清醒大鼠室旁核灌流谷氨酸受体阻断剂对压力感受性反射的影响

目的:探讨下丘脑室旁核(PVN)内谷氨酸参与压力感受性反射中枢调节的神经化学机制。方法:在清醒大鼠,用脑部微量透析法和高效液相色谱法观察静脉注射苯肾上腺素诱发压力感受性反射对PVN区谷氨酸含量的影响;NMDA受体阻断剂MK-801或非NMDA受体阻断剂CNQX直接灌流PVN区并诱发压力感受性反射,进一步探讨PVN区谷氨酸对压力感受性反射的作用。结果:①静脉注射苯肾上腺素诱发压力感受性反射时,PVN内的谷氨酸含量迅速升高到注射前的384.82%±91.77%(P<0.01)。②PVN区灌流谷氨酸受体阻断剂MK-801或CNQX,同时诱发压力感受性反射,其血压升值明显减少,心率降值明显增加(P<0.01),压力感受性反射的敏感性(△HR/△MAP)明显增加(P<0.01)。结论:PVN内的谷氨酸可能通过离子型谷氨酸受体参与压力感受性反射的中枢调节,而且此调节作用可能是抑制性的。     

中枢血管紧张素对心血管活动调节作用

血管紧张素(Ang)广泛存在于中枢神经系统和外周组织中,对心血管活动和交感神经活动起重要调节作用。本文介绍了孤束核(NTS)、延髓头端腹侧区(RVLM)、延髓尾端腹侧区(CVLM)和室旁核(PVN)内Ang对心血管活动的影响,Ang对动脉压力感受性反射(ABR)和心交感传入反射(CSAR)的调节作用,肾素-血管紧张素系统的基因敲除研究,以及Ang与高血压和慢性心力衰竭的关系。     

Genistein对大鼠垂体前叶细胞增殖的抑制作用

应用细胞培养、^3H-TdR掺入、流式细胞和电镜技术,观察酪氨酸蛋白激酶（PTK）抑制剂genistein对正常大鼠垂体前叶细胞和垂体瘤细胞株AtT-20增殖的影响,并探讨其可能的机制。结果显示：genistein作用48h后可明显抑制正常大鼠垂体前叶细胞和垂体瘤细胞株AtT-20增殖。流式细胞仪检测发现,50和100μmol/L genistein可将AtT-20细胞阻断于G0／G1期及G2／M期,并出现凋亡峰,凋亡率分别灰19.9％和36.4％。电镜照片显示有凋亡细胞。结果表明,PTK抑制剂可以明显抑制正常大鼠垂体前叶细胞和垂体瘤细胞株AtT-20的殖,并诱导细胞凋亡,说明PTK活性对细胞增殖和分化有重要作用。     

孤束核组胺受体参与大鼠脑室注射组胺对颈动脉窦反射的抑制

目的:探讨孤束核(NTS)组胺(HA)受体在脑室注射(ICV) HA对颈动脉窦压力感受性反射(CBR)影响中的作用.方法:分离麻醉大鼠的双侧颈动脉窦区,将不同窦内压(ISP)与其对应的平均动脉压(MAP)值进行Logistic五参数曲线拟合,求得ISP-MAP关系曲线及其特征参数,观察ICV HA以及预先在NTS微量注射HA受体拮抗剂对CBR的作用.结果:ICV HA (100 ng)导致ISP-MAP关系曲线显著上移,ISP和增益关系曲线中部明显下移,反射参数中阈压、饱和压和最大增益时的窦内压值增大,MAP反射变动范围及反射最大增益减小.预先向NTS内注射H1或H2受体拮抗剂氯苯吡胺(CHL,0.5 μg)或西咪替丁(CIM,1.5 μg),均可明显减弱HA的上述效应,CIM的这种减弱作用不如CHL的显著.结论:脑室给HA使CBR产生快速重调定,反射敏感性下降,NTS的H1和H2受体尤以H1受体在ICV HA抑制CBR的机制中发挥重要作用,下丘脑-NTS的HA能通路可能是HA调节CBR的下行通路之一.     

PTK重组质粒的构建与表达

目的构建并表达出PTK重组蛋白,以鉴定酪氨酸蛋白激酶(PTK)的活性,筛选酪氨酸激酶的抑制剂。方法从PTK重组克隆载体上切下目的基因片段Abl-PTK使其连接到表达载体pGEX4T-2上,转化大肠杆菌DH5a,筛选鉴定出正确的转化子。转化菌株经IPTG诱导后进行表达并进行SDS-PAGE分析。结果经酶切和诱导表达鉴定,重组质粒pGEX4T-2-PTK构建成功,并高效表达了58KD的GST-PTK融合蛋白。结论PTK基因被成功地重组至融合蛋白表达载体中,并在大肠杆菌中获得高效表达。该研究为进一步纯化、鉴定PTK的活性,筛选PTK的抑制剂奠定了基础。     

家兔脑内微量注射或脊髓珠网膜下腔注射氟安定的呼吸抑制作用

实验在49只局部麻醉、肌肉麻痹、切断双侧颈迷走神经的家兔上进行。观察到一侧孤束核(NTS)区微量注射氟安定,使对侧膈神经放电平均幅度减低,呼吸频率加快。事先在NTS区微量注射GABA 受体拮抗剂印防己毒素,可阻断氟安定的减低膈神经放电平均幅度的作用。在脑桥头端横断脑干出现长吸式呼吸的免上,一侧NTS区微量注射氟安定,也使对侧膈神经放电平均幅度减低,但呼吸频率减慢。在脑桥结合臂旁内侧核(NPBM)区微量注射氟安定,使呼吸频率明显减慢,吸气和呼气时程延长,这效应可被毒扁豆碱阻断。脊髓蛛网膜下腔注射氟安定,使膈神经放电平均幅度减低,血压下降,这效应可被印防己毒素阻断。结果提示,NPBM 的存在与呼吸频率加快有关;而氟安定作用于NPBM区,却减慢呼吸频率,其机制可能是抗胆碱能作用。氟安定直接作用于NTS 区或脊髓使膈神经放电平均幅度减低、血压下降,其机制可能与激活GABA 受体有关。     

猫颈动脉区压力和化学感受性刺激对孤束核自发放电的影响

对27只氯醛糖和氨基甲酸乙酯麻醉的猫观察了选择性激活颈动脉压力和化学感受器对孤束核(NTS)及其附近区域单位放电的影响。共记录到103个对颈动脉压力感受器激活(新福林1—2μg/kg,iv)和/或颈动脉化学感受器激活(尼古丁,5—20μg,注入甲状腺动脉)起反应的单位,其中81个位于NTS,在这些单位中,14个仅对化学感受器激活起反应(10个兴奋和4个抑制),44个以各种组合形式对化学和压力感受性刺激都起反应,23个仅对压力感受器刺激起反应(18个兴奋和5个抑制)。在定位分布上,对两种刺激都起反应的单位主要位于NTS尾部,仅对化学感受性刺激起反应的单位多位于NTS的腹外侧,其它有反应的单位分别位于舌下神经旁区,旁正中网状核和延髓腹侧尾端。这些结果表明,颈动脉区压力和化学感受器活动传入到NTS,并在其中的一些神经元上发生会聚。 在全部有反应的NTS单位中,68个对压力感受性刺激起反应,其中46个兴奋,21个抑制(P<0.005);58个对化学感受性刺激起反应,其中36个兴奋,22个抑制(P>0.05)。这些结果提示,化学感受性刺激对NTS神经元引起兴奋和抑制两种反应,而压力感受性刺激则诱发兴奋为主的反应。     

兔脑内微量注射纳洛酮和横断脑干对吗啡呼吸抑制效应的影响

本工作用静脉和脑内微量注射吗啡与纳洛酮,以及横断脑干等方法,分析了静脉内注射吗啡引起呼吸抑制效应的机制。实验在53只家兔上进行。结果观察到,在一侧脑桥结合臂旁内侧核区(NPBM)微量注射吗啡(20μg/2μl),可使双侧膈神经放电抑制,其抑制规律与静脉内注射吗啡(3mg/kg)的规律相似。当静脉注射吗啡造成呼吸抑制后,于一侧 NPBM 区微量注射纳洛酮(4μg/2μl)或横断脑干部分去除 NPBM,均可使膈神经放电产生脱抑制。横断脑干去除 NPBM 后,在长吸状态下吗啡呼吸抑制效应不易出现;如进一步在髓纹前方1—2mm处横断脑干,则静脉注射吗啡又能引起膈神经放电抑制。在一侧孤束核区(NTS)微量注射吗啡,可引起对侧膈神经放电抑制,而同侧不抑制。上述结果提示,静脉内注射吗啡引起的呼吸抑制效应,主要是通过 NPBM 活动改变而实现的,但 NTS 活动改变在其中也起一定作用;而“长吸中枢”的活动则阻碍吗啡呼吸抑制效应的出现。     

甲壳胺对HepG2细胞蛋白酪氨酸激酶和磷酸酶活性的影响

目的:初步探讨甲壳胺诱导人肝癌Hep G2细胞凋亡的信号转导机制。方法:采用酶联免疫法,动态检测甲壳胺作用于Hep G2细胞后,细胞膜相及胞浆内的蛋白酪氨酸激酶(PTK)及蛋白酪氨酸磷酸酶(PTP)活性的变化。结果:甲壳胺可以抑制Hep G2细胞内的PTK活性,并呈一定的浓度依赖性;甲壳胺作用Hep G2细胞后,随着PTK活性的减弱,PTP的活性也短暂下降。结论:甲壳胺诱导Hep G2细胞凋亡时,涉及到PTK的活性改变。观察到膜相蛋白中PTK的活性改变早于胞浆蛋白,提示可能存在一个信号的跨膜转运过程;同时伴有PTP的活性变化,可能反映了胞内蛋白酪氨酸残基的磷酸化与去磷酸化即时调节机制。     

蛋白激酶C和蛋白酪氨酸激酶介导脂多糖及细胞因子诱导大鼠血管平滑肌细胞一氧化氮的生成

采用Spprague-Dawley大鼠胸主动脉中膜、外膜和培养的血管平滑肌细胞(VSMCs)作材料,鉴定不同类型的血管组织经炎性介质刺激后其一氧化氮(NO)的产生来源,闻明蛋白激酶C(PKC)和蛋白酪氨酸激酶(PTK)介导大鼠VSMCs生成NO的调控机制。大鼠VSMCs经脂多糖(LPG)和细胞因子(TNF-α,IL-1β)处理后,以剂量依赖方式促进NO释放。采用Western Blot证实经刺激的VSMCs伴有iNOS表达上调。进一步实验表明PKC和PTK参与LPS和细胞因子诱导NO生成的胞内信号转导。用PKC抑制剂H7与VSMCs共培育,H7能明显减少LPS、TNF-α和IL-1β诱导细胞NO的形成。白屈菜赤碱亦可抑制NO的生成,但HAl004对VSMCs的NO生成无抑制作用,提示PKC参与NO的生成与调控。PTK抑制剂genistein和tyrphostin AG18均能抑制由LPS、TNF-α和IL-1β引发VSMCs释放NO,同时伴iNOS蛋白表达下调,而PKC抑制剂不能阻断iNOS的表达。上述观察结果提示,PKC介导LPS和细胞因子诱导细胞合成NO可能是通过iNOS翻译后加工;而PTK则以上调iNOS表达而促增NO生成。     

Chronic hypoxia abolishes posthypoxia frequency decline in the anesthetized rat

In anesthetized rats, increases in phrenic nerve amplitude and frequency during brief periods of hypoxia are followed by a reduction in phrenic nerve burst frequency [posthypoxia frequency decline (PHFD)]. We investigated the effects of chronic exposure to hypoxia on PHFD and on peripheral and central O2-sensing mechanisms. In Inactin-anesthetized (100 mg/kg) Sprague-Dawley rats, phrenic nerve discharge and arterial pressure responses to 10 s N2 inhalation were recorded after exposure to hypoxia (10 +/- 0.5% O2) for 6-14 days. Compared with rats maintained at normoxia, PHFD was abolished in chronic hypoxic rats. Because of inhibition of PHFD, the increased phrenic burst frequency and amplitude after N2 inhalation persisted for 1.8-2.8 times longer in chronic hypoxic (70 s) compared with normoxic (25-40 s) rats (P < 0.05). After acute bilateral carotid body denervation, N2 inhalation produced a short depression of phrenic nerve discharge in both chronic hypoxic and normoxic rats. However, the degree and duration of depression of phrenic nerve discharge was smaller in chronic hypoxic compared with normoxic rats (P < 0.05). We conclude that after exposure to chronic hypoxia, a reduction in PHFD contributes to an increased duration of the acute hypoxic ventilatory response in anesthetized rats. Furthermore, after exposure to chronic hypoxia, the central network responsible for respiration is more resistant to the depressant effects of acute hypoxia in anesthetized rats.     

Effects of enoximone on peripheral and central chemoreflex responses in humans

cAMP plays an important role in peripheral chemoreflex function in animals. We tested the hypothesis that the phosphodiesterase inhibitor and inotropic medication enoximone increases peripheral chemoreflex function in humans. In a single-blind, randomized, placebo-controlled crossover study of 15 men, we measured ventilatory, muscle sympathetic nerve activity, and hemodynamic responses to 5 min of isocapnic hypoxia, 5 min of hyperoxic hypercapnia, and 3 min of isometric handgrip exercise, separated by 1 wk, with enoximone and placebo administration. Enoximone increased cardiac output by 120 +/- 3.7% from baseline (P < 0.001); it also increased the ventilatory response to acute hypoxia [13.6 +/- 1 vs. 11.2 +/- 0.7 l/min at 5 min of hypoxia, P = 0.03 vs. placebo (by ANOVA)]. Despite a larger minute ventilation and a smaller decrease in O(2) desaturation (83 +/- 1 vs. 79 +/- 2%, P = 0.003), the muscle sympathetic nerve response to hypoxia was similar between enoximone and placebo (123 +/- 6 and 117 +/- 6%, respectively, P = 0.28). In multivariate regression analyses, enoximone enhanced the ventilatory (P < 0.001) and sympathetic responses to isocapnic hypoxia. Hyperoxic hypercapnia and isometric handgrip responses were not different between enoximone and placebo (P = 0.13). Enoximone increases modestly the chemoreflex responses to isocapnic hypoxia. Moreover, this effect is specific for the peripheral chemoreflex, inasmuch as central chemoreflex and isometric handgrip responses were not altered by enoximone.     

Glucocorticoids reduce responses to AMPA receptor activation and blockade in nucleus tractus solitarius

We tested the hypothesis that glucocorticoids attenuate changes in arterial pressure and renal sympathetic nerve activity (RSNA) in response to activation and blockade of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) receptors within the nucleus of the solitary tract (NTS). Experiments were performed in Inactin-anesthetized male Sprague-Dawley rats treated for 7 +/- 1 days with a subcutaneous corticosterone (Cort) pellet or in control rats. Baseline mean arterial pressure (MAP) was significantly higher in Cort-treated rats (109 +/- 2 mmHg, n = 39) than in control rats (101 +/- 1 mmHg, n = 48, P < 0.05). In control rats, microinjection of AMPA (0.03, 0.1, and 0.3 pmol/100 nl) into the NTS significantly decreased MAP at all doses and decreased RSNA at 0.1 and 0.3 pmol/100 nl. Responses to AMPA in Cort-treated rats were attenuated at all doses of AMPA (P < 0.05). Responses to the AMPA-kainate receptor antagonist 6-cyano-7-nitroquinoxaline-2,3-dione (CNQX) were also significantly reduced in Cort-treated rats relative to control rats. Blockade of glucocorticoid type II receptors with mifepristone significantly enhanced responses to CNQX in both control and Cort rats. We conclude that glucocorticoids attenuate MAP and RSNA responses to activation and blockade of AMPA receptors in the NTS.     

Differential effects of tyrosine kinase inhibitors on volume-sensitive chloride current in human atrial myocytes: evidence for dual regulation by Src and EGFR kinases

To determine whether protein tyrosine kinase (PTK) modulates volume-sensitive chloride current (I(Cl.vol)) in human atrial myocytes and to identify the PTKs involved, we studied the effects of broad-spectrum and selective PTK inhibitors and the protein tyrosine phosphatase (PTP) inhibitor orthovanadate (VO(4)(-3)). I(Cl.vol) evoked by hyposmotic bath solution (0.6-times isosmotic, 0.6T) was enhanced by genistein, a broad-spectrum PTK inhibitor, in a concentration-dependent manner (EC(50) = 22.4 microM); 100 microM genistein stimulated I(Cl.vol) by 122.4 +/- 10.6%. The genistein-stimulated current was inhibited by DIDS (4,4'-diisothiocyanostilbene-2,2'-disulfonic acid, 150 microM) and tamoxifen (20 microM), blockers of I(Cl.vol). Moreover, the current augmented by genistein was volume dependent; it was abolished by hyperosmotic shrinkage in 1.4T, and genistein did not activate Cl(-) current in 1T. In contrast to the stimulatory effects of genistein, 100 microM tyrphostin A23 (AG 18) and A25 (AG 82) inhibited I(Cl.vol) by 38.2 +/- 4.9% and 40.9 +/- 3.4%, respectively. The inactive analogs, daidzein and tyrphostin A63 (AG 43), did not alter I(Cl.vol). In addition, the PTP inhibitor VO(4)(-3) (1 mM) reduced I(Cl.vol) by 53.5 +/- 4.5% (IC(50) = 249.6 microM). Pretreatment with VO(4)(-3) antagonized genistein-induced augmentation and A23- or A25-induced suppression of I(Cl.vol). Furthermore, the selective Src-family PTK inhibitor PP2 (5 microM) stimulated I(Cl.vol), mimicking genistein, whereas the selective EGFR (ErbB-1) kinase inhibitor tyrphostin B56 (AG 556, 25 microM) reduced I(Cl.vol), mimicking A23 and A25. The effects of both PP2 and B56 also were substantially antagonized by pretreatment with VO(4)(-3). The results suggest that I(Cl.vol) is regulated in part by the balance between PTK and PTP activity. Regulation is complex, however. Src and EGFR kinases, distinct soluble and receptor-mediated PTK families, have opposing effects on I(Cl.vol), and multiple target proteins are likely to be involved.     

Central nitric oxide modulates hindquarter vasodilation elicited by AMPA receptor stimulation in the NTS of conscious rats

Microinjection of S-alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionic acid (AMPA) in the nucleus of the solitary tract (NTS) of conscious rats causes hypertension, bradycardia, and vasoconstriction in the renal, mesenteric, and hindquarter vascular beds. In the hindquarter, the initial vasoconstriction is followed by vasodilation with AMPA doses >5 pmol/100 nl. To test the hypothesis that this vasodilation is caused by activation of a nitroxidergic pathway in the NTS, we examined the effect of pretreatment with the nitric oxide synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 10 nmol/100 nl, microinjected into the NTS) on changes in mean arterial pressure, heart rate, and regional vascular conductance (VC) induced by microinjection of AMPA (10 pmol/100 nl in the NTS) in conscious rats. AMPA increased hindquarter VC by 18 +/- 4%, but after pretreatment with L-NAME, AMPA reduced hindquarter VC by 16 +/- 7% and 17 +/- 9% (5 and 15 min after pretreatment, P < 0.05 compared with before pretreatment). Pretreatment with L-NAME reduced AMPA-induced bradycardia from 122 +/- 40 to 92 +/- 32 beats/min but did not alter the hypertension induced by AMPA (35 +/- 5 mmHg before pretreatment, 43 +/- 6 mmHg after pretreatment). Control injections with D-NAME did not affect resting values or the response to AMPA. The present study shows that stimulation of AMPA receptors in the NTS activates both vasodilatatory and vasoconstrictor mechanisms and that the vasodilatatory mechanism depends on production of nitric oxide in the NTS.     

Chronic hypoxia enhances the phrenic nerve response to arterial chemoreceptor stimulation in anesthetized rats.

Chronic exposure to hypoxia results in a time-dependent increase in ventilation called ventilatory acclimatization to hypoxia. Increased O(2) sensitivity of arterial chemoreceptors contributes to ventilatory acclimatization to hypoxia, but other mechanisms have also been hypothesized. We designed this experiment to determine whether central nervous system processing of peripheral chemoreceptor input is affected by chronic hypoxic exposure. The carotid sinus nerve was stimulated supramaximally at different frequencies (0.5-20 Hz, 0.2-ms duration) during recording of phrenic nerve activity in two groups of anesthetized, ventilated, vagotomized rats. In the chronically hypoxic group (7 days at 80 Torr inspired PO(2)), phrenic burst frequency (f(R), bursts/min) was significantly higher than in the normoxic control group with carotid sinus nerve stimulation frequencies >5 Hz. In the chronically hypoxic group, peak amplitude of integrated phrenic nerve activity ( integral Phr, percent baseline) or change in integral Phr was significantly greater at stimulation frequencies between 5 and 17 Hz, and minute phrenic activity ( integral Phr x f(R)) was significantly greater at stimulation frequencies >5 Hz. These experiments show that chronic hypoxia facilitates the translation of arterial chemoreceptor afferent input to ventilatory efferent output through a mechanism in the central nervous system.     

Differential role of PTK and ERK MAPK in superoxide impairment of K(ATP) and K(Ca) channel cerebrovasodilation

Previously, superoxide (O2 -) has been observed to impair pial artery dilation (PAD) to activators of the ATP-sensitive (KATP) and calcium-sensitive (KCa) K+ channels. This study tested the hypothesis that activation of protein tyrosine kinase (PTK) and the ERK isoform of MAPK by O2 - contribute to impairment of KATP and KCa channel PAD. Exposure of the cerebral cortex to a xanthine oxidase O2 --generating system (OX) blunted PAD to cromakalim, a KATP agonist, but preadministration of genistein, a PTK antagonist, or U-0126, an ERK MAPK inhibitor, almost completely prevented such impairment (11 +/- 1 and 22 +/- 1 vs. 3 +/- 1 and 7 +/- 1 vs. 10 +/- 1 and 16 +/- 2% for cromakalim with 10-8 and 10-6 M PAD during control, OX, and OX + genistein conditions). In contrast, neither genistein nor U-0126 robustly protected PAD to NS-1619, a KCa agonist, after OX exposure (11 +/- 1 and 18 +/- 2 vs. 1 +/- 1 and 2 +/- 1 vs. 4 +/- 1 and 6 +/- 1% for 10-8 and 10-6 M NS-1619 during control, OX, and OX + genistein conditions). These data show that PTK and ERK MAPK activation contribute to O2 --induced KATP and KCa channel PAD impairment and suggest a differential greater role for PTK and ERK MAPK in KATP vs. KCa channel PAD impairment.     

Chronic intermittent hypoxia alters NMDA and AMPA-evoked currents in NTS neurons receiving carotid body chemoreceptor inputs

Chronic exposure to intermittent hypoxia (CIH) has been used in animals to mimic the arterial hypoxemia that accompanies sleep apnea. Humans with sleep apnea and animals exposed to CIH have elevated blood pressures and augmented sympathetic nervous system responses to acute exposures to hypoxia. To test the hypothesis that exposure to CIH alters neurons within the nucleus of the solitary tract (NTS) that integrate arterial chemoreceptor afferent inputs, we measured whole cell currents induced by activation of alpha-amino-3-hydroxy-5-methyl-4-isoxazolepropionate (AMPA) and N-methyl-D-aspartate (NMDA) receptors in enzymatically dispersed NTS neurons from normoxic (NORM) and CIH-exposed rats (alternating cycles of 3 min at 10% O2 followed by 3 min at 21% O2 between 8 AM and 4 PM for 7 days). To identify NTS neurons receiving carotid body afferent inputs the anterograde tracer 4- (4-(dihexadecylamino)styryl-N-methylpyridinum iodide (DiA) was placed onto the carotid body 1 wk before exposure to CIH. AMPA dose-response curves had similar EC50 but maximal responses increased in neurons isolated from DiA-labeled CIH (20.1 +/- 0.8 microM, n = 9) compared with NORM (6.0 +/- 0.3 microM, n = 8) rats. NMDA dose-response curves also had similar EC50 but maximal responses decreased in CIH (8.4 +/- 0.4 microM, n = 8) compared with NORM (19.4 +/- 0.6 microM, n = 9) rats. These results suggest reciprocal changes in the number and/or conductance characteristics of AMPA and NMDA receptors. Enhanced responses to AMPA receptor activation could contribute to enhanced chemoreflex responses observed in animals exposed to CIH and humans with sleep apnea.     

Catestatin, a chromogranin A-derived peptide, is sympathoinhibitory and attenuates sympathetic barosensitivity and the chemoreflex in rat CVLM

Hypertension is a major cause of morbidity. The neuropeptide catestatin [human chromogranin A-(352-372)] is a peptide product of the vesicular protein chromogranin A. Studies in the periphery and in vitro studies show that catestatin blocks nicotine-stimulated catecholamine release and interacts with β-adrenoceptors and histamine receptors. Catestatin immunoreactivity is present in the rostral ventrolateral medulla (RVLM), a key site for blood pressure control in the brain stem. Recently, we reported that microinjection of catestatin into the RVLM is sympathoexcitatory and increases barosensitivity. Here, we report the effects of microinjection of catestatin (1 mM, 50 nl) into the caudal ventrolateral medulla (CVLM) in urethane-anesthetized, bilaterally vagotomized, artificially ventilated Sprague-Dawley rats (n = 8). We recorded resting arterial pressure, splanchnic sympathetic nerve activity, phrenic nerve activity, heart rate, and measured cardiovascular homeostatic reflexes. Homeostatic reflexes were evaluated by measuring cardiovascular responses to carotid baroreceptor and peripheral chemoreceptor activation. Catestatin decreased basal levels of arterial pressure (-23 ± 4 mmHg), sympathetic nerve activity (-26.6 ± 5.7%), heart rate (-19 ± 5 bpm), and phrenic nerve amplitude (-16.8 ± 3.3%). Catestatin caused a 15% decrease in phrenic inspiratory period (T(i)) and a 16% increase in phrenic expiratory period (T(e)) but had no net effect on the phrenic interburst interval (T(tot)). Catestatin decreased sympathetic barosensitivity by 63.6% and attenuated the peripheral chemoreflex (sympathetic nerve response to brief hypoxia; range decreased 39.9%; slope decreased 30.1%). The results suggest that catestatin plays an important role in central cardiorespiratory control.     

Differential role of the paraventricular nucleus of the hypothalamus in modulating the sympathoexcitatory component of peripheral and central chemoreflexes

In the present study we investigated the involvement of the hypothalamic paraventricular nucleus (PVN) in the modulation of sympathoexcitatory reflex activated by peripheral and central chemoreceptors. We measured mean arterial blood pressure (MAP), heart rate (HR), renal sympathetic nerve activity (RSNA), and phrenic nerve activity (PNA) before and after blocking neurotransmission within the PVN by bilateral microinjection of 2% lidocaine (100 nl) during specific stimulation of peripheral chemoreceptors by potassium cyanide (KCN, 75 microg/kg iv, bolus dose) or stimulation of central chemoreceptors with hypercapnia (10% CO(2)). Typically stimulation of peripheral chemoreceptors evoked a reflex response characterized by an increase in MAP, RSNA, and PNA and a decrease in HR. Bilateral microinjection of 2% lidocaine into the PVN had no effect on basal sympathetic and cardiorespiratory variables; however, the RSNA and PNA responses evoked by peripheral chemoreceptor stimulation were attenuated (P < 0.05). Bilateral microinjection of bicuculline (50 pmol/50 nl, n = 5) into the PVN augmented the RSNA and PNA response to peripheral chemoreceptor stimulation (P < 0.05). Conversely, the GABA agonist muscimol (0.2 nmol/50 nl, n = 5) injected into the PVN attenuated these reflex responses (P < 0.05). Blocking neurotransmission within the PVN had no effect on the hypercapnia-induced central chemoreflex responses in carotid body denervated animals. These results suggest a selective role of the PVN in processing the sympathoexcitatory and ventilatory component of the peripheral, but not central, chemoreflex.