降钙素基因相关肽对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白表达及磷吸收的影响

本试验旨在研究不同浓度降钙素基因相关肽(CGRP)对猪小肠上皮细胞钠离子依赖型Ⅱb磷转运蛋白(NaPi-Ⅱb)表达和磷吸收的影响.选用第3～8代猪小肠上皮细胞进行试验,按单因素试验设计,将试验细胞分为1个对照组和5个CGRP(1×10-11～1×10-7 mmol/L)试验组,每组设6个重复.用4-甲基偶氮唑盐(MTT)法测定细胞生长速度,采用比色法测定CGRP干预24 h后细胞上清液磷含量,用RT-PCR法检测NaPi-Ⅱb mRNA相对表达量,以及蛋白质印迹(Western blot)法检测NaPi-Ⅱb蛋白表达.结果表明:与对照组相比,不同浓度CGRP对细胞生长没有显著影响(P＞0.05);各试验组均极显著或显著地促进了磷的吸收和NaPi-ⅡbmRNA的相对表达量(P＜0.01或P＜0.05);除1×10-10 mmol/L组外均显著提高了NaPi-Ⅱb蛋白的表达(P＜0.05).研究结果表明,CGRP通过诱导小肠上皮细胞中NaPi-Ⅱb的表达促进细胞磷的吸收,其中以1×10-9、1×10-8 mmol/L浓度的CGRP促进效果较为显著.     

Immunohistochemical analysis of pseudorabies virus spread through neurons innervating the eyeball

Pseudorabies virus (PRV) was inoculated intraocularly into BALB/c mice, and the distribution pattern of cells positive for several neurotransmitters and viral antigens in the eyeball, trigeminal nerve ganglia, and superior cervical ganglia was examined immunohistochemically to clarify the neural route of the virus spread. In the eyeball, substance P (SP)- and calcitonin gene-related peptide (CGRP)-positive cells were detected in the ipsilateral iris and ciliary body, neuropeptide tyrosine (NPY)-positive cells were detected in the choloid membrane, and tyrosine hydroxylase (TH)-positive cells were detected in the ipsilateral inner nuclear layer of the retina; all these cells contained viral antigens. In the superior cervical ganglia, viral antigen-positive cells containing TH or NPY were found at bilateral sites. In the trigeminal nerve ganglia, viral antigen-positive cells containing SP or CGRP were found at bilateral sites. These findings indicated that the SP- and CGRP-positive ganglion cells of the trigeminal nerve ganglia innervating the iris or ciliary body, and the NPY-positive ganglion cells of the superior cervical ganglia innervating the iris, ciliary body, and choroid membrane served as the route for the virus spread. These findings also suggested that dopaminergic neurons, such as the TH-positive retinal cells and TH-positive ganglion cells of the superior cervical ganglia, served as the route for virus spread.     

激活素A促进鸡胚背根神经节神经突起生长的作用机制

采用半定量PCR检测激活素II型受体（ActRII）、钙基因相关肽（cGRP）、血管活性肠肽（VIP）mRNA表达,液相一质谱联用仪检测5-羟色胺释放水平。结果显示,5μg／L激活素A能明显促进鸡胚背根神经节神经突起生长,同时可见激活素A促进ActRIIA、CGRPmRNA表达及神经递质5-羟色胺释放,但对ActRIIB及VIPmRNA表达无影响。结果表明,ActivinA可能通过ActivinA—ActRIIA途径发挥作用,其促进鸡胚背根神经节神经突起生长与其上调CGRP表达及5-HT释放有关。     

Transdermal behaviors comparisons among Evodia rutaecarpa extracts with different purity of evodiamine and rutaecarpine and the effect of topical formulation in vivo

Aim

Evodiamine (EVO) and rutaecapine (RUT), the major active components from Evodia rutaecarpa extract (EE), are recognized as a depended analgesic agent. This study was designed to investigate the effect of purity and chemical enhancers on the transdermal behavior of EVO and RUT, and the pharmacological effect of their topical cream in vivo.

Material and methods

Transdermal delivery across a full thickness pig abdominal skin was detected in vitro by Franz-type diffusion cell, with HPLC for quantification of the permeation of EVO and RUT. The activity of topical cream in vivo was evaluated by a mice pain model induced by formalin and hot plate.

Results

Transdermal characters of EVO and RUT showed a low transdermal rate, long lag time and low cumulative amount. The transdermal rate and cumulative amount could be promoted by lipophilic enhancers, whereas lag time was shortened by hydrophilic surfactant, but these permeation parameters were not markedly influenced by purity of EE (p > 0.05). The effect in vivo was confirmed by analgesic models in topical cream of EE, which produced a significant (p < 0.05) inhibitory effects on pain response in dose-dependent manner.

Conclusion

The purity of EVO and RUT from EE has no significant effect on their permeation through porcine skin, but oleic acid or nerolidol can markedly elevate the transdermal rate of EVO and RUT. High purity of EE is the best choice for topical preparation to increase the drug loading. The effect of EE in vivo is verified by formalin model and hot plate test.     

Localization of calcium regulatory hormones in fish

Immunocytochemical localization of hypocalcin, a hypocalcemic factor in the corpuscles of Stannius (CS), in American eels was examined at the light (ABC method) and electron microscopic (protein A-gold technique) levels with the specific antiserum raised against purified rainbow trout hypocalcin. Only type 1 cells in the CS were immunoreactive in the light microscopic immunocytochemistry. At the electron microscopic level, however, hypocalcin immunoreactivity was observed in secretory granules of both type 1 and type 2 cells. Our findings may indicate that type 1 cells are the main source of hypocalcin, but that type 2 cells also produce it, suggesting that the presence of two cell types reflects different physiological conditions of a single cell type, rather than functionally different cell types. In addition, we summarize our recent data on the localization of other calcium regulatory, or putative calcium regulatory, hormones in fish: parathyroid hormone, calcitonin and calcitonin gene-related peptide.     

降钙素基因相关肽在牦牛附睾中的定位与表达

【目的】分析降钙素基因相关肽（calcitonin gene related peptide,CGRP）在牦牛附睾中的转录水平及蛋白分布特点,探究其对精子运输及成熟的功能。【方法】采集6头3~4岁健康成年牦牛的附睾,将其解剖为头、体、尾三部分,应用实时荧光定量PCR（RT-qPCR）、Western blotting分别检测CGRP mRNA和蛋白在附睾的头、体、尾中的表达水平;采用H.E染色法分析附睾组织结构特点;采用免疫组化和免疫荧光技术分析附睾各部分CGRP的分布及表达情况。【结果】RT-qPCR结果表明,CGRP在牦牛附睾头的相对表达量显著高于附睾体和附睾尾（P＜0.05）,附睾尾表达量最低（P＜0.05）;Western blotting检测结果表明,CGRP蛋白水平在附睾尾中的表达水平显著高于附睾头和附睾体（P＜0.05）。显微镜下观察表明,牦牛附睾组织纤毛长度从头部至尾部逐渐降低,差异显著;附睾头间质组织的间质平滑肌层厚度显著低于附睾体和附睾尾（P＜0.05）;基细胞和晕细胞在附睾尾分布显著,上皮厚度亦在附睾尾显著降低。免疫组化及免疫荧光分析发现,CGRP主要分布于附睾上皮、基底膜及附睾间质小血管,附睾上皮晕细胞的表达强度最高。【结论】CGRP主要分布于牦牛附睾上皮和间质小血管,提示CGRP对牦牛附睾间质小血管的收缩发挥调节作用。牦牛附睾尾晕细胞数量最多,晕细胞中CGRP的高强度表达提示其可能参与附睾尾的免疫调节。     

小型猪复合麻醉剂对小型猪部分心肺指标及血浆内皮素和降钙素基因相关肽含量的影响

 【目的】研究小型猪复合麻醉剂对小型猪血浆内皮素(endothelin, ET)和降钙素基因相关肽(calcitonin gene related peptide, CGRP)含量及部分心肺指标的影响。【方法】14头实验用小型猪肌肉注射小型猪复合麻醉剂0.15 mL&#8226;kg-1,并在注药前及注药后5、10、30、45、60、80、100、120 min分别进行无创血压和心率(heart rate, HR)的监测并同步采取前腔静脉血样。采用放射性免疫分析法测定小型猪血浆中ET和CGRP的含量,并分析他们之间的相关性。【结果】无创血压、HR主要是在注药后5-10 min及80 min时变化明显(P＜0.01或P＜0.05)。ET与血压的变化趋势大致相似,且它们之间存在着一定的相关性(P＜0.05);CGRP与血压的变化趋势相反,呈一定的负相关性(P＜0.05)。【结论】ET和CGRP共同参与了小型猪复合麻醉剂引起的小型猪血流动力学变化过程,ET和CGRP的变化可能是小型猪复合麻醉及引起小型猪血压发生变化的主要原因之一。     

脾虚证模型大鼠血浆中β-EP、CGRP、NPY含量的变化

为了阐明脾虚证模型大鼠血浆中β-endorphin，β-EP）、降钙素基因相关肽（calctonin gene related peptide，CGRP）和神经肽Y（neuropeptide Y，NPY）变化规律和健脾益气方剂四君子汤对其的调节作用以及对脾虚证病理过程的保护机制，应用利血平复制出脾虚证大鼠模型，以特异性放射免疫分析法捡测了血浆β-EP、CGRP和NPY的含量变化。结果显示．脾虚证模型大鼠血浆β-EP、CGRP比对照组显著升高（P〈0．01）．NPY比对照组显著降低（P〈0．01）．接近对照组水平。提示脾虚证的病理变化过程与β-EP、CGRP、NPY等神经肽类的失衡有一定的相关性，健脾益气方剂四君子汤对脾虚证模型大鼠血浆内β-EP、CGRP和NPY含量变化有较好的调节作用，推测这也许是四君子汤健脾益气能够治疗脾虚证的作用机制之一。     

羊垂体前叶降钙素基因相关肽(CGRP)与P物质(SP)免疫反应神经纤维

应用免疫组化ＡＢＣ法发现羊垂体前叶有降钙素基因相关肽免疫反应神经纤维存在,这些纤维呈串球状分布于结节部及远侧部前２／３区域。纤维形态多样,细长者穿行于腺细胞之间;短小弯曲者围绕于腺细胞周围;反复分支折叠盘曲缠绕成团簇状者将腺细胞成堆的网络其中。神经纤维及串珠与腺细胞关系密切,提示有可能对腺细胞分泌起直接支配作用。在纤维分布区域可见散在的免疫反应细胞,形态不规则,大部分与神经纤维无接触关系,但有少数可位于纤维的网络之中。Ｐ物质免疫反应神经纤维及细胞在羊垂体前叶分布稀疏,且多位于结节部,远侧部很少。     

隆朋麻醉及其手术下对犬内皮素和降钙素基因相关肽的影响

探讨单纯隆朋麻醉和隆朋麻醉下手术对犬内皮素(ET)和降钙素基因相关肽(CGRP)分泌的影响。方法:选取健康犬12只,将其随机均分为单纯隆朋麻醉组和隆朋麻醉手术组。两组均于麻醉前、0.5 h、2 h、8 h、24 h、48 h、72 h和120 h采集静脉血,并迅速离心取血浆,用放射性免疫法检测ET和CGRP的浓度。结果:麻醉组,ET出现上升趋势而CGRP呈现下降趋势。ET到2 h时恢复至麻前水平,CGRP到8 h时恢复至麻前水平。手术组,手术开始后ET和CGRP都呈现上升趋势,两者都在24 h时恢复至正常水平。结论:隆朋单纯麻醉时对ET和CGRP有轻微的影响,但隆朋麻醉手术时ET和CGRP都出现上升趋势,机体的应激反应较强烈。