脑利钠肽BNP的临床应用进展

脑利钠肽(BNP)是利钠肽系统中的重要成员之一.BNP是一种包含了32个氨基酸的多肽,于1998年被日本学者从猪脑中发现并提取出来.它主要由心室肌细胞受压分泌,故可以反映左心室壁承受的压力大小.BNP可产生排钠利尿效应,参与调节水、电解质平衡,减少血浆容量;同时还可扩张血管,从而降低体循环血管阻力.有大量研究显示,血浆BNP水平在充血性心力衰竭中增高,因此它可作为心衰的有效监测指标之一,并且在临床上得到了广泛应用.此外,BNP还可作为早期心血管疾病的筛查高危因素.近年来BNP在诊疗心血管疾病、呼吸困难、肺栓塞、慢性阻塞性肺疾病和肺癌过程中均显示出重要作用.现就BNP在上述多学科领域中运用的新进展作一综述.     

N-terminal Pro-Brain Natriuretic Peptide And Atrial Fibrillation

NT-proBNP is produced from both atria and ventricles. The primary regulation of production is at the synthesis level. The plasma half-life of NT-proBNP is 60-120 min. Cutoff value of NT-proBNP for diagnosis of heart failure is 125 pg/ml in the age group below 75 years and 450 pg/ml in the age group above 75 years. It increases in atrial fibrillation and drops after successful cardioversion. High levels predict development of atrial fibrillation (AF) in healthy persons with sinus rhythm (SR). Some studies concluded that baseline level predicts maintenance of SR after cardioversion of AF while some others found that it did not. Many studies have proven that it is useful in monitoring rhythm stability after cardioversion of AF. Since it is increased in many other conditions, out of which some may also cause AF, care must be taken before ascribing changes in its level to AF alone.     

脑钠肽临床应用进展

脑钠肽(brain natriuretic peptide,BNP)是近年倍受关注的心血管生物标记物,BNP是一种主要由心脏分泌的肽类激素,在心脏维持其正常结构和功能的中起着重要的作用,它具有利钠、利尿、扩血管、降压、拮抗RAAS系统、抑制交感神经兴奋等作用.它已超过原来仅作为心衰的诊断检测指标范畴.研究表明BNP与呼吸困难的鉴别诊断、心肌梗死、高血压、心房颤动、心肌病、肺栓塞等关系密切,现就BNP的临床研究进展作一综述.     

Immunohistochemistry of Atrial Natriuretic Peptide in Brain Infarction

Atrial natriuretic peptide (ANP) was originally isolated from cardiac atria, and has potent natriuretic, diuretic, and vasorelaxant properties. It has been localized in neurons and astrocytes in the cerebral cortex and the white matter. We hypothesize that glial ANP may contribute to the regulation of cerebral blood flow in brain infarction. In order to elucidate this possible role, the immunohistochemistry of ANP was studied in cases of brain infarction and in other cases of brain trauma for comparison. A statistically significant increase in the number of ANP-immunoreactive glial cells (mainly astrocytes) was observed in the white matter surrounding the brain infarction compared with the intact area. No statistically significant increase in ANP-immunoreactive glial cell number was observed in the cerebral white matter from brain haemorrhage, contusion and control cases. Our results indicate that glial ANP may increase in number in brain infarction, and that it may be involved in the regulation of the cerebral blood flow in the infarcted area.     

Atrial Natriuretic Peptide Regulation of Vertebrate Intestinal Ion Transport

Atrial natriuretic peptide (ANP), in spite of its name, is synthesizedand secreted in vertebrates by both the atria and the ventriclesand also a number of extracardiac tissues. Likewise, the listof putative targets of ANP is large and includes, in additionto the kidney and vascular smooth muscle, the ion- and water-transportingintestine. Immunohistochemical staining of the intestine ofthe euryhaline marine goby Gillichthys mirabilis demonstratesthe presence of ANP-ergic neurons in the submucosa suggestingparacrine delivery to intestinal epithelial and smooth musclecells. ANP inhibits ion absorption across the goby intestine,supportingan osmoregulatory role for ANP.     

Expression of C-Type Natriuretic Peptide in the Bovine Pineal Gland

Abstract: The effect of lithium on inositol phospholipid resynthesis in primary cultures of cerebellar granule cells was studied. During activation of phospholipase C by the combined action of a muscarinic agonist and mild depolarization, the levels of inositol phospholipids as well as the inositol phospholipid precursor CMP-phosphatidate appeared highly sensitive to lithium with half-maximal accumulation of CMP-phosphatidate attained at 0.5 m M LiCl, a concentration close to that in the plasma of patients subjected to lithium therapy. Under the same conditions, the effect of lithium on inositol phospholipid metabolism appeared to be mediated by depletion of cytoplasmic free inositol content. This was indicated by the observation that preincubation for 48 h in high extracellular inositol concentrations could decrease or delay the depletion of inositol phospholipids and the accumulation of CMP-phosphatidate induced by 10 m M LiCl. Because even relatively high concentrations of extracellular inositol (500 µ M ) only partially prevented inositol phospholipid depletion, cerebellar granule cells appear to have a comparatively low capacity to accumulate inositol intracellularly, in comparison with other brain cells in culture. The relationship between CMP-phosphatidate accumulation and phospholipase C activity has also been investigated using a range of agonists that have been reported to act on cerebellar granule cells.     

人γ心钠素在大肠杆菌中的高效表达

利用聚合酶链式反应（ＰＣＲ）技术扩增人γ心钠素（γ－ｈＡＮＰ）ｃＤＮＡ编码序列,在其５′和３′端分别引入ＥｃｏＲⅠ和ＢａｍＨⅠ限制性内切酶位点,定向克隆到表达质粒载体ｐＭＳ－３１ｂ,在大肠杆菌ｐｏｐ２１３６中高效表达出人γ心钠素融合蛋白,表达产物占菌体总蛋白的３０～４０％。双向免疫扩散法测定证明表达产物与人α心钠素（α－ｈＡＮＰ）抗血清呈阳性反应,纯化的表达产物经复性处理后能引起大鼠胸主动脉条舒张．     

人脑利钠肽的Fmoc固相合成

探讨生物活性肽人脑利钠肽(BNP)的固相合成工艺,并为工业化合成提供理论依据.本文以二氯三甲基树脂(以下简称为二氯树脂)为载体,采用9-芴甲氧羰基(Fmoc)保护的氨基酸,以1-氧-3-双二甲胺羰基苯骈三氮唑四氟化硼盐(TBTU)/1-羟基苯并三氮唑(HOBT)/二异丙基乙胺(DIEA)缩合,以碘作为环化试剂,用切割试剂将BNP粗品从树脂上切割下来.通过MALDI-MS质谱仪检测,所合成环肽的分子量与理论分子量一致,使用RP-HPLC液相色谱仪对合成的环肽进行纯化,得到的BNP纯度达到97%以上.本合成工艺具有快捷、简便、高效的特点,适合于大批量的生产目的肽.     

Structural Substrate Conditions Required for Neutral Endopeptidase-Mediated Natriuretic Peptide Degradation

Natriuretic peptides are cyclic vasoactive peptide hormones with great diagnostic and therapeutic relevance. The main catabolic pathway postulated for natriuretic peptides is the degradation by neutral endopeptidase (NEP). However, B-type natriuretic peptide has been found to be resistant to NEP. Here, we compared the degradation of various mature, truncated, and recombinant natriuretic peptides by NEP. The degradation was clearly dependent on the length of the N- or C-terminus as well as on distinct sequence differences within the essential loop structure of the natriuretic peptides. Based on these findings, we developed a model for the interaction of NEP and natriuretic peptides that enables new insights into the mode of action and prediction of substrates of NEP, a peptidase that plays a key role in crucial (patho-) physiological processes.     

Natriuretic Peptide Clearance Receptor Alleles and Susceptibility to Abdominal Adiposity

Objective: To test the association of the C(?55)A polymorphism of the natriuretic peptide clearance receptor (NPRC) with blood pressure (BP), overweight/obesity, and body fat distribution in a large male adult population. Research Methods and Procedures: The study population was from a cross‐sectional and follow‐up study of 787 untreated male participants in the 1994 to 1995 follow‐up examination of the Olivetti Heart Study in Naples (356 of whom were examined previously in 1975). BP and anthropometric measures were taken, and biochemical assays were performed. The NPRC gene polymorphism C(?55)A was evaluated by polymerase chain reaction and HgaI digestion. Results: In the whole study population, there was no difference in BP, BMI, and biochemical tests among genotypes. Considering an A(?55) recessive model of inheritance, the AA subjects had lower BMI and waist circumference and lower prevalence of overweight, obesity, and abdominal adiposity as compared with the CC+CA subjects. On reviewing the characteristics of the subgroup previously examined in 1975, the AA subjects had already lower BMI, and their 20‐year rate of overweight and obesity was lower than the CC+CA subjects; no difference was observed in the rate of hypertension. Discussion: Male subjects carrying the A(?55)A NPRC genotype had a significantly lower prevalence of overweight, obesity, and abdominal adiposity. They also had a lower 20‐year rate of overweight compared with CC+CA individuals. These findings from a large unselected and untreated male population are in keeping with the recent evidence of a powerful lipolytic and lipomobilizing activity of natriuretic peptides.     

B型钠尿肽与心力衰竭研究现状

B型钠尿肽作为诊断充血性心力衰竭的一种生化指标,受到临床工作者的高度关注.本文详细综述了B型钠尿肽的生物特性、体内代谢、生理功能、实验室检测和在心力衰竭诊断、鉴别诊断、治疗、预后评价的应用.以及B型钠尿肽在实验室检测和临床应用中存在的问题.     

重组人脑利钠肽的临床应用进展

近年来,随着心力衰竭、肺动脉高压的病理生理及分子机制的深入研究,使上述疾病在临床药物治疗方面有了很大的进步,其中人脑利钠肽(BNP)作为体内唯一天然的肾素-血管紧张素-醛固酮拮抗剂在诊断及治疗心力衰竭等方面均引起了广泛关注,但由于其在心衰状态下降解快且生物活性明显减弱而限制了临床应用。因此,在心力衰竭治疗上补充外源性BNP成为了又一研究热点。重组人脑利钠肽(rhBNP)是一种人工合成的内源性激素,具有扩张血管、排钠利尿、降低心脏前后负荷、抑制肾素-血管紧张素-醛固酮系统和交感神经系统等作用,能够有效的改善充血性心力衰竭患者的血流动力学障碍。新近研究表明,rhBNP在治疗心血管疾病方面疗效显著,本文将就其在临床中的应用予以综述。     

Enhanced Activation of Atrial Natriuretic Peptide (ANP) and Natriuretic Peptide Receptor-A (NPRA) in Chronic Cigarette Smoke-Induced Lung Inflammation in Experimental Rats

International Journal of Peptide Research and Therapeutics - We sought to determine the circulatory level of atrial natriuretic peptide (ANP) and its receptor (Natriuretic peptide receptor-A) on...     

A Familial Mutation Renders Atrial Natriuretic Peptide Resistant to Proteolytic Degradation

A heterozygous frameshift mutation causing a 12-amino acid extension to the C terminus of atrial natriuretic peptide (ANP) was recently genetically linked to patients with familial atrial fibrillation (Hodgson-Zingman, D. M., Karst, M. L., Zingman, L. V., Heublein, D. M., Darbar, D., Herron, K. J., Ballew, J. D., de Andrade, M., Burnett, J. C., Jr., and Olson, T. M. (2008) N. Engl. J. Med. 359, 158–165). The frameshift product (fsANP), but not wild-type ANP (wtANP), was elevated in the serum of affected patients, but the molecular basis for the elevated peptide concentrations was not determined. Here, we measured the ability of fsANP to interact with natriuretic peptide receptors and to be proteolytically degraded. fsANP and wtANP bound and activated human NPR-A and NPR-C similarly, whereas fsANP had a slightly increased efficacy for human NPR-B. Proteolytic susceptibility was addressed with novel bioassays that measure the time required for kidney membranes or purified neutral endopeptidase to abolish ANP-dependent activation of NPR-A. The half-life of fsANP was markedly greater than that of wtANP in both assays. Additional membrane proteolysis studies indicated that wtANP and fsANP are preferentially degraded by neutral endopeptidase and serine peptidases, respectively. These data indicate that the familial ANP mutation associated with atrial fibrillation has only minor effects on natriuretic peptide receptor interactions but markedly modifies peptide proteolysis.Natriuretic peptides are pleiotropic factors that regulate blood pressure, cardiac hypertrophy, and long bone growth (1). Humans express three family members, atrial natriuretic peptide (ANP),³ B-type natriuretic peptide, and C-type natriuretic peptide (CNP). Each peptide is the product of a separate gene and contains a highly conserved 17-amino acid disulfide-linked ring structure that is required for biological activity. Atrial stretch causes ANP to be released from stored granules as a result of cardiovascular stresses like congestive heart failure. Once released into the circulation, ANP binds receptors in multiple tissues to reduce the load on the heart by stimulating natriuresis, diuresis, extravasation, vasorelaxation, and inhibiting the renin-angiotensin-aldosterone system (1).Natriuretic peptides exert their effects by binding one or more of three natriuretic peptide receptors. Natriuretic peptide receptor A (NPR-A) is the endogenous receptor for ANP and BNP (2, 3). NPR-A is a transmembrane guanylyl cyclase that, upon ligand binding, synthesizes the second messenger cGMP that mediates the renal and vascular effects of ANP and BNP (4, 5). Meanwhile, natriuretic peptide receptor B (NPR-B) is the receptor for CNP (6). NPR-B is highly homologous to NPR-A and also possesses guanylyl cyclase activity. The primary ligand for NPR-B is CNP, but this receptor can also be activated by very high concentrations of ANP or BNP (6). CNP-dependent activation of NPR-B stimulates long bone growth and may also inhibit cardiac hypertrophy (7). The third natriuretic peptide receptor is natriuretic peptide receptor C (NPR-C). Unlike NPR-A and NPR-B, NPR-C does not contain a guanylyl cyclase domain (8). Instead, the primary function of NPR-C is to control local natriuretic peptide concentrations through receptor-mediated internalization and degradation. Thus, it is typically referred to as the clearance receptor (9). In addition to its role in clearing natriuretic peptides from the circulation, NPR-C has also been shown to signal in a G protein-dependent manner (10). Finally, the other mechanism for natriuretic peptide removal is proteolytic degradation. Neutral endopeptidase (EC 3.4.24.11), which is also referred to as neprilysin or NEP, has been suggested to be the primary ANP-degrading enzyme in tissues associated with ANP clearance (11–13). Furthermore, inhibitors of NEP have been shown to increase circulating concentrations of ANP in rats (14).In a recent New England Journal of Medicine article, Hodgson-Zingman et al. (15) investigated the genetic basis for early onset atrial fibrillation in a family with white European ancestry. Using linkage analysis they found that all affected family members contained a single allele with a frameshift mutation in the coding portion of the ANP gene. The mutation causes a two-base pair deletion in exon 3 that eliminates the original stop codon and causes 12 new amino acids to be appended to the C terminus of the mature peptide. Thus, the peptide resulting from the frameshift mutation (fsANP) consists of 40 amino acids, whereas the wild-type peptide (wtANP) consists of 28 amino acids (Fig. 1). Importantly, the plasma levels of fsANP were shown to be 5–10-fold higher than the plasma concentrations of wtANP in affected individuals.Open in a separate windowFIGURE 1.Cartoon schematic of the primary amino acid structure of human atrial natriuretic peptide (wtANP) and the primary amino acid structure of the ANP frameshift mutation (fsANP). The 12-amino acid extension of fsANP is shaded in light gray. The dark gray shading indicates residues conserved in all natriuretic peptides. The black bars indicate disulfide bonds.Although Hodgson-Zingman et al. elegantly identified the ANP mutation associated with patients with early onset atrial fibrillation, they did not determine how this mutation affects the ability of ANP to interact with its known biological partners or why this mutation leads to elevated peptide concentrations. Theoretically, modulated binding to NPR-A, NPR-B, or NPR-C or altered proteolytic processing of ANP could lead to the observed disease. In this report, we identified subtle differences in the ability of fsANP and wtANP to interact with natriuretic peptide receptors but major differences in the proteolytic degradation of these peptides.     

慢性心力衰竭患者N端脑钠肽前体、脑钠肽与患者心脏超声参数及炎症因子的关系

目的:探讨慢性心力衰竭(CHF)患者N端脑钠肽前体(NT-proBNP)、脑钠肽(BNP)与患者心脏超声参数及炎症因子的关系。方法:选取2015年3月～2018年2月期间我院收治的CHF患者135例为研究组。根据纽约心脏病学协会(NYHA)心功能分级将研究组分为Ⅰ级组26例,Ⅱ级组34例,Ⅲ级组42例,IV级组33例。另选取同期于我院体检的健康志愿者30例为对照组。检测并比较对照组与研究组、不同NYHA心功能分级的血清标志物、炎症因子、心功能超声参数,采用Pearson相关性分析NT-proBNP、BNP与炎症因子、心功能超声参数的相关性。结果:研究组血清BNP、NT-proBNP、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)、超敏C反应蛋白(hs-CRP)水平及左心室舒张末期内径(LVEDD)、左心室收缩末期内径(LVESD)均显著高于对照组,左心室射血分数(LVEF)则低于对照组(P0.05)。BNP、NT-proBNP、IL-6、TNF-α、hs-CRP水平及LVEDD、LVESD随心功能分级的升高而升高,LVEF随心功能分级的升高而降低(P0.05)。经Pearson相关性分析显示,CHF患者BNP、NT-proBNP与IL-6、TNF-α、hs-CRP、LVEDD、LVESD呈正相关,而与LVEF呈负相关(P0.05)。结论:血清NT-proBNP、BNP与CHF患者的心脏超声参数及炎症因子密切相关,可考虑将其作为临床诊断CHF的生物学指标。     

B型利钠肽在心衰应用的研究进展

利钠肽（natriuretic peptides，NPs）的发现已有30年历史，其中B型利钠肽（B-type natriuretic peptide，BNP）及氨基末端脑钠肽前体（ N-terminal pro-B type natriuretic peptide，NT-proBNP）的临床应用对心血管疾病诊治具有里程碑意义，特别是对心衰的综合价值最高。BNP具备强大的心血管保护效应，但心衰后BNP水平大幅升高，却没展现出相应的活性，被称为“利钠肽悖论”。近几年，随着质谱、核磁共振技术的运用，逐渐从代谢途径和检测技术上解开了“利钠肽悖论”这一谜题：外周循环中存在多种不同生物活性的BNP亚型且心衰后的BNP代谢与生理状态下不同。所以，纵然检测到心衰后BNP大幅升高，但本质上是由于传统检测技术的瓶颈，使各类BNP亚型与检测试剂交叉反应，活性成分被高估而造成假阳性。因此，要加强对BNP在病理生理等不同情况下的认识，还要借助生物化学手段建立具有敏感性和特异性的检测方法来识别BNP_1-32、BNP_1-30、BNP_3-32及B型利钠肽原（pro-B-type natriuretic peptide，proBNP）等特殊形式。从而有助于探索心衰更深层次的病理生理机制，还可协助临床对心衰的诊断及预后做出更准确的判断。     

恒河猴心房利钠多肽的分离、纯化及活性研究

<正> 1983年以来,人们相继从人和大鼠等动物的心房中分离到心房利钠多肽(ANP)。但对非人灵长类动物——恒河猴的ANP了解甚少。由于ANP对机体循环系统的调节起着重要作用,其强大的降压、利钠、利尿的生理功能,在心血管疾病的防治方面具有重要意义。我们曾报导过恒河猴心房肌细胞中存在着大量的ANP样物质,随后对此物质进行了分离、纯化及活性检测,现简报如下。     

B-Type Natriuretic Peptide and Prognosis of End-Stage Renal Disease: A Meta-Analysis

Background

The prognostic importance of B-type natriuretic peptide (BNP) or N-terminal pro BNP (NT-proBNP) in patients with end-stage renal disease (ESRD) remains controversial.

Methodology/Principal Findings

We conducted an unrestricted search from the MEDLINE and EMBASE in all languages that were published between 1966 and Augest2013. Twenty-seven long-term prospective studies met our inclusion criterias. From the pooled analysis, elevated BNP/NT-proBNP was significantly associated with increased all cause mortality [odds ratio (OR), 3.85; 95% CI, 3.11 to 4.75], cardiovascular mortality (OR, 4.05; 95% CI, 2.53 to 6.84), and cardiovascular events (OR, 7.02; 95% CI, 2.21 to 22.33). The funnel plot showed no evidence of publication bias. The corresponding pooled positive and negative likelihood ratio for prediction of all cause mortality were 1.86 (95% CI, 1.66 to 2.08) and 0.48 (95% CI, 0.42 to 0.55), respectively.

Conclusions/Significance

BNP/NT-proBNP is a promising prognostic tool to risk-stratify the patients with ESRD. Further investigations are warranted to elucidate the specific pathogenic mechanisms and the impact of other potential prognostic factors.     

Atrial Natriuretic Peptide Regulates Ca2+ Channel in Early Developmental Cardiomyocytes

Background

Cardiomyocytes derived from murine embryonic stem (ES) cells possess various membrane currents and signaling cascades link to that of embryonic hearts. The role of atrial natriuretic peptide (ANP) in regulation of membrane potentials and Ca²⁺ currents has not been investigated in developmental cardiomyocytes.

Methodology/Principal Findings

We investigated the role of ANP in regulating L-type Ca²⁺ channel current (I_CaL) in different developmental stages of cardiomyocytes derived from ES cells. ANP decreased the frequency of action potentials (APs) in early developmental stage (EDS) cardiomyocytes, embryonic bodies (EB) as well as whole embryo hearts. ANP exerted an inhibitory effect on basal I_CaL in about 70% EDS cardiomyocytes tested but only in about 30% late developmental stage (LDS) cells. However, after stimulation of I_CaL by isoproterenol (ISO) in LDS cells, ANP inhibited the response in about 70% cells. The depression of I_CaL induced by ANP was not affected by either Nω, Nitro-L-Arginine methyl ester (L-NAME), a nitric oxide synthetase (NOS) inhibitor, or KT5823, a cGMP-dependent protein kinase (PKG) selective inhibitor, in either EDS and LDS cells; whereas depression of I_CaL by ANP was entirely abolished by erythro-9-(2-Hydroxy-3-nonyl) adenine (EHNA), a selective inhibitor of type 2 phosphodiesterase(PDE2) in most cells tested.

Conclusion/Significances

Taken together, these results indicate that ANP induced depression of action potentials and I_CaL is due to activation of particulate guanylyl cyclase (GC), cGMP production and cGMP-activation of PDE2 mediated depression of adenosine 3′, 5′–cyclic monophophate (cAMP)–cAMP-dependent protein kinase (PKA) in early cardiomyogenesis.