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Endocrinology Vol. 139, No. 11 4576-4580
Copyright © 1998 by The Endocrine Society

ARTICLES

Production and Secretion of Adrenomedullin in Cultured Rat Cardiac Myocytes and Nonmyocytes: Stimulation by Interleukin-1ß and Tumor Necrosis Factor-{alpha}1

Takeshi Horio, Toshio Nishikimi, Fumiki Yoshihara, Noritoshi Nagaya, Hisayuki Matsuo, Shuichi Takishita and Kenji Kangawa

Division of Hypertension, Department of Medicine (T.H., S.T.), and Research Institute (T.N., F.Y., N.N., H.M., K.K.), National Cardiovascular Center, Suita, Osaka 565-8565, Japan

Address correspondence and requests for reprints to: Takeshi Horio, M.D., Division of Hypertension, Department of Medicine, National Cardiovascular Center, 5–7-1, Fujishirodai, Suita, Osaka 565-8565, Japan.


   Abstract
Top
 Abstract
Introduction
 Materials and Methods
 Results
 Discussion
 References
 
The present study investigated the secretion level and gene expression of adrenomedullin (AM), a novel vasorelaxant peptide, in cultured neonatal rat cardiac myocytes and nonmyocytes, and the effects of interleukin-1ß (IL-1ß) and tumor necrosis factor-{alpha} (TNF{alpha}) on its production and secretion in these cells. Under serum-free conditions, both myocytes and nonmyocytes secreted immunoreactive (ir-) AM into the culture medium in a time-dependent manner. The secretion rates of ir-AM from myocytes and nonmyocytes per 105 cells were almost equivalent. The expression of AM messenger RNA was also observed in cultured myocytes and nonmyocytes. The peptide secretion and messenger RNA level of AM in cardiac myocytes were increased after stimulation with IL-1ß. In nonmyocytes, IL-1ß and TNF{alpha} remarkably augmented both the release of ir-AM into the medium and AM gene expression after 24 and 48 h of incubation. These observations indicate that cardiac ventricular cells (i.e. myocytes and nonmyocytes) actively produce AM and also suggest that cytokines such as IL-1ß and TNF{alpha} regulate the gene expression and secretion of this peptide in the ventricles. On the basis of these results and the findings that IL-1ß and TNF{alpha} are involved in heart failure and cardiac hypertrophy, AM may play a role as an autocrine/paracrine modulator in some cardiac disorders.


   Introduction
Top
 Abstract
 Introduction
Materials and Methods
 Results
 Discussion
 References
 
ADRENOMEDULLIN (AM) is a potent vasodilator peptide that was originally isolated from human pheochromocytoma (1). Subsequent studies have revealed that AM is produced and secreted from vascular endothelial cells and smooth muscle cells (2, 3). Inversely, AM exerts some biological effects as well as a vasorelaxant action on vascular cells (4, 5, 6). These observations suggest that AM may act as a paracrine and/or autocrine hormone in the regulation of vascular homeostasis.

The presence of AM peptide and its gene expression have already been shown in the heart (2, 7, 8). A recent immunohistochemical study revealed that AM immunoreactivity is markedly increased in failing human ventricles (9). In addition, we demonstrated augmented levels of the AM gene and peptide in the hearts of rats with heart failure (10). These observations indicate that AM is produced from normal hearts, and that its production is accelerated in cardiac disorders such as heart failure. However, little is known about the production of AM in cultured cardiac cells, and it has not been elucidated which type of cells predominantly synthesizes AM in quiescent and stimulated conditions. Therefore, we conducted such a study, examining the peptide release and gene expression of AM in cultured ventricular myocytes and nonmyocytes (fibroblasts) of neonatal rat hearts. Interleukin-1ß (IL-1ß) and tumor necrosis factor-{alpha} (TNF{alpha}) have been reported to stimulate AM synthesis from cultured vascular smooth muscle cells (3, 11). As these cytokines may participate in cardiac dysfunction during heart failure and in the progression of cardiac hypertrophy (12, 13, 14), we also investigated the effects of IL-1ß and TNF{alpha} on the production and secretion of AM in cardiac myocytes and nonmyocytes.


   Materials and Methods
Top
 Abstract
 Introduction
 Materials and Methods
Results
 Discussion
 References
 
Cell culture
Primary cultures of neonatal ventricular myocytes and nonmyocytes were prepared according to the procedure described by Nakagawa et al. (15) with minor modifications. Apical halves of cardiac ventricles from 1- to 2-day-old Wistar rats were separated and minced in a chilled balanced salt solution (116 mM NaCl, 20 mM HEPES, 12.5 mM NaH2PO4, 5.6 mM glucose, 5.4 mM KCl, and 0.8 mM MgSO4, pH 7.35). Ventricular cardiocytes were dispersed in the balanced salt solution containing 0.06% collagenase type II (Worthington Biochemical Corp., Freehold, NJ) with agitation for 6 min at 37 C. The digestion steps were repeated five to seven times until the tissues were completely digested. The cells were combined, centrifuged, and resuspended in chilled FCS (Life Technologies, Grand Island, NY). To segregate myocytes from nonmyocytes, a discontinuous gradient of Percoll (Sigma Chemical Co., Inc., St. Louis, MO) consisting of 40.5% and 58.5% was prepared in the balanced salt solution, and ventricular cells were suspended in the layer of 58.5% Percoll. After centrifugation at 3000 rpm for 30 min, the upper layer consisted of a mixed population of nonmyocyte cell types, and the lower layer consisted almost exclusively of cardiac myocytes. Both myocytes and nonmyocytes were washed twice by centrifugation and resuspension to remove all traces of Percoll.

After myocytes were incubated twice on uncoated 10-cm culture dishes for 30 min to remove any remaining nonmyocyte, the nonattached viable cells (purified myocytes) were plated at a density of 4.0 x 105 cells/well onto gelatin-coated six-well tissue culture plates and cultured in DMEM (Life Technologies) supplemented with 10% FCS and antibiotics (50 U/ml penicillin and 50 µg/ml streptomycin, ICN Biomedicals, Inc., Aurora, OH) at 37 C for 48 h in humidified air with 5% CO2.

Nonmyocyte cells were resuspended in DMEM with 10% FCS and plated onto uncoated 10-cm culture dishes for 30 min. After the plating period, nonadherent cells and debris were washed away, and fresh medium was added. Cells were allowed to grow to confluence, trypsinized, and passaged 1:3. This procedure yielded cultures of cells that were almost exclusively fibroblasts by first passage as described by Villarreal et al. (16). Nonmyocytes at the second or third passage were plated onto six-well plates and allowed to grow to confluence (2.5 x 105 cells/well).

After incubation in DMEM with FCS, myocytes and nonmyocytes were maintained in serum-free DMEM for 12 h. After the preconditioning period, the cultured cells were replaced with fresh serum-free DMEM containing 1 mg/ml BSA (Seikagaku Corp., Tokyo, Japan). Then, IL-1ß (Genzyme Transgenics Corp., Cambridge, MA), TNF{alpha} (Sigma), angiotensin II (Peptide Institute, Inc., Osaka, Japan), endothelin-1 (Peptide Institute), phenylephrine (Research Biochemicals International, Inc., Natick, MA), or vehicle was added, and the plates were incubated for 6–48 h. After incubation, the medium was aspirated and stored at -80 C until RIA. After washing with PBS, the cells were submitted for RNA extraction.

Measurement of immunoreactive (ir-) AM and ir-atrial natriuretic peptide (ANP)
The culture medium (0.5 or 1 ml) was acidified with acetic acid, boiled for 5 min to inactivate intrinsic proteases, and lyophilized. RIA for rat AM or rat ANP was performed as previously reported (10, 17). The anti-AM antibody recognized the C-terminal region of rat AM and did not cross-react with rat ANP. The antibody against rat ANP did not also cross-react with rat AM. These assays were performed in duplicate.

Characterization of ir-AM in culture medium
The conditioned medium (40 ml) from myocytes or nonmyocytes was condensed with a Sep-Pak C18 cartridge (Waters Corp., Milford, MA) and separated by reverse phase HPLC on a µ-Bondasphere C18 column (300 Å, 3.9 x 150 mm; Waters Corp.). A linear gradient elution of acetonitrile for 10–60% in 0.1% trifluoroacetic acid was made at a flow rate of 1 ml/min, and each collected fraction (1 ml) was submitted for RIA for rat AM.

Northern blot analysis for rat AM and ANP messenger RNA (mRNA)
Total RNA was extracted from cultured cells by the acid guanidinium thiocyanate-phenol-chloroform method, according to the method previously reported (2). Total RNA (30 µg/lane) was denatured with formaldehyde and formamide, and electrophoresed on a 1% agarose gel containing formaldehyde. RNA in the gel was then transferred to a nylon membrane (Zeta-Probe blotting membrane, Bio-Rad Laboratories, Inc., Hercules, CA) and fixed by UV irradiation. Hybridization and washing of the membrane were carried out with complementary DNA (cDNA) probes for rat AM and rat ANP genes as described previously (10). Band intensity was estimated by a radioimage analyzer (BAS-5000, Fuji Photo Film Co., Ltd., Tokyo, Japan). For comparison of mRNA contents in each sample, the same membrane was rehybridized with a glyceraldehyde-3-phosphate dehydrogenase (GAPDH) cDNA probe.

Calculations and statistical analysis
The statistical significance of differences in the results was evaluated using an unpaired ANOVA, and P values were calculated by Scheffe’s method. P < 0.05 was accepted as statistically significant.


   Results
Top
 Abstract
 Introduction
 Materials and Methods
 Results
Discussion
 References
 
Basal secretion and characterization of ir-AM in cardiac myocytes and nonmyocytes
To investigate whether cultured rat ventricular myocytes and nonmyocytes secrete AM, the ir-AM concentration in the medium of cells cultured without FCS was measured. Both myocytes and nonmyocytes secreted ir-AM into serum-free medium in a time-dependent manner (Table 1Go). The basal release of ir-AM from myocytes per 105 cells was almost equivalent to that of nonmyocytes for 6–48 h of incubation.


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  		Table 1. Basal secretion of ir-AM from cultured cardiac myocytes and nonmyocytes

 
Ir-AM secreted from cultured cells was characterized by reverse phase HPLC. The ir-AM in the culture medium of cardiac myocytes consisted of one major and some minor components, and the major component was eluted at a retention time identical to that of authentic rat AM (Fig. 1Go). The elution profile of ir-AM in the medium of nonmyocytes also had some peaks similar to those of myocytes (data not shown).



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  		Figure 1. Reverse phase HPLC analysis of ir-AM in the culture medium of cardiac myocytes. A linear gradient elution of acetonitrile was 10–60% in 0.1% trifluoroacetic acid for 60 min at a flow rate of 1 ml/min, and 1-ml fractions were collected for RIA. The arrow indicates the elution position of synthetic rat AM.

 
Effects of IL-1ß and TNF{alpha} on the secretion of ir-AM in cardiac myocytes and nonmyocytes
In cardiac myocytes, the secretion level of ir-AM was significantly elevated only 48 h after stimulation with 0.1–10 ng/ml IL-1ß (Figs. 2AGo and 3AGo). The ir-AM secretion from myocytes was not increased by TNF{alpha} (Figs. 2AGo and 4AGo). While in cardiac nonmyocytes, both 10 ng/ml IL-1ß and 20 ng/ml TNF{alpha} remarkably stimulated the release of ir-AM after 6- to 48-h incubation (Fig. 2BGo). The stimulatory effect of IL-1ß on its release was concentration dependent (Fig. 3BGo). The maximal increase in ir-AM 48 h after stimulation with 100 ng/ml IL-1ß was about 13-fold. TNF{alpha} also stimulated the secretion of ir-AM in a concentration-dependent manner (Fig. 4BGo). Its maximal increase by 20–200 ng/ml TNF{alpha} was about 4-fold.



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  		Figure 2. Time course of the effects of IL-1ß (10 ng/ml; •) and TNF{alpha} (20 ng/ml; {blacksquare}) on the secretion of ir-AM in cultured cardiac myocytes (A) and nonmyocytes (B). Values are given as the mean ± SD of three measurements. Control ({circ}) indicates basal secretion. *, P < 0.05; ***, P < 0.001 (compared with the control).

 


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  		Figure 3. Dose-dependent effect of IL-1ß on the secretion of ir-AM after 24 h (•) or 48 h ({blacksquare}) of incubation in cultured cardiac myocytes (A) and nonmyocytes (B). Values are given as the mean ± SD of three measurements. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (compared with vehicle alone).

 


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  		Figure 4. Dose-dependent effect of TNF{alpha} on the secretion of ir-AM after 24 h (•) or 48 h ({blacksquare}) of incubation in cultured cardiac myocytes (A) and nonmyocytes (B). Values are given as the mean ± SD of three measurements. ***, P < 0.001 (compared with vehicle alone).

 
Expression of AM mRNA in cardiac myocytes and nonmyocytes and effects of IL-1ß and TNF{alpha} on its level
The expression of rat AM mRNA in cultured rat cardiac myocytes and nonmyocytes was examined by Northern blot analysis. As shown in Fig. 5Go, a single band hybridizing to the rat AM cDNA probe was found in both myocytes and nonmyocytes. Figure 6Go shows a quantitative analysis of these blots corrected for the levels of GAPDH mRNA as an internal standard. The expression of AM mRNA in myocytes was increased by IL-1ß after 24 h (1.6-fold) and 48 h (2.7-fold) and was slightly, but significantly, increased by TNF{alpha} after 24 h (1.3-fold). In nonmyocytes, the AM mRNA level was remarkably elevated by IL-1ß (24 h, 6.3-fold; 48 h, 14.2-fold) and by TNF{alpha} (24 h, 4.3-fold; 48 h, 6.8-fold).



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  		Figure 5. Representative image of the expression of rat AM, ANP, and GAPDH mRNA in cultured cardiac myocytes and nonmyocytes with or without treatment with IL-1ß (10 ng/ml) or TNF{alpha} (20 ng/ml).

 


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  		Figure 6. Quantitative analysis of AM transcripts in cultured cardiac myocytes (A) and nonmyocytes (B) with or without treatment with IL-1ß or TNF{alpha}. Open bars, Control; solid bars, IL-1ß (10 ng/ml); hatched bars, TNF{alpha} (20 ng/ml). Values shown were corrected by using the density of the corresponding GAPDH mRNA. Data represent the mean ± SD of three measurements. Control indicates vehicle alone. *, P < 0.05; **, P < 0.01; ***, P < 0.001 (compared with the control).

 
Effects of IL-1ß and TNF{alpha} on the secretion of ir-ANP and the expression of ANP mRNA in cardiac myocytes
As shown in Table 2Go and Fig. 5Go, cultured neonatal rat cardiac myocytes secreted a high amount of ir-ANP, and strong gene expression of ANP was observed in those cells. Neither IL-1ß nor TNF{alpha} had any stimulatory effect on the release of ir-ANP or on the gene expression of ANP in cardiac myocytes, except a slight increase in the mRNA level 48 h after treatment with IL-1ß (Table 2Go).


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  		Table 2. Effects of IL-1ß and TNF{alpha} on ir-ANP secretion and its mRNA level in cultured cardiac myocytes

 
Effects of angiotensin II, endothelin-1, and phenylephrine on the secretion of ir-AM and ir-ANP in cardiac myocytes and nonmyocytes
We further examined the effects of angiotensin II, endothelin-1, and phenylephrine, which are factors also known to be involved in cardiac hypertrophy and heart failure, on the release of ir-AM and ir-ANP from cultured ventricular cells. None of these agents stimulated the secretion of ir-AM in myocytes and nonmyocytes, whereas ir-ANP secretion in myocytes was significantly increased by 10-7-10-6 M endothelin-1 and 10-5-10-4 M phenylephrine (Table 3Go).


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  		Table 3. Effects of angiotensin II, endothelin-1, and phenylephrine on ir-AM and ir-ANP secretion in cultured cardiac myocytes and nonmyocytes

 

   Discussion
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
References
 
The present study has demonstrated for the first time that both purified cardiac ventricular myocytes and nonmyocytes actively release AM and express the AM gene. In basal conditions, the secretion rates of AM from myocytes and nonmyocytes were almost equivalent. The amount of ir-AM secreted from cultured myocytes or nonmyocytes (~20 fmol/105 cells·24 h) was comparable to that from other cultured cells, such as vascular smooth muscle cells (~10 fmol/105 cells·24 h) or vascular endothelial cells (~60 fmol/105 cells·24 h), as previously described by Sugo et al. (2, 3). Therefore, the present data indicate that neonatal rat ventricular cardiocytes release a considerable amount of AM.

We have clearly demonstrated that IL-1ß and TNF{alpha} stimulate the production and secretion of AM in cultured ventricular cells. It has been reported that ventricular levels as well as plasma concentrations of AM are increased in human congestive heart failure (9, 18). We and another group have shown that volume overload and pressure overload stimulate AM gene expression in the rat heart (10, 19). As circulating IL-1 and TNF levels are known to be elevated in congestive heart failure (12, 20, 21, 22), these observations and our results suggest that the high levels of plasma IL-1 and TNF in heart failure may partly contribute to the accelerated production of AM from the failing heart.

In the present study, stimulation by IL-1ß and TNF{alpha} of AM production and secretion was stronger in nonmyocytes (cardiac fibroblasts) than in myocytes. Both IL-1ß and TNF{alpha} have been shown to induce cellular hypertrophy in cardiac myocytes (13, 14). On the other hand, the cross-talk between cardiac fibroblasts and myocytes in the process of myocyte cell hypertrophy has been examined, and some reports have shown that cardiac fibroblasts may play a critical role in mediating the hypertrophic response to angiotensin II and endothelin in the heart (23, 24, 25). Although direct actions of AM on cultured cardiomyocytes were not elucidated in our study, cardiac myocytes have been revealed to have specific binding sites for AM (26), and a recent paper has reported that AM suppresses the protein synthesis of cultured cardiomyocytes (27). On the basis of these observations, AM released from fibroblasts with strong stimulation by IL-1ß and TNF{alpha} may participate in myocyte cell hypertrophy as a paracrine modulator.

The present study has shown that neither IL-1ß nor TNF{alpha} stimulates ANP release in ventricular myocytes. By contrast, agents such as endothelin and phenylephrine stimulated the secretion of ANP from cultured ventricular myocytes, compatible with the previous observations (15, 28), but the secretion of AM in myocytes and nonmyocytes was not stimulated by these agents. Therefore, it is possible that the production of AM in cardiac myocytes is regulated differently from that of ANP by several humoral factors that are involved in cardiac hypertrophy and heart failure, such as catecholamine, endothelin, and cytokines.

The pathophysiological roles of AM released from ventricular cardiocytes augmented by IL-1ß and TNF{alpha} are currently unclear. Several cytokines, including IL-1ß and TNF{alpha}, are known to depress myocardial contractility in part via inducible nitric oxide synthase activation (29, 30, 31, 32). Recently, Ikeda et al. (33) revealed that AM augments inducible nitric oxide synthase expression in IL-1ß-stimulated cardiac myocytes. Ikenouchi et al. (34) have shown that AM has a negative inotropic effect on isolated rabbit ventricular cells. Taken together, AM may act as a endogenous enhancer of nitric oxide production and be involved in the reduction of myocardial contractility by cytokines. On the other hand, some reports (35, 36) have suggested that IL-1 and TNF may have beneficial effects on the heart during times of ischemic stress. In addition, AM has been shown to elicit the beneficial hemodynamic effect in sheep with heart failure (37). Further investigations are essential to clarify the physiological and pathophysiological significance of AM in cardiac myocytes.


   Acknowledgments
 
We thank Yoshihiko Saito, Masahiro Ishikawa, and Kazuwa Nakao (Department of Medicine and Clinical Science, Kyoto University Graduate School of Medicine, Kyoto, Japan) for their helpful advice on the technique for cell culture. We also thank Ms. Yoko Saito for her technical assistance.


   Footnotes
 
1 This work was supported in part by Special Coordination Funds for Promoting Science and Technology (Encouragement System of COE) from the Science and Technology Agency of Japan, the Ministry of Health and Welfare, and the Human Science Foundation of Japan. Back

Received April 7, 1998.


   References
Top
 Abstract
 Introduction
 Materials and Methods
 Results
 Discussion
 References
 

  1. Kitamura K, Kangawa K, Kawamoto M, Ichiki Y, Nakamura S, Matsuo H, Eto T 1993 Adrenomedullin: a novel hypotensive peptide isolated from human pheochromocytoma. Biochem Biophys Res Commun 192:553–560[CrossRef][Medline]
  2. Sugo S, Minamino N, Kangawa K, Miyamoto K, Kitamura K, Sakata J, Eto T, Matsuo H 1994 Endothelial cells actively synthesize and secrete adrenomedullin. Biochem Biophys Res Commun 201:1160–1166[CrossRef][Medline]
  3. Sugo S, Minamino N, Shoji H, Kangawa K, Kitamura K, Eto T, Matsuo H 1994 Production and secretion of adrenomedullin from vascular smooth muscle cells: augmented production by tumor necrosis factor-{alpha}. Biochem Biophys Res Commun 203:719–726[CrossRef][Medline]
  4. Horio T, Kohno M, Kano H, Ikeda M, Yasunari K, Yokokawa K, Minami M, Takeda T 1995 Adrenomedullin as a novel antimigration factor of vascular smooth muscle cells. Circ Res 77:660–664[Abstract/Free Full Text]
  5. Kano H, Kohno M, Yasunari K, Yokokawa K, Horio T, Ikeda M, Minami M, Hanehira T, Takeda T, Yoshikawa J 1996 Adrenomedullin as a novel antiproliferative factor of vascular smooth muscle cells. J Hypertens 14:209–213[CrossRef][Medline]
  6. Kato H, Shichiri M, Marumo F, Hirata Y 1997 Adrenomedullin as an autocrine/paracrine apoptosis survival factor for rat endothelial cells. Endocrinology 138:2615–2620[Abstract/Free Full Text]
  7. Sakata J, Shimokubo T, Kitamura K, Nishizono M, Ichiki Y, Kangawa K, Matsuo H, Eto T 1994 Distribution and characterization of immunoreactive rat adrenomedullin in tissue and plasma. FEBS Lett 352:105–108[CrossRef][Medline]
  8. Jougasaki M, Wei CM, Heublein DM, Sandberg SM, Burnett Jr JC 1995 Immunohistochemical localization of adrenomedullin in canine heart and aorta. Peptides 16:773–775[CrossRef][Medline]
  9. Jougasaki M, Wei CM, McKinley LJ, Burnett Jr JC 1995 Elevation of circulating and ventricular adrenomedullin in human congestive heart failure. Circulation 92:286–289[Abstract/Free Full Text]
  10. Nishikimi T, Horio T, Sasaki T, Yoshihara F, Takishita S, Miyata A, Matsuo H, Kangawa K 1997 Cardiac production and secretion of adrenomedullin are increased in heart failure. Hypertension 30:1369–1375[Abstract/Free Full Text]
  11. Sugo S, Minamino N, Shoji H, Kangawa K, Kitamura K, Eto T, Matsuo H 1995 Interleukin-1, tumor necrosis factor and lipopolysaccharide additively stimulate production of adrenomedullin in vascular smooth muscle cells. Biochem Biophys Res Commun 207:25–32[CrossRef][Medline]
  12. Testa M, Yeh M, Lee P, Fanelli R, Loperfido F, Berman JW, LeJemtel TH 1996 Circulating levels of cytokines and their endogenous modulators in patients with mild to severe congestive heart failure due to coronary artery disease or hypertension. J Am Coll Cardiol 28:964–971[Abstract]
  13. Thaik CM, Calderone A, Takahashi N, Colucci WS 1995 Interleukin-1ß modulates the growth and phenotype of neonatal rat cardiac myocytes. J Clin Invest 96:1093–1099
  14. Yokoyama T, Nakano M, Bednarczyk JL, McIntyre BW, Entman M, Mann DL 1997 Tumor necrosis factor-{alpha} provokes a hypertrophic growth response in adult cardiac myocytes. Circulation 95:1247–1252[Abstract/Free Full Text]
  15. Nakagawa O, Ogawa Y, Itoh H, Suga S, Komatsu Y, Kishimoto I, Nishino K, Yoshimasa T, Nakao K 1995 Rapid transcriptional activation and early mRNA turnover of brain natriuretic peptide in cardiocyte hypertrophy. J Clin Invest 96:1280–1287
  16. Villarreal FJ, Kim NN, Ungab GD, Printz MP, Dillmann WH 1993 Identification of functional angiotensin II receptors on rat cardiac fibroblasts. Circulation 88:2849–2861[Abstract/Free Full Text]
  17. Aburaya M, Minamino N, Hino J, Kangawa K, Matsuo H 1989 Distribution and molecular forms of brain natriuretic peptide in the central nervous system, heart and peripheral tissue of rat. Biochem Biophys Res Commun 165:880–887[CrossRef][Medline]
  18. Jougasaki M, Rodeheffer RJ, Redfield MM, Yamamoto K, Wei CM, McKinley LJ, Burnett Jr JC 1996 Cardiac secretion of adrenomedullin in human heart failure. J Clin Invest 97:2370–2376[Medline]
  19. Romppanen H, Marttila M, Magga J, Vuolteenaho O, Kinnunen P, Szokodi I, Ruskoaho H 1997 Adrenomedullin gene expression in the rat heart is stimulated by acute pressure overload: blunted effect in experimental hypertension. Endocrinology 138:2636–2639[Abstract/Free Full Text]
  20. Levine B, Kalman J, Mayer L, Fillit HM, Packer M 1990 Elevated circulating levels of tumor necrosis factor in severe chronic heart failure. N Engl J Med 323:236–241[Abstract]
  21. Dutka DP, Elborn JS, Delamere F, Shale DJ, Morris GK 1993 Tumor necrosis factor {alpha} in severe congestive cardiac failure. Br Heart J 70:141–143[Abstract/Free Full Text]
  22. Milani RV, Mehra MR, Endres S, Eigler A, Cooper ES, Lavie Jr CJ, Ventura HO 1996 The clinical relevance of circulating tumor necrosis factor-{alpha} in acute decompensated chronic heart failure without cachexia. Chest 110:992–995[Abstract/Free Full Text]
  23. Kim NN, Villarreal FJ, Printz MP, Lee AA, Dillmann WH 1995 Trophic effects of angiotensin II on neonatal rat cardiac myocytes are mediated by cardiac fibroblasts. Am J Physiol 269:E426–E437
  24. Sil P, Sen S 1997 Angiotensin II and myocyte growth: role of fibroblasts. Hypertension 30:209–216[Abstract/Free Full Text]
  25. Harada M, Itoh H, Nakagawa O, Ogawa Y, Miyamoto Y, Kuwahara K, Ogawa E, Igaki T, Yamashita J, Masuda I, Yoshimasa T, Tanaka I, Saito Y, Nakao K 1997 Significance of ventricular myocytes and nonmyocytes interaction during cardiocyte hypertrophy: evidence for endothelin-1 as a paracrine hypertrophic factor from cardiac nonmyocytes. Circulation 96:3737–3744[Abstract/Free Full Text]
  26. Owji AA, Smith DM, Coppock HA, Morgan DGA, Bhogal R, Ghatei MA, Bloon SR 1995 An abundant and specific binding site for the novel vasodilator adrenomedullin in the rat. Endocrinology 136:2127–2134[Abstract]
  27. Tsuruda T, Kato J, Kitamura K, Kuwasako K, Imamura T, Koiwaya Y, Tsuji T, Kangawa K, Eto T 1998 Adrenomedullin: a possible autocrine or paracrine inhibitor of hypertrophy of cardiomyocytes. Hypertension 31:505–510[Abstract/Free Full Text]
  28. Sei CA, Irons CE, Sprenkle AB, McDonough PM, Brown JH, Glembotski CC 1991 The {alpha}-adrenergic stimulation of atrial natriuretic factor expression in cardiac myocytes requires calcium influx, protein kinase C, and calmodulin-regulated pathways. J Biol Chem 266:15910–15916[Abstract/Free Full Text]
  29. Finkel MS, Oddis CV, Jacob TD, Watkins SC, Hattler BG, Simmons RL 1992 Negative inotropic effects of cytokines on the heart mediated by nitric oxide. Science 257:387–389[Abstract/Free Full Text]
  30. Evans HG, Lewis MJ, Shah AM 1993 Interleukin-1ß modulates myocardial contraction via dexamethasone sensitive production of nitric oxide. Cardiovasc Res 27:1486–1490[Abstract/Free Full Text]
  31. Ungureanu-Longrois D, Balligand JL, Simmons WW, Okada I, Kobzik L, Lowenstein CJ, Kunkel SL, Michel T, Kelly RA, Smith TW 1995 Induction of nitric oxide synthase activity by cytokines in ventricular myocytes is necessary but not sufficient to decrease contractile responsiveness to ß-adrenergic agonists. Circ Res 77:494–502[Abstract/Free Full Text]
  32. Stein B, Frank P, Schmitz W, Scholz H, Thoenes M 1996 Endotoxin and cytokines induce direct cardiodepressive effects in mammalian cardiomyocytes via induction of nitric oxide synthase. J Mol Cell Cardiol 28:1631–1639[CrossRef][Medline]
  33. Ikeda U, Kanbe T, Kawahara Y, Yokoyama M, Shimada K 1996 Adrenomedullin augments inducible nitric oxide synthase expression in cytokine-stimulated cardiac myocytes. Circulation 94:2560–2565[Abstract/Free Full Text]
  34. Ikenouchi H, Kangawa K, Matsuo H, Hirata Y 1997 Negative inotropic effect of adrenomedullin in isolated adult rabbit cardiac ventricular myocytes. Circulation 95:2318–2324[Abstract/Free Full Text]
  35. Brown JM, White CW, Terada LS, Grosso MA, Shanley PF, Mulvin DW, Banerjee A, Whitman GJR, Harken AH, Repine JE 1990 Interleukin 1 pretreatment decreases ischemia/reperfusion injury. Proc Natl Acad Sci USA 87:5026–5030[Abstract/Free Full Text]
  36. Eddy LJ, Goeddel DV, Wong GHW 1992 Tumor necrosis factor-{alpha} pretreatment is protective in a rat model of myocardial ischemia-reperfusion injury. Biochem Biophys Res Commun 184:1056–1059[CrossRef][Medline]
  37. Rademaker MT, Charles CJ, Lewis LK, Yandle TG, Cooper GJS, Coy DH, Richards AM, Nicholls MG 1997 Beneficial hemodynamic and renal effects of adrenomedullin in an ovine model of heart failure. Circulation 96:1983–1990[Abstract/Free Full Text]



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J. Gerontol. A Biol. Sci. Med. Sci., January 1, 2007; 62(1): 41 - 49.
[Abstract] [Full Text] [PDF]

Home page
 J. Leukoc. Biol.Home page
E. Zudaire, S. Portal-Nunez, and F. Cuttitta
The central role of adrenomedullin in host defense
J. Leukoc. Biol., August 1, 2006; 80(2): 237 - 244.
[Abstract] [Full Text] [PDF]

Home page
 J. Pharmacol. Exp. Ther.Home page
Y. Zhao, D. Bell, L. R. Smith, L. Zhao, A. B. Devine, E. M. McHenry, D. P. Nicholls, and B. J. McDermott
Differential Expression of Components of the Cardiomyocyte Adrenomedullin/Intermedin Receptor System following Blood Pressure Reduction in Nitric Oxide-Deficient Hypertension
J. Pharmacol. Exp. Ther., March 1, 2006; 316(3): 1269 - 1281.
[Abstract] [Full Text] [PDF]

Home page
 Mol Hum ReprodHome page
A. Al-Ghafra, N.M. Gude, S.P. Brennecke, and R.G. King
Increased adrenomedullin protein content and mRNA expression in human fetal membranes but not placental tissue in pre-eclampsia
Mol. Hum. Reprod., March 1, 2006; 12(3): 181 - 186.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
D. Bell, Y.-Y. Zhao, E. J. Kelso, E. M. McHenry, L. M. Rush, V. M. Lamont, D. P. Nicholls, and B. J. McDermott
Upregulation of adrenomedullin and its receptor components during cardiomyocyte hypertrophy induced by chronic inhibition of nitric oxide synthesis in rats
Am J Physiol Heart Circ Physiol, February 1, 2006; 290(2): H904 - H914.
[Abstract] [Full Text] [PDF]

Home page
 Cardiovasc ResHome page
T. Nishikimi, K. Tadokoro, K. Akimoto, Y. Mori, Y. Ishikawa, K. Ishimura, T. Horio, K. Kangawa, and H. Matsuoka
Response of adrenomedullin system to cytokine in cardiac fibroblasts-role of adrenomedullin as an antifibrotic factor
Cardiovasc Res, April 1, 2005; 66(1): 104 - 113.
[Abstract] [Full Text] [PDF]

Home page
 J Am Coll CardiolHome page
T. Soeki, I. Kishimoto, H. Okumura, T. Tokudome, T. Horio, K. Mori, and K. Kangawa
C-type natriuretic peptide, a novel antifibrotic and antihypertrophic agent, prevents cardiac remodeling after myocardial infarction
J. Am. Coll. Cardiol., February 15, 2005; 45(4): 608 - 616.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
T. Tokudome, T. Horio, M. Fukunaga, H. Okumura, J. Hino, K. Mori, F. Yoshihara, S.-I. Suga, Y. Kawano, M. Kohno, et al.
Ventricular Nonmyocytes Inhibit Doxorubicin-Induced Myocyte Apoptosis: Involvement of Endogenous Endothelin-1 as a Paracrine Factor
Endocrinology, May 1, 2004; 145(5): 2458 - 2466.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
T. Tokudome, T. Horio, T. Soeki, K. Mori, I. Kishimoto, S.-i. Suga, F. Yoshihara, Y. Kawano, M. Kohno, and K. Kangawa
Inhibitory Effect of C-Type Natriuretic Peptide (CNP) on Cultured Cardiac Myocyte Hypertrophy: Interference between CNP and Endothelin-1 Signaling Pathways
Endocrinology, May 1, 2004; 145(5): 2131 - 2140.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
S. Mittra, J.-M. Hyvelin, Q. Shan, F. Tang, and J.-P. Bourreau
Role of cyclooxygenase in ventricular effects of adrenomedullin: is adrenomedullin a double-edged sword in sepsis?
Am J Physiol Heart Circ Physiol, March 1, 2004; 286(3): H1034 - H1042.
[Abstract] [Full Text] [PDF]

Home page
 EndocrinologyHome page
T. Horio, T. Tokudome, T. Maki, F. Yoshihara, S.-i. Suga, T. Nishikimi, M. Kojima, Y. Kawano, and K. Kangawa
Gene Expression, Secretion, and Autocrine Action of C-Type Natriuretic Peptide in Cultured Adult Rat Cardiac Fibroblasts
Endocrinology, June 1, 2003; 144(6): 2279 - 2284.
[Abstract] [Full Text] [PDF]

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 EndocrinologyHome page
T. Tokudome, T. Horio, F. Yoshihara, S.-i. Suga, Y. Kawano, M. Kohno, and K. Kangawa
Adrenomedullin Inhibits Doxorubicin-Induced Cultured Rat Cardiac Myocyte Apoptosis via a cAMP-Dependent Mechanism
Endocrinology, September 1, 2002; 143(9): 3515 - 3521.
[Abstract] [Full Text] [PDF]

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 FASEB J.Home page
N. S. WAYMAN, Y. HATTORI, M. C. MCDONALD, H. MOTA-FILIPE, S. CUZZOCREA, B. PISANO, P. K. CHATTERJEE, and C. THIEMERMANN
Ligands of the peroxisome proliferator-activated receptors (PPAR-{gamma} and PPAR-{alpha}) reduce myocardial infarct size
FASEB J, July 1, 2002; 16(9): 1027 - 1040.
[Abstract] [Full Text] [PDF]

Home page
 J Am Coll CardiolHome page
F. Yoshihara, T. Nishikimi, Y. Sasako, J. Hino, J. Kobayashi, K. Minatoya, K. Bando, Y. Kosakai, T. Horio, S.-i. Suga, et al.
Plasma atrial natriuretic peptide concentration inversely correlates with left atrial collagen volume fraction in patients with atrial fibrillation: Plasma ANP as a possible biochemical marker to predict the outcome of the maze procedure
J. Am. Coll. Cardiol., January 16, 2002; 39(2): 288 - 294.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
T. Nishikimi, F. Yoshihara, A. Kanazawa, I. Okano, T. Horio, N. Nagaya, C. Yutani, H. Matsuo, H. Matsuoka, and K. Kangawa
Role of increased circulating and renal adrenomedullin in rats with malignant hypertension
Am J Physiol Regulatory Integrative Comp Physiol, December 1, 2001; 281(6): R2079 - R2087.
[Abstract] [Full Text] [PDF]

Home page
 J Am Coll CardiolHome page
M.-W. Hwang, A. Matsumori, Y. Furukawa, K. Ono, M. Okada, A. Iwasaki, M. Hara, T. Miyamoto, M. Touma, and S. Sasayama
Neutralization of interleukin-1{beta} in the acute phase of myocardial infarction promotes the progression of left ventricular remodeling
J. Am. Coll. Cardiol., November 1, 2001; 38(5): 1546 - 1553.
[Abstract] [Full Text] [PDF]

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 Cardiovasc ResHome page
M. Jougasaki and J. C Burnett Jr.
Continuing insights into the heart as an endocrine organ: adrenomedullin and cardiac fibroblasts
Cardiovasc Res, March 1, 2001; 49(4): 695 - 696.
[Full Text] [PDF]

Home page
 Cardiovasc ResHome page
Y. Tomoda, K. Kikumoto, Y. Isumi, T. Katafuchi, A. Tanaka, K. Kangawa, K. Dohi, and N. Minamino
Cardiac fibroblasts are major production and target cells of adrenomedullin in the heart in vitro
Cardiovasc Res, March 1, 2001; 49(4): 721 - 730.
[Abstract] [Full Text] [PDF]

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 HypertensionHome page
F. Yoshihara, T. Nishikimi, I. Okano, T. Horio, C. Yutani, H. Matsuo, S. Takishita, T. Ohe, and K. Kangawa
Alterations of Intrarenal Adrenomedullin and Its Receptor System in Heart Failure Rats
Hypertension, February 1, 2001; 37(2): 216 - 222.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Heart Circ. Physiol.Home page
T. Nishikimi, A. Miyata, T. Horio, F. Yoshihara, N. Nagaya, S. Takishita, C. Yutani, H. Matsuo, H. Matsuoka, and K. Kangawa
Urocortin, a member of the corticotropin-releasing factor family, in normal and diseased heart
Am J Physiol Heart Circ Physiol, December 1, 2000; 279(6): H3031 - H3039.
[Abstract] [Full Text] [PDF]

Home page
 Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
P. Kinnunen, I. Szokodi, M. G. Nicholls, and H. Ruskoaho
Impact of NO on ET-1- and AM-induced inotropic responses: potentiation by combined administration
Am J Physiol Regulatory Integrative Comp Physiol, August 1, 2000; 279(2): R569 - R575.
[Abstract] [Full Text] [PDF]

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 Clin. Cancer Res.Home page
S. Hague, L. Zhang, M. K. Oehler, S. Manek, I. Z. MacKenzie, R. Bicknell, and M. C. P. Rees
Expression of the Hypoxically Regulated Angiogenic Factor Adrenomedullin Correlates with Uterine Leiomyoma Vascular Density
Clin. Cancer Res., July 1, 2000; 6(7): 2808 - 2814.
[Abstract] [Full Text]

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 HypertensionHome page
T. Tsuruda, J. Kato, K. Kitamura, T. Imamura, Y. Koiwaya, K. Kangawa, I. Komuro, Y. Yazaki, and T. Eto
Enhanced Adrenomedullin Production by Mechanical Stretching in Cultured Rat Cardiomyocytes
Hypertension, June 1, 2000; 35(6): 1210 - 1214.
[Abstract] [Full Text] [PDF]

Home page
 Endocr. Rev.Home page
J. P. Hinson, S. Kapas, and D. M. Smith
Adrenomedullin, a Multifunctional Regulatory Peptide
Endocr. Rev., April 1, 2000; 21(2): 138 - 167.
[Abstract] [Full Text]

Home page
 Am. J. Physiol. Regul. Integr. Comp. Physiol.Home page
N. Nagaya, T. Nishikimi, F. Yoshihara, T. Horio, A. Morimoto, and K. Kangawa
Cardiac adrenomedullin gene expression and peptide accumulation after acute myocardial infarction in rats
Am J Physiol Regulatory Integrative Comp Physiol, April 1, 2000; 278(4): R1019 - R1026.
[Abstract] [Full Text] [PDF]

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 CirculationHome page
E. Oie, L. E. Vinge, A. Yndestad, C. Sandberg, H. K. Grogaard, and H. Attramadal
Induction of a Myocardial Adrenomedullin Signaling System During Ischemic Heart Failure in Rats
Circulation, February 1, 2000; 101(4): 415 - 422.
[Abstract] [Full Text] [PDF]

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 HypertensionHome page
T. Horio, T. Nishikimi, F. Yoshihara, H. Matsuo, S. Takishita, and K. Kangawa
Inhibitory Regulation of Hypertrophy by Endogenous Atrial Natriuretic Peptide in Cultured Cardiac