不同电针对抑郁模型大鼠结肠黏膜脑肠肽GAS、NPY、CGRP含量的影响

目的观察不同电针"百会""印堂"对抑郁模型大鼠结肠黏膜三种胃肠道相关脑肠肽胃泌素(GAS)、神经肽Y(NPY)及降钙素基因相关肽(CGRP)的影响,为揭示针刺治疗抑郁症胃肠道躯体化症状作用途径提供实验依据,并比较不同电针的作用效果。方法将60只健康SD大鼠随机分为空白组、模型组、百优解组、脉冲电针组、音乐电针组,每组12只。采用慢性不可预见的温和刺激结合孤养方法制备抑郁模型。百忧解组每天将百忧解以生理盐水配制灌胃给药。脉冲电针组针刺大鼠"百会"和"印堂"穴,接脉冲电针治疗仪。音乐电针组取穴同脉冲点针组,接音乐电针治疗仪。采用放射免疫法测定各组大鼠结肠黏膜GAS、NPY、CGRP含量。结果模型组与空白组相比,大鼠结肠黏膜GAS含量显著降低(P<0.05),NPY含量显著升高(P<0.05),CGRP含量极显著升高(P<0.01);脉冲电针组、音乐电针组及百忧解组与模型组相比,大鼠结肠黏膜GAS含量显著升高(P<0.05或P<0.01),NPY含量显著降低(P<0.05或P<0.01),CGRP含量显著降低(P<0.05或P<0.01);音乐电针组与脉冲电针组相比,大鼠结肠黏膜CGRP含量降低水平更为显著(P<0.05)。结论电针治疗可通过调整抑郁后胃肠道兴奋性激素与抑制性激素的失平衡来影响肠道中脑肠肽含量,从而调控脑-肠轴,使抑郁后胃肠功能紊乱状态得到改善,而且音乐电针组优于脉冲电针组。     

Opposing effects of delta- and zeta-protein kinase C isozymes on cardiac fibroblast proliferation: use of isozyme-selective inhibitors

Neonatal primary cardiac fibroblasts in defined medium continue to proliferate. Here, we show that phorbol ester inhibited and transforming growth factor-beta1 (TGFbeta1) stimulated this fibroblast proliferation. Cardiac fibroblasts contain six protein kinase C (PKC) isozymes: alpha-, delta-, epsilon-, betaI-, betaII-, and zeta-PKC. To evaluate the effect of different PKC isozymes on the proliferation of these cells, we used isozyme-selective PKC inhibitors. Inhibition of endogenous delta-PKC with deltaV1-1, an isozyme-selective translocation inhibitor, resulted in increased basal thymidine incorporation by 58 +/- 12% of control cells, but did not affect TGFbeta1-induced cell growth. Inhibition of endogenous zeta-PKC in neonatal rat cardiac fibroblasts with zeta-pseudosubstrate, a selective inhibitor for the atypical PKC isozymes, revealed an opposite effect; this inhibitor reduced basal growth to 45 +/- 11% and TGFbeta1-induced growth to 61 +/- 10%. Other isozyme-specific inhibitors used in this study did not alter basal or TGFbeta1-stimulated fibroblast growth. Taken together, our data provide evidence that delta-PKC inhibits and zeta-PKC stimulates proliferation of neonatal rat cardiac fibroblasts.     

负载抗原肽的树突状细胞增强腺病毒载体介导hTRP2诱发抗小鼠黑色素瘤免疫

目的探讨人黑色素瘤相关抗原TRP2(hTRP2)MHCⅠ多肽TRP2p180-188 (SVYDFFVWL)修饰小鼠骨髓来源的树突状细胞(DC),增强腺病毒载体介导hTRP2(Ad hTRP2)诱发的抗小鼠黑色素瘤免疫效果。方法Ad hTRP2免疫C57BL／6小鼠,3周后分别用Ad hTRP2或DC／SVYDFFVWL再免疫一次。用体内细胞毒性T淋巴细胞(CTL)杀伤实验和细胞内IFNγ染色(ICS)分析CTL杀伤活性和IFNγ的产生;或给免疫小鼠皮下接种B16．F10细胞,观察荷瘤小鼠的成活情况。结果测定脾脏中6 h CTL杀伤率和产生IFNγ的CD8+T细胞比例,结果分别是:Ad hTRP2(初免)- Ad hTRP2(加强)组为68．40％±5．50％和0．67％±0．16％,DC／SVYDFFVWL(初免)-DC／SVYDFFVWL(加强)组为28．50％±6．40％和0．22％±0．07％,而Ad hTRP2(初免)-DC／SVYDFFVWL(加强)组为98．90％±0．90％和1．05％±0．21％。荷瘤实验显示,小鼠接种1×106 B16．F10细胞后3个月,DC／SVYDFFVWL(初免)-DC／SVYDFFVWL(加强)组小鼠无一成活,Ad hTRP2(初免)-Ad hTRP2(加强)组只有40％的小鼠存活,而Ad hTRP2(初免)-DC／SVYDFFVWL(加强)组小鼠100％存活。结论用DC／SVYDFFVWL加强免疫能显著地增强Ad hTRP2初次免疫的效果,表明Ad hTRP2(初免)-DC／SVYDFFVWL(加强)免疫是一种克服重复应用腺病毒并不能提高抗肿瘤免疫效应的有效方式。     

Preparation and properties of selected Zn(II)-peptide complexes in suspension

The preparation and properties of low soluble, suspended Zn(II) complexes containing the selected peptides: tyroliberin (TRH), gonadorelin (GnRH), dalarelin and corticothropin (ACTH) were studied. The amount of Zn(II) bound by 1 muM of the selected peptide (n) was defined, as well as affinity of Zn(II) to the peptide (Ka) and the durability of the created complex Zn(II)-peptide (Kd). ACTH associated the highest amount of Zn(II), and GnRH the lowest one: 1 microM of ACTH complexed 0.81 microM +/- 0.03 Zn(II), the same quantity of GnRH-0.52 microM +/- 0.07 and TRH and dalarelin associated 0.75+/-0.03 and 0.79+/-0.02 microM of Zn(II), respectively. The closest affinity was stated between Zn(II) and GnRH (Ka=157.692+/-21.300 microM(-1)), the smallest-towards ACTH (Ka=1.136+/-0.042 microM(-1)). The lower amount of Zn(II) associated by the studied peptide, the higher was its affinity versus this metal (r=-0.942). The analysis of the kinetics of the Zn(II)-peptide linkage revealed that the most stable complexes with this metal were formed by GnRH (Kd=0.006+/-0.001 microM(-1)) and by dalarelin (Kd=0.020+/-0.001 microM(-1)). Zn(II) with GnRH complexes are about 147 times more durable than ACTH (Kd=0.880+/-0.033 microM(-1)) ones. It was established that the Zn(II)-peptide complexes were more stable in the case of lower molecular weight of the peptide (r=0.963), and the inferior number of the amino acid residues accessible in the peptide (r=0.967).     

巨细胞病毒的形态学及结构多肽分析

对人巨细胞病毒的形态进行了观察,并对其结构多肽进行了SDS-聚丙烯酰胺凝胶电泳,结果表明巨细胞病毒有23条结构多肽。分子量范围19k～112k。     

The use of preparations of urinary bladder smooth muscle for bioassay of and discrimination between polypeptides

Summary Eleven polypeptides, two prostaglandins and three amines were assayed, in parallel, by measurement of their spasmogenic effect on the isolated urinary bladder of eight animal species, the in situ bladder of three species and the isolated ureter of three species. Several of the smooth muscle preparations examined proved to be sensitive and suitable test-objects for the quantitative bioassay of different peptides. At the same time they appeared to be useful for discriminating not only between peptides belonging to different groups, but also between members of the same peptide family. It is tentatively suggested that biogenic peptides may interfere in the physiological control of motility and tone of the urinary tract smooth muscle.Supported by grants from the Consiglio Nazionale delle Ricerche, Roma.     

Behavioural and electrocortical modifications induced in the rat by intraventricular injection of physalaemin and eledoisin

The effects of intraventricular injection of eledoisin and physalaemin in the rat were studied. Both peptides reduced explorative behaviour; eledoisin, but not physalaemin, produced catatonia, reduction of muscular strength, tremor and sialorrhea. The electrocorticogram showed the signs of a depressive effect both after physalaemin and eledoisin administration.     

Regulation der Azinuszellrezeptoren des Pankreas durch Peptide

Summary Peptides may act on the same receptor they regulate or on another receptor by causing regulations via receptor interactions. These receptor regulations include changes of receptor affinity and capacity. Receptor capacity is regulated by internalization, recycling, degradation, synthesis, and modification of bioavailability without migration of the receptor. Examples for those regulations, mostly based on experiments with isolated pancreatic acini from the rat, mouse, or guinea pig, are given.For the CCK receptor these examples include complex regulations of this receptor by CCK itself, bringing into discussion the hypothesis of negative cooperativity and the two-site receptor model, desensitization of the receptor by CCK, in vivo CCK influences on its receptor, and insulin receptor/CCK receptor interactions. For the insulin receptor the physiological significance of up and down regulation of this receptor by insulin itself is discussed. For the IGF receptors and the EGF receptor CCK-induced, Ca²⁺-mediated regulation of receptor internalization are another type of regulation with unknown physiological and pathophysiological significance. Finally CCK-induced, Ca²⁺-mediated regulation of somatostatin receptor capacity and affinity are mentioned.It is postulated that those regulations play an important role in influencing the biological effect of hormones and that knowledge about them may improve our understanding of pathophysiology.

---

Abkürzungsverzeichnis CCK Cholecystokinin - IGF insulin like growth factor - mRNA messenger-Ribonukleinsäure - MW Molekulargewicht - VIP vasoactive intestinal peptide     

Neuropeptide receptors in developing and adult rat spinal cord: An in vitro quantitative autoradiography study of calcitonin gene-related peptide,neurokinins, μ-opioid,galanin, somatostatin,neurotensin and vasoactive intestinal polypeptide receptors

A number of neuroactive peptides including calcitonin gene-related peptide (CGRP), substance P, neurokinin B, opioids, somatostatin (SRIF), galanin, neurotensin and vasoactive intestinal polypeptide (VIP) have been localized in adult rat spinal cord and are considered to participate either directly and/or indirectly in the processing of sensory, motor and autonomic functions. Most of these peptides appear early during development, leading to the suggestion that peptides, in addition to their neurotransmitter/neuromodulator roles, may possibly be involved in the normal growth and maturation of the spinal cord. To provide an anatomical substrate for a better understanding of the possible roles of peptides in the ontogenic development of the cord, we investigated the topographical profile as well as variation in densities of [¹²⁵I]hCGRPα, [¹²⁵I]substance P/neurokinin-1 (NK-1), [¹²⁵I]eledoisin/neurokinin-3 (NK-3), [¹²⁵I]FK 33–824 ([D-Ala², Me-Phe⁴, Met(O)ol⁵]enkephalin)/μ-opioid, [¹²⁵I]galanin, [¹²⁵I]T₀D₈-SRIF₁₄ (an analog of sornatostatin), [¹²⁵I]neurotensin and [¹²⁵I]VIP binding sites in postnatal and adult rat spinal cord using in vitro quantitative receptor autoradiography. Receptor binding sites recognized by each radioligand are found to be distributed widely during early stages of postnatal development and then to undergo selective modification to attain their adult profile of distribution during the third week of postnatal development. The apparent density of various receptor sites, however, are differently regulated depending on the lamina and the stage of development studied. For example, the density of μ-opioid binding sites, following a peak at postnatal day 4 (N), declines gradually in almost all regions of the spinal cord with the increasing age of the animal [¹²⁵I]substance P/NK-1 binding sites, on the other hand, show very little variation until P14 and then subsequently decrease as the development proceeds. In the adult rat, most of these peptide receptor binding sites are localized in relatively high amounts in the superficial laminae of the dorsal horn. To varying extents, moderate to low density of various peptide receptor binding sites are also found to be present in the ventral horn, intermediolateral cell column and around the central canal. Taken together, these results suggest that each receptor-ligand system is regulated differently during development and may each uniquely be involved in cellular growth, differentiation and in maturation of the normal neural circuits of the spinal cord, Furthermore, the selective localization of various receptor binding sites in adult rat spinal cord over a wide variety of functionally distinct regions reinforces the neurotransmitter/ modulator roles of these peptides in sensory, motor and autonomic functions associated with the spinal cord. © 1995 Wiley-Liss, Inc.     

Midbrain central gray: LHRH infusion enhances lordotic behavior in estrogen-primed ovariectomized rats

Ovariectomized rats were implanted with 23 gauge stainless steel cannulae in the ventrolateral midbrain central gray. Twelve sexually active rats were estrone-primed and infused with saline and 50 ng luteinizing hormone-releasing hormone (LHRH) in counterbalanced order. Infusion of luteinizing hormone-releasing hormone significantly enhanced the lordotic response to coital stimulation compared to saline infusion. These results support the role of hypothalamo-mesencephalic LHRH-containing pathways in modulating lordotic behavior in estrogen-primed Ovariectomized rats.