Bio-Molecular Markers for Cardiovascular Disease:Significance of Natriuretic Peptides and Adrenomedullin

Takeshi Horio,Yuhei Kawano 대한심장학회 2008 Korean Circulation Journal Vol.38 No.10

There are many established and proposed bio-molecular markers for cardiovascular disease, including vasoactive substances, substances related to inflammation and oxidative stress, and substances involved in tissue structure and remodeling. Among these substances, we focused on natriuretic peptides and adrenomedullin (AM) as clinically useful bio-molecular markers in this review. Three natriuretic peptides-atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP)-play various important roles in the cardiovascular system. ANP and BNP are released from the heart and exist primarily as circulating hormones. They participate in the regulation of blood pressure and fluid levels. Plasma levels of ANP and BNP are increased in various pathological conditions such as heart failure, myocardial infarction, and hypertension with cardiac hypertrophy. BNP is now essential as a biochemical marker in managing patients with cardiovascular disease. CNP is mainly produced in vascular endothelium. It contributes to smooth muscle relaxation and growth inhibition as a local hormone, but it is also synthesized in cardiac fibroblasts and inhibits fibroblast proliferation and myocyte growth. However, the significance of plasma CNP levels remains to be elucidated. AM is widely distributed in various organs and tissues, including the cardiovascular system. Not only it is a potent vasodilator peptide, but it also has protective effects against vascular and cardiac cell injury and excessive growth. Plasma AM levels are increased in several cardiovascular diseases, including hypertension, heart failure, myocardial infarction, and atherosclerotic disease, and AM appears to be a predictive and prognostic marker in the setting of cardiovascular disease. There are many established and proposed bio-molecular markers for cardiovascular disease, including vasoactive substances, substances related to inflammation and oxidative stress, and substances involved in tissue structure and remodeling. Among these substances, we focused on natriuretic peptides and adrenomedullin (AM) as clinically useful bio-molecular markers in this review. Three natriuretic peptides-atrial natriuretic peptide (ANP), brain natriuretic peptide (BNP), and C-type natriuretic peptide (CNP)-play various important roles in the cardiovascular system. ANP and BNP are released from the heart and exist primarily as circulating hormones. They participate in the regulation of blood pressure and fluid levels. Plasma levels of ANP and BNP are increased in various pathological conditions such as heart failure, myocardial infarction, and hypertension with cardiac hypertrophy. BNP is now essential as a biochemical marker in managing patients with cardiovascular disease. CNP is mainly produced in vascular endothelium. It contributes to smooth muscle relaxation and growth inhibition as a local hormone, but it is also synthesized in cardiac fibroblasts and inhibits fibroblast proliferation and myocyte growth. However, the significance of plasma CNP levels remains to be elucidated. AM is widely distributed in various organs and tissues, including the cardiovascular system. Not only it is a potent vasodilator peptide, but it also has protective effects against vascular and cardiac cell injury and excessive growth. Plasma AM levels are increased in several cardiovascular diseases, including hypertension, heart failure, myocardial infarction, and atherosclerotic disease, and AM appears to be a predictive and prognostic marker in the setting of cardiovascular disease.

자라 신장기능에 미치는 Atrial Natriuretic Peptide의 효과

조경우,김선희,고규영,설경환,Cho, Kyung-Woo,Kim, Suhn-Hee,Koh, Gou-Young,Seul, Kyung-Hwan 대한생리학회 1987 대한생리학회지 Vol.21 No.1

Effects of synthetic atrial natriuretic peptide and furosemide on the cardiovascular and renal functions were examined in the freshwater turtle, Amyda japonica. Both atria and ventricle of turtle contained an immunoreactive atrial natriuretic peptide. Synthetic rat atrial natriuretic peptide (atriopeptin III) and turtle atrial extract caused a decrease in mean arterial blood pressure and the vasodepressor effect was dose-dependent. In hydrated turtles received either atriopeptin III or turtle atrial extract, no significant change in renal function was observed until 100 min except a slight natriuresis at 60 or 100 min after injection of 30 ug/kg atriopeptin III or atrial extract, respectively. However, furosemide, 2 mg/kg, caused marked diuresis, natriuresis and kaliuresis. In non-hydrated turtles, no significant change in renal function was observed until 6 hrs following injection of 30 ug/kg atriopeptin III. Plasma aldosterone decreased at 2 hr and increased at 24 hr after injection of atriopeptin III although plasma renin concentration did not change. But, furosemide caused persistent diuresis, natriuresis and kaliuresis. Additionally, plasma aldosterone and renin concentrations were significantly increased at 24 hrs after injection of furosemide. In conclusion, we suggest that the freshwater turtle may have an atrial natriuretic peptide in heart and vascular receptors for atrial natriuretic peptide, and that atrial natriuretic peptide is more important in the regulation of blood pressure rather than that of renal function in freshwater turtles. We also suggest that an increased plasma renin concentration caused by furosemide may not be due to the sodium concentration delivered to macula densa, but due to the dehydration caused by persistent diuresis and natriuresis.

생쥐 턱밑샘의 발생, 분화 및 노화에 따른 C-type natriuretic peptide와 natriuretic peptide 수용기의 발현양상

복세미(Se-Mi Bok),김택헌(Tak-Heun Kim),박기랑(Kee-Rang Park),조의식(Eui-Sic Cho) 대한해부학회 2005 Anatomy & Cell Biology Vol.38 No.4

C-type natriuretic peptide (CNP)는 natriuretic peptide family의 하나로서 주로 중추신경계통과 혈관내피세포에서 국소적으로 합성되는 것으로 알려져 있으나 최근 다른 말초기관에서도 합성되어 세포의 성장과 분화 조절에 관여하는 것으로 알려지고 있다. 본 연구자들은 CNP가 정상 성체 턱밑샘의 사이관과 곱슬과립세관세포에서 국소적으로 합성되어 분포하며 생물학적 활성을 보임을 보고한 바 있으나, 발생 및 분화, 그리고 노화에 따르는 발현 및 활성에 대해서는 알려진 바 없다. 따라서 본 연구는 턱밑샘의 발생 및 분화과정에서 CNP와 그 수용기의 발현양상과 노화과정에서 그 수용기의 생물학적 활성변화를 알아보고자, 임신14, 16, 18일된 C57BL/6N 생쥐 배자와 출생 후 1일, 2주, 그리고 1, 2, 12, 24개월된 성체의 턱밑샘을 대상으로 RT-PCR, in situ hybridization, 면역조직화학법 그리고 cGMP assay를 시행하였다. CNP는 임신14일에 턱밑샘의 원기인 외배엽성상피싹 종말부위에서 강하게 발현되었으며 출생 전까지 높게 유지되었으나 출생 후 현저히 감소되었다. CNP는 성체 턱밑샘의 사이관과 곱슬과립세관에 분포하고 있었으며 이들 관계통에서 NPRC는 특이적으로 발현되었으나 NPRB는 발현되지 않았다. RT-PCR결과 턱밑샘에서 CNP전사체는 임신16일부터 출생 후 2개월까지 점차 감소하였으나 ANP전사체는 증가하였다. CNP에 특이적인 수용기인 NPRB와 NPRC는 CNP와 동일한 발현양상이었으나 NPRA는 약하게 발현되었다. 또한 CNP는 치아싹, 혀, 앞위턱뼈, 뼈형성 부위와 같은 두개안면 조직에서도 발현되었으며, 이들 부위에서도 NPRC는 발현되었으나 NPRB는 발현되지 않았다. 출생 후 2개월된 턱밑샘의 관세포는 CNP에 의해 현저히 많은 cGMP를 생성하였으나, 샘꽈리는 CNP보다는 ANP와 BNP에 의해 많은 cGMP를 생성하였다. 노화에 따라 턱밑샘의 cGMP 활성능력은 전반적으로 감소되었으며 출생 후 1개월된 턱밑샘에서는 CNP, 출생 후 2개월 이후로는 ANP에 의한 cGMP 활성이 우세하였다. 이상의 결과로 보아 CNP는 턱밑샘에서 NPRC를 매개로 하는 발생 및 분화유도 기능을 주로 수행하며, 성체이후에는 NPRB를 매개로 하는 관계통 유지에 관여하고 노화에 따라서 활성이 점차 감소하는 것으로 추정된다. C-type natriuretic peptide (CNP), a member of natriuretic peptide family, is mainly synthesized in the endothelium and central nervous system. But CNP is also involved in the growth and differentiation of other peripheral organs. Although we have reported the local synthesis and localization of CNP in the adult submandibular glands (SMG), it is not known that the expression and biological activity of CNP following the morphogenesis, differentiation and aging. This study aimed to examine the expression of CNP and its receptors in the developing and differentiating stages of mouse SMG, and the changes of biological activity of its receptors with aging. The SMG, obtained from 14, 16, 18 days-old embryos (E) and 1 day, 2 weeks, 1, 2, 12, and 24 month-old C57BL/6N mouse, were processed for RT-PCR, in situ hybridization, immunohistochemistry and cGMP assay. CNP was strongly expressed in the epithelial clusters of primitive SMG, which was maintained before birth but was markedly decreased after birth. CNP was localized in the intercalated duct and granular convoluted tubules of adult SMG, where NPRC was specifically expressed but NPRB was not. CNP mRNA was gradually decreased from E16 to 2 M but ANP mRNA was opposed. NPRB and NPRC were the same pattern of the expression of CNP but NPRA was weakly expressed. In addition, CNP mRNA was also expressed in the craniofacial tissues such as tooth germs, tongue, premaxilla and bone forming area in which NPRC was specifically expressed but NPRB was not. In the SMG of 2 M, the membrane of duct cells markedly produced cGMP by CNP whereas acini produced cGMP by ANP and BNP rather than CNP. The biological activity of cGMP production of SMG gradually decreased with age. cGMP production was dominant by CNP in SMG of 1M but was by ANP after 2M. These results shows that CNP may play roles both in the morphogenesis and differentiation via NPRC and in the maintenance of duct system via NPRB in the mouse SMG and that the biological activity of its receptors may decreased with aging.

흰쥐 심장조직에서 식염부하에 의한 Natriuretic Peptides의 변화에 대한 면역조직화학적 연구

김두일,김동수 인제대학교 1998 仁濟醫學 Vol.19 No.2

Atrial natriuretic peptide(ANP)는 나트륨 이뇨, 이뇨 및 혈관 확장 등의 작용을 가지고 있는 심장 호르몬으로서 체액, 전해질 및 혈압의 항상성을 유지하며, brain natriuretic peptide(BNP) 역시 신경펩티드로서 ANP와 유사한 작용을 가지는 호르몬으로 알려져 있다. 본 연구에서 저자들은 흰쥐에서 식염부하를 시도하여 심장조직에서 면역조직화학염색법으로 ANP와 BNP의 면역반응성의 분포를 조사하여 그것의 활성도 및 반응의 지속 정도를 알아보고, 또한 혈장 삼투질 농도 및 Na+ 농도 변화를 관찰하였다. Objectives: Atrial natriuretic peptide(ANP) is a cardiac hormone with diuretic, natriuretic, and vasorelaxant activities, and it is implicated in the control of blood pressure, body fluid, and cardiovascular homeostasis. Brain natriuretic peptide(BNP) forms a peptide family with ANP and may be involved in the regulation of blood pressure and fluid volume, as is ANP. It has been reported that natriuretic responses are altered by changes in extracellular fluid(ECF) volume. Methods and Materials: To assess the compensatory mechanism of ANP and BNP in response to ECF volume alterations, we examined the distribution of immunohistochemically identified cells for ANP and BNP in the heart tissues of salt-loaded rats using immunohistochemical method and also investigated the process, activity, maintenance of reaction during salt intake. In addition, plasma osmolality, plasma sodium concentration were measured in association with changes in salt intake. Results: 1. Plasma osmolality increased wish advancing salt intake, but plasma sodium concentration was greater than the comparable values in the control group at the third day of salt intake. 2. The distribution of ANP-immunoreactivity in the heart was wider in the right atrial tissue. The ANP-immunoreactivity was more intense in right aerial tissue, interventricular septum and coronary sinus at the third day of salt intake and became weaker from the seventh day of salt intake. The BNP-immunoreactivity in the heart tissue was observed a group of many immunopositive cells in right atrial tissue, left ventricular tissue, interventricular septum at the fifth day of salt intake. Thereafter it was decreased gradually and seemed to be exhausted from the tenth day of salt intake. Conclusion: ANP and BNP are factors that maintain the homeostatic balance of body fluid during salt intake. However further investigation on these plasma levels, receptor binding, and biological activity is required to exactly understand its participation and importance in the natriuretic peptides hormonal system.

Localization of Dendroaspis natriuretic peptide (DNP) in the rat salivary glands

Kim, Jae-Gon,Lee, Young-Soo,Baik, Byeong-Ju,Park, Byung-Keon 대한소아치과학회 2001 大韓小兒齒科學會誌 Vol.28 No.3

Dendroaspis natriuretic peptide(DNP)는 Dendroaspis angusticeps의 뱀독으로부터 추출된 natriuretic peptide의 새로운 일원으로서, 사람의 혈장과 심방 근육에 존재하고 실험동물에서 혈관확장과 이뇨작용을 나타내는 것으로 알려져있다. 그러나 아직까지 심장 이 외의 다른 기관내에 DNP가 존재하는 지 또한 이들이 현재까지 알려진 기능 이 외의 또다른 생리적 기능을나타내는지의 여부는 아직 확실하게 밝혀져 있지 않다. 타액전 내에서 DNP의 기능을 규명하기 위한 전단계로, 먼저 횐쥐의 타액선내에 DNP가 존재하는지를 면역조직화학법을 이용하여 알아보았고 또한 DNP의 수용체가 타액선 또는 구강점막내에 존재하는 지의 여부를 생체의 방사선자기 법을 이용하여 알아보았다. DNP의 면역반응은 악하선과 설하선 및 이하선에서 주로 도관부위에서 관찰되었고, 선포부위에서는 관찰되지 않았다. atrial natriuretic peptide(ANP)의 면역반응 역시 악하선과 설하선 및 이하선에서 DNP면역반응과 유사하게 주로 도관부 위에서 관찰되었다 DNP에 특이한 수용체가 타액선에는 저밀도로 존재한 반면 경구개와 혀 점막상피부위에서는 고밀도로 존재하였다. 경구개와 혀 점막상피부위의 DNP수용체는 주로 상피의 기저층과 기저층에 인접한 유극세포층에 국한되어 존재하는 것을 미세방사선자기 법을 통하여 알 수 있었다. 이러한 결과로 볼 때 DNP가 타액선과 구강점 막 부위에서 어떠한 생리적 기능을 나타낼 것으로 여겨지는데 , 특히 구강점 막부위에서는 점막상피세포의 성장과 성숙과정에 관여하리라 추정된다. Dendroaspis natriuretic peptide (DNP), a fourth member of the natriuretic peptide isolated from the venom of the Dendroaspis angusticeps snake, has been reported to be present in human plasma and atrial myocardium and caused vasorelaxation and diuresis in experimental animals. However. it is uncertain whether they are present in peripheral organs other than the heart and its further physiological roles also remains to be clarified. To assess the possible physiological role of DNP in the salivary glands, I investigated the localization of DNP peptide in the rat salivary glands by immunohistochemistry and the binding sites for radiolabelled DNP in the rat salivary glands and oral mucosa using in vitro autoradiography. DNP immunoreactivity was widely distributed in the submandibular. sublingual and parotid glands, particularly in the ducts such as the intercalated and striated ducts, where atrial natriuretic peptide (ANP) was colocalized in consecutive sections, but not in acini. High density 125I-DNP binding sites were localized in the epithelia of the tongue and hard palate, while low density binding sites fur 125I-DNP were also distributed in the submandibular, sublingual, and parotid glands. In the hard palate and tongue, the precise location of this binding was revealed on the basal and parabasal cells of the epithelia by emulsion microautoradiography. These results suggest that DNP may not only have a role in the salivary glands but also play a role in the regulation of growth in the oral epithelium, particularly in the hard palate and tongue.

Spontaneously Hypertensive Rat에 있어서 Atrial Natriuretic Peptide의 신장기능과 몇가지 호르몬 분비에 미치는 영향

김산호(Kim, San-Ho),김선희(Kim, Suhn-Hee),설경환(Seul, Kyung-Hwan),조경우(Cho, Kyung-Woo) 대한생리학회 1988 대한생리학회지 Vol.22 No.1

The present study was undertaken to clarify the involvement of atrial natriuretic peptide in the development of hypertension in spontaneously hypertensive rats. Plasma concentration of immunoreactive atrial natriuretic peptide was higher in spontaneously hypertensive rats than in normotensive Sprague-Dawley and Wistar rats. Plasma renin concentration was lower in SHR than in normotensive rats, as observed in earlier experiments. Hydration-induced increase in urine flow and urinary excretions of sodium and potassium were smaller in SHR than in normotensive control rats. Intraarterial infusion of atrial natriuretic peptide resulted in increases in urine flow, urinary excretions of sodium and potassium in both hypertensive and normotensive rats. Renal response to atrial natriuretic peptide was markedly suppressed in SHR. Plasma renin and aldosterone concentration were suppressed by atrial natriuretic peptide in both SHR and normotensive rats. The responses were not significantly different in both groups. These results suggest that the renal responsiveness to atrial natriuretic peptide may be suppressed in SHR by some mechanisms still remaining obscure.

지구성 운동이 심혈관 질환 예측인자인 ANP & BNP에 미치는 영향

김선호(Kim Sun-Ho),윤영복(Yoon Young-Bok) 한국체육과학회 2007 한국체육과학회지 Vol.16 No.3

The aim of this study was to investigate the influence of endurance training in atrial natriuretic peptide & brain natriuretic peptide. For this study, 16 healthy young men were selected. Subjects participated in 12 weeks of running to four times per week for 30-40minutes. Statistical techniques for data analysis were a descriptive statistics and repeated ANOVA. The results of this study were as following; 1. Atrial natriuretic peptide was shown a case of decreasing after 6 weeks, but after 12 weeks have decreased significantly. Especially, ANP was significantly lower after 12 weeks than after both 6 weeks and 0 weeks at all the blood sampling time. 2. Brain natriuretic peptide was shown a case of decreasing after 6 weeks at all the blood sampling time, but there was no significant difference. Also, BNP was significantly lower after 12 weeks than after both 6 weeks and 0 weeks at post-maximal exercise These data suggest that endurance exercise significantly decreased ANP & BNP levels in healthy men, and the decreases could be partially attributed to cardiovascular functions.

Nifedipine이 Atrial Natriuretic Peptide의 혈압내림효과에 미치는 영향

이종은(Lee, Jong-Eun),최기철(Choi, Ki-Chul) 대한생리학회 1990 대한생리학회지 Vol.24 No.1

Pentobarbital 마취한 정상혈압 및 신성 고혈압 흰쥐에서 calcium channel 봉쇄 약물 nifedipine과 atrial natriuretic peptide(ANP)의 상호작용을 조사하였다. 정상혈압 흰쥐에서 nifedipine(1.0 μg/kg/min) 또는 ANP(60 ng/kg/min)의 주입은 각각 유의하게 혈압을 내렸으며 두 약물의 동시 주입시에 개별적으로 주입하였을 때보다 그 혈압내림의 정도가 더욱 컸다. Nifedipine은 단독 주입하였을 때에 신기능에 유의한 영향을 미치지 않았으나 ANP와 동시에 주입하였을 때에는 ANP의 요량 및 Na 배설 증가 효과를 항진시켰다. 한편 고혈압 흰쥐에서도 ANP의 혈압내림효과와 신장효과는 nifedipine과 함께 주입하였을 때에 더 컸다. 적출 흉부대동맥 표본을 phenylephrine으로 미리 수축시킨 후 ANP를 첨가하면 용량의존 이완반응을 보였고 nifedipine 존재하에서 더 예민하였다. 이상의 실험결과는 calcium channel 봉쇄약물이 ANP의 혈압내림효과를 항진시킴을 보인 것이며 그 기전으로 요중 배설 증가 및 혈관이완효과 증가 등이 관여함을 시사하였다. The interaction between a calcium channel blocker nifedipine and atrial natriuretic peptide (ANP) was examined in normotensive and renal hypertensive rats. The infusion of either ANP or nifedipine produced a significant decrease in mean arterial pressure (MAP). The combined infusion of ANP with nifedipine resulted in a greater fall of MAP than did the infusion of each drug alone. ANP significantly increased urinary volume and excretion of sodium, while nifedipine was without effects. The diuretic/natriuretic effects of ANP were potentiated by the combined infusion with nifedipine. The vasodepressor and renal effects of ANP or nifedipine were qualitatively similar between the normotensive and hypertensive rats. Nifedipine caused an upward and leftward shift of the ANP dose-relaxation curve of the phenylephrine-precontracted thoracic aortic rings isolated from the normotensive rats , suggesting that the vasodilation sensitivity to ANP is increased in the presence of nifedipine. These results indicate that nifedipine enhances the vasodepressor effect of ANP, the likely mechanisms being attributable to a contraction of effective intravascular volume as a consequence of potentiated renal excretion and a greater peripheral vasodilation.

본태성 고혈압환자에 있어서 Atrial Natriuretic Peptide와 Brain Natriuretic peptide 농도에 관한연구

류혜영,김우식,황석재,강흥선,조정휘,김권상,송정상,배종화 대한순환기학회 2000 Korean Circulation Journal Vol.30 No.7

High Plasma Levels of the B-type Natriuretic Peptide in Patients Without Heart Failure: Is There Clinical Significance?

Long Hao Yu,김무현,박종성,차광수,박태호,김영대 대한심장학회 2010 Korean Circulation Journal Vol.40 No.3

We report a case of a 19-year-old female with an elevated plasma B-type natriuretic peptide (BNP) level, but without evidence of heart failure (HF). She presented with non-specific chest pain and a high level of the B-type natriuretic peptide, despite having unremarkable findings on physical examination, laboratory analysis, electrocardiogram,echocardiogram, chest X-ray, chest computed tomography, whole body scan, and coronary angiography. We attribute this finding to a genetic variation in the synthesis and cleavage of the natriuretic peptides.