB₁ receptors are present at the terminals of central and peripheral neurones, where they modulate transmitter release. They also exist in some non-neuronal cells. CB₂ receptors are expressed mainly by immune cells, one of their roles being to alter cytokine release. ⁹-THC also appears to have non-CB₁, non-CB₂ pharmacological targets. It is already licensed for clinical use in the U.S.A. as an anti-emetic and appetite stimulant and both ⁹-THC and ⁹-THC-rich cannabis extracts show therapeutic potential as neuroprotective and anticancer agents and for the management of glaucoma, pain and various kinds of motor dysfunction associated, for example, with multiple sclerosis and spinal cord injury. CBD has much less affinity for CB₁ and CB₂ receptors than ⁹-THC and its pharmacological actions have been less well characterized. Potential clinical applications of CBD and CBD-rich cannabis extracts include the production of anti-inflammatory and neuroprotective effects, the management of epilepsy, anxiety disorders, glaucoma and nausea, and the modulation of some effects of ⁹-THC.     

口蹄疫流行病学及感染机制的研究进展

口蹄疫(Foot-and-mouth disease FMD)是由口蹄疫病毒(Foot-and-mouth disease virus FMDV)引起偶蹄动物的一种急性、热性、高度接触性传染病。FMDV有多种血清型及其亚型,目前主要引起动物致病的就有7种,即A、O、C、SATⅠ、SATⅡ、SATⅢ和亚洲Ⅰ型(AsiaⅠ型)。病毒通常在细胞受体的参与下才入侵宿主细胞,进行扩增生命循环,感染宿主细胞,使得该病得以广泛流行。但由于口蹄疫病毒的高度变异性和适应性而在流行中出现新的特点,以及被发现的病毒在动物体内和细胞中长期存在而引发FMDV的持续感染等问题,使得本病不能有效被控制而在许多国家和地区重新暴发和流行。为此本文将口蹄疫病原学、流行病学、病毒细胞受体及其病毒在体内持续感染形成的原因方面进行的论述,为口蹄疫病毒感染机制的研究提供了科学的依据。     

成年大鼠睾丸去精索上神经对睾丸功能及肾上腺素能受体表达的影响

试验旨在研究精索上神经（superior spermatic nerve,SSN）对睾丸功能的影响,25只成年Wistar大鼠随机分为2组,试验组左右睾丸切除精索上神经,手术30 d后两组大鼠同时处死取样分析。结果显示,睾丸去精索上神经不影响睾丸指数的大小但使附睾尾精子数极显著降低（P＜0.01）,平均日增重显著增加（P＜0.05）。增殖细胞核抗原（PCNA）免疫组织化学染色结果显示,睾丸去精索上神经不影响曲细精管精原细胞和初级精母细胞的增殖,但影响曲细精管的形态和生殖细胞的规则排列。RT-PCR结果显示,睾丸去精索上神经明显上调β1AR mRNA的表达,下调β2AR mRNA的表达;另外切除精索上神经不影响睾丸3β-羟胆固醇脱氢酶（3β-HSD）mRNA表达,但显著抑制类固醇快速调节蛋白（StAR）、胆固醇侧链裂解酶（P450scc）mRNA表达（P＜0.05）。结果表明,支配睾丸的精索上神经通过调节肾上腺素能受体β1AR和β2AR的表达及StAR和P450scc影响睾丸睾酮的合成和精子的发生。