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Angiotensin converting enzyme and kininase-II-like activities in cultured valvular interstitial cells of the rat heart.
Cardiovasc Res. 1995 Jan; 29(1):57-64.CR

Abstract

OBJECTIVE

The function of angiotensin converting enzyme (ACE) at cell sites of high collagen turnover, such as heart valves, is uncertain. The aim of this study was to assess ACE and kininase-II-like activities and collagen turnover in cultured valvular interstitial cells of the adult rat heart.

METHODS

The valvular interstitial cell phenotype was determined by immunolabelling (rhodamine phalloidin, desmin, and Griffonia simplicifolia lectin), and the presence of ACE mRNA and protein was confirmed by reverse transcriptase-polymerase chain reaction analysis, ACE monoclonal antibody and in vitro autoradiography, respectively. ACE and kininase-II-like activities in valvular interstitial cells were analysed by high performance liquid chromatography. Angiotensin II (AT1) and bradykinin receptors in valvular interstitial cell membranes were examined by western immunoblotting and binding assay. Type I collagen and collagenase in valvular interstitial cell culture media were determined by ELISA and zymography, respectively. Type I collagen mRNA expression in cultured valvular interstitial cells was determined by northern blot analysis and in situ hybridisation.

RESULTS

In intact valvular interstitial cells or their cell membrane we found: (1) actin microfilaments, but not desmin or lectin labelling; (2) ACE mRNA expression and binding activity; (3) conversion of angiotensin I to angiotensin II, which was completely inhibited by 50 microM lisinopril, while kinase-II-like activity exceeded ACE activity and was not inhibited by lisinopril; (4) AT1 and bradykinin receptors in valvular interstitial cell membrane preparations; (5) type I collagen mRNA expression and collagenase activity; and (6) angiotensin II induced increase in type I collagen synthesis and mRNA expression.

CONCLUSIONS

Cultured valvular interstitial cells represent a nonendothelial, non-smooth-muscle cell type that expresses mRNA for ACE and type I collagen. ACE and kininase-II-like activities in valvular interstitial cells may be involved in the regulation of peptides that influence collagen turnover. Angiotensin II stimulates type I collagen synthesis and mRNA expression in these cells.

Authors+Show Affiliations

Department of Internal Medicine, University of Missouri Health Sciences Center, Columbia.No affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info availableNo affiliation info available

Pub Type(s)

Journal Article

Language

eng

PubMed ID

7895240

Citation

Katwa, L C., et al. "Angiotensin Converting Enzyme and kininase-II-like Activities in Cultured Valvular Interstitial Cells of the Rat Heart." Cardiovascular Research, vol. 29, no. 1, 1995, pp. 57-64.
Katwa LC, Ratajska A, Cleutjens JP, et al. Angiotensin converting enzyme and kininase-II-like activities in cultured valvular interstitial cells of the rat heart. Cardiovasc Res. 1995;29(1):57-64.
Katwa, L. C., Ratajska, A., Cleutjens, J. P., Sun, Y., Zhou, G., Lee, S. J., & Weber, K. T. (1995). Angiotensin converting enzyme and kininase-II-like activities in cultured valvular interstitial cells of the rat heart. Cardiovascular Research, 29(1), 57-64.
Katwa LC, et al. Angiotensin Converting Enzyme and kininase-II-like Activities in Cultured Valvular Interstitial Cells of the Rat Heart. Cardiovasc Res. 1995;29(1):57-64. PubMed PMID: 7895240.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Angiotensin converting enzyme and kininase-II-like activities in cultured valvular interstitial cells of the rat heart. AU - Katwa,L C, AU - Ratajska,A, AU - Cleutjens,J P, AU - Sun,Y, AU - Zhou,G, AU - Lee,S J, AU - Weber,K T, PY - 1995/1/1/pubmed PY - 1995/1/1/medline PY - 1995/1/1/entrez SP - 57 EP - 64 JF - Cardiovascular research JO - Cardiovasc Res VL - 29 IS - 1 N2 - OBJECTIVE: The function of angiotensin converting enzyme (ACE) at cell sites of high collagen turnover, such as heart valves, is uncertain. The aim of this study was to assess ACE and kininase-II-like activities and collagen turnover in cultured valvular interstitial cells of the adult rat heart. METHODS: The valvular interstitial cell phenotype was determined by immunolabelling (rhodamine phalloidin, desmin, and Griffonia simplicifolia lectin), and the presence of ACE mRNA and protein was confirmed by reverse transcriptase-polymerase chain reaction analysis, ACE monoclonal antibody and in vitro autoradiography, respectively. ACE and kininase-II-like activities in valvular interstitial cells were analysed by high performance liquid chromatography. Angiotensin II (AT1) and bradykinin receptors in valvular interstitial cell membranes were examined by western immunoblotting and binding assay. Type I collagen and collagenase in valvular interstitial cell culture media were determined by ELISA and zymography, respectively. Type I collagen mRNA expression in cultured valvular interstitial cells was determined by northern blot analysis and in situ hybridisation. RESULTS: In intact valvular interstitial cells or their cell membrane we found: (1) actin microfilaments, but not desmin or lectin labelling; (2) ACE mRNA expression and binding activity; (3) conversion of angiotensin I to angiotensin II, which was completely inhibited by 50 microM lisinopril, while kinase-II-like activity exceeded ACE activity and was not inhibited by lisinopril; (4) AT1 and bradykinin receptors in valvular interstitial cell membrane preparations; (5) type I collagen mRNA expression and collagenase activity; and (6) angiotensin II induced increase in type I collagen synthesis and mRNA expression. CONCLUSIONS: Cultured valvular interstitial cells represent a nonendothelial, non-smooth-muscle cell type that expresses mRNA for ACE and type I collagen. ACE and kininase-II-like activities in valvular interstitial cells may be involved in the regulation of peptides that influence collagen turnover. Angiotensin II stimulates type I collagen synthesis and mRNA expression in these cells. SN - 0008-6363 UR - https://www.unboundmedicine.com/medline/citation/7895240/Angiotensin_converting_enzyme_and_kininase_II_like_activities_in_cultured_valvular_interstitial_cells_of_the_rat_heart_ L2 - https://antibodies.cancer.gov/detail/CPTC-ATM-4 DB - PRIME DP - Unbound Medicine ER -