Skip to main content
NCBI home page
The Journal of Clinical Investigation logoLink to The Journal of Clinical Investigation
. 1996 Jan 15;97(2):366–372. doi: 10.1172/JCI118424

Endothelin-3 induces hypertrophy of cardiomyocytes by the endogenous endothelin-1-mediated mechanism.

M Tamamori 1, H Ito 1, S Adachi 1, H Akimoto 1, F Marumo 1, M Hiroe 1
PMCID: PMC507026  PMID: 8567956

Abstract

We have recently reported that endothelin-1 (ET-1) mediates angiotensin II-induced hypertrophy of cardiomyocytes as an autocrine/paracrine factor. In the present study, we examined whether endothelin-3 (ET-3) induces hypertrophy of cultured neonatal rat cardiomyocytes and whether endogenous ET-1 mediates this effect. ET-3 (10(-7) M) increased the cell surface area of cardiomyocytes after 48 h. ET-3 dose dependently (10(-9)-10(-7) M) stimulated protein synthesis as evaluated by [3H]leucine incorporation; the maximum response was 1.4-fold increase over the control at 10(-7) M. Since the response of cardiac hypertrophy is characterized by enhanced expression of fetal isoforms of muscle specific genes, the effect of ET-3 on steady state levels of mRNA for skeletal alpha-actin was evaluated by Northern blot analysis. ET-3 (10(-9)-10(-7) M) increased mRNA level for skeletal alpha-actin with a maximum response after 6 h. ET-3-induced [3H]leucine incorporation, skeletal alpha-actin mRNA and cell surface area were inhibited by a synthetic ETB receptor antagonist (BQ788). Interestingly, ET-3-induced skeletal alpha-actin gene expression and [3H]leucine incorporation were inhibited by a synthetic ETA receptor antagonist (BQ123) as well as by antisense oligonucleotides against peproET-1 mRNA. ET-3 (10(-7) M) transiently increased mRNA levels for ET-1 peaking at 30 min and stimulated the release of immunoreactive ET-1 from cardiomyocytes. These results suggest that endogenous ET-1 locally generated and secreted by cardiomyocytes may contribute to ET-3-induced cardiac hypertrophy as an autocrine/paracrine factor.

Full Text

The Full Text of this article is available as a PDF (404.5 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bishopric N. H., Simpson P. C., Ordahl C. P. Induction of the skeletal alpha-actin gene in alpha 1-adrenoceptor-mediated hypertrophy of rat cardiac myocytes. J Clin Invest. 1987 Oct;80(4):1194–1199. doi: 10.1172/JCI113179. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Chomczynski P., Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  3. Eguchi S., Hirata Y., Ihara M., Yano M., Marumo F. A novel ETA antagonist (BQ-123) inhibits endothelin-1-induced phosphoinositide breakdown and DNA synthesis in rat vascular smooth muscle cells. FEBS Lett. 1992 May 18;302(3):243–246. doi: 10.1016/0014-5793(92)80451-l. [DOI] [PubMed] [Google Scholar]
  4. Feinberg A. P., Vogelstein B. A technique for radiolabeling DNA restriction endonuclease fragments to high specific activity. Anal Biochem. 1983 Jul 1;132(1):6–13. doi: 10.1016/0003-2697(83)90418-9. [DOI] [PubMed] [Google Scholar]
  5. Hirata Y., Emori T., Eguchi S., Kanno K., Imai T., Ohta K., Marumo F. Endothelin receptor subtype B mediates synthesis of nitric oxide by cultured bovine endothelial cells. J Clin Invest. 1993 Apr;91(4):1367–1373. doi: 10.1172/JCI116338. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Ihara M., Noguchi K., Saeki T., Fukuroda T., Tsuchida S., Kimura S., Fukami T., Ishikawa K., Nishikibe M., Yano M. Biological profiles of highly potent novel endothelin antagonists selective for the ETA receptor. Life Sci. 1992;50(4):247–255. doi: 10.1016/0024-3205(92)90331-i. [DOI] [PubMed] [Google Scholar]
  7. Inoue A., Yanagisawa M., Kimura S., Kasuya Y., Miyauchi T., Goto K., Masaki T. The human endothelin family: three structurally and pharmacologically distinct isopeptides predicted by three separate genes. Proc Natl Acad Sci U S A. 1989 Apr;86(8):2863–2867. doi: 10.1073/pnas.86.8.2863. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Inoue A., Yanagisawa M., Takuwa Y., Mitsui Y., Kobayashi M., Masaki T. The human preproendothelin-1 gene. Complete nucleotide sequence and regulation of expression. J Biol Chem. 1989 Sep 5;264(25):14954–14959. [PubMed] [Google Scholar]
  9. Ishikawa K., Ihara M., Noguchi K., Mase T., Mino N., Saeki T., Fukuroda T., Fukami T., Ozaki S., Nagase T. Biochemical and pharmacological profile of a potent and selective endothelin B-receptor antagonist, BQ-788. Proc Natl Acad Sci U S A. 1994 May 24;91(11):4892–4896. doi: 10.1073/pnas.91.11.4892. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Ishikawa T., Li L. M., Shinmi O., Kimura S., Yanagisawa M., Goto K., Masaki T. Characteristics of binding of endothelin-1 and endothelin-3 to rat hearts. Developmental changes in mechanical responses and receptor subtypes. Circ Res. 1991 Oct;69(4):918–926. doi: 10.1161/01.res.69.4.918. [DOI] [PubMed] [Google Scholar]
  11. Ito H., Hirata Y., Adachi S., Tanaka M., Tsujino M., Koike A., Nogami A., Murumo F., Hiroe M. Endothelin-1 is an autocrine/paracrine factor in the mechanism of angiotensin II-induced hypertrophy in cultured rat cardiomyocytes. J Clin Invest. 1993 Jul;92(1):398–403. doi: 10.1172/JCI116579. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Ito H., Hirata Y., Hiroe M., Tsujino M., Adachi S., Takamoto T., Nitta M., Taniguchi K., Marumo F. Endothelin-1 induces hypertrophy with enhanced expression of muscle-specific genes in cultured neonatal rat cardiomyocytes. Circ Res. 1991 Jul;69(1):209–215. doi: 10.1161/01.res.69.1.209. [DOI] [PubMed] [Google Scholar]
  13. Ito H., Hiroe M., Hirata Y., Fujisaki H., Adachi S., Akimoto H., Ohta Y., Marumo F. Endothelin ETA receptor antagonist blocks cardiac hypertrophy provoked by hemodynamic overload. Circulation. 1994 May;89(5):2198–2203. doi: 10.1161/01.cir.89.5.2198. [DOI] [PubMed] [Google Scholar]
  14. Ito H., Hiroe M., Hirata Y., Tsujino M., Adachi S., Shichiri M., Koike A., Nogami A., Marumo F. Insulin-like growth factor-I induces hypertrophy with enhanced expression of muscle specific genes in cultured rat cardiomyocytes. Circulation. 1993 May;87(5):1715–1721. doi: 10.1161/01.cir.87.5.1715. [DOI] [PubMed] [Google Scholar]
  15. Ito H., Miller S. C., Billingham M. E., Akimoto H., Torti S. V., Wade R., Gahlmann R., Lyons G., Kedes L., Torti F. M. Doxorubicin selectively inhibits muscle gene expression in cardiac muscle cells in vivo and in vitro. Proc Natl Acad Sci U S A. 1990 Jun;87(11):4275–4279. doi: 10.1073/pnas.87.11.4275. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Kanno K., Hirata Y., Tsujino M., Imai T., Shichiri M., Ito H., Marumo F. Up-regulation of ETB receptor subtype mRNA by angiotensin II in rat cardiomyocytes. Biochem Biophys Res Commun. 1993 Aug 16;194(3):1282–1287. doi: 10.1006/bbrc.1993.1962. [DOI] [PubMed] [Google Scholar]
  17. Lee H. R., Henderson S. A., Reynolds R., Dunnmon P., Yuan D., Chien K. R. Alpha 1-adrenergic stimulation of cardiac gene transcription in neonatal rat myocardial cells. Effects on myosin light chain-2 gene expression. J Biol Chem. 1988 May 25;263(15):7352–7358. [PubMed] [Google Scholar]
  18. Miyauchi T., Yorikane R., Sakai S., Sakurai T., Okada M., Nishikibe M., Yano M., Yamaguchi I., Sugishita Y., Goto K. Contribution of endogenous endothelin-1 to the progression of cardiopulmonary alterations in rats with monocrotaline-induced pulmonary hypertension. Circ Res. 1993 Nov;73(5):887–897. doi: 10.1161/01.res.73.5.887. [DOI] [PubMed] [Google Scholar]
  19. Ohta K., Hirata Y., Shichiri M., Kanno K., Emori T., Tomita K., Marumo F. Urinary excretion of endothelin-1 in normal subjects and patients with renal disease. Kidney Int. 1991 Feb;39(2):307–311. doi: 10.1038/ki.1991.38. [DOI] [PubMed] [Google Scholar]
  20. Schiaffino S., Samuel J. L., Sassoon D., Lompré A. M., Garner I., Marotte F., Buckingham M., Rappaport L., Schwartz K. Nonsynchronous accumulation of alpha-skeletal actin and beta-myosin heavy chain mRNAs during early stages of pressure-overload--induced cardiac hypertrophy demonstrated by in situ hybridization. Circ Res. 1989 May;64(5):937–948. doi: 10.1161/01.res.64.5.937. [DOI] [PubMed] [Google Scholar]
  21. Shubeita H. E., McDonough P. M., Harris A. N., Knowlton K. U., Glembotski C. C., Brown J. H., Chien K. R. Endothelin induction of inositol phospholipid hydrolysis, sarcomere assembly, and cardiac gene expression in ventricular myocytes. A paracrine mechanism for myocardial cell hypertrophy. J Biol Chem. 1990 Nov 25;265(33):20555–20562. [PubMed] [Google Scholar]
  22. Simpson P., Savion S. Differentiation of rat myocytes in single cell cultures with and without proliferating nonmyocardial cells. Cross-striations, ultrastructure, and chronotropic response to isoproterenol. Circ Res. 1982 Jan;50(1):101–116. doi: 10.1161/01.res.50.1.101. [DOI] [PubMed] [Google Scholar]
  23. Yanagisawa M., Kurihara H., Kimura S., Tomobe Y., Kobayashi M., Mitsui Y., Yazaki Y., Goto K., Masaki T. A novel potent vasoconstrictor peptide produced by vascular endothelial cells. Nature. 1988 Mar 31;332(6163):411–415. doi: 10.1038/332411a0. [DOI] [PubMed] [Google Scholar]
  24. Yokokawa K., Kohno M., Yasunari K., Murakawa K., Takeda T. Endothelin-3 regulates endothelin-1 production in cultured human endothelial cells. Hypertension. 1991 Sep;18(3):304–315. doi: 10.1161/01.hyp.18.3.304. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Clinical Investigation are provided here courtesy of American Society for Clinical Investigation

RESOURCES