title: Role of protein kinase C in the expression of endothelin converting enzyme-1.
abstract: Increased expression of endothelin converting enzyme-1 (ECE-1) is associated with diabetic nephropathy. The molecular mechanisms underlying this association, as yet unknown, possibly involve protein kinase C (PKC) pathways. In the present study, we examined the effects of high glucose and PKC activation on ECE-1 expression in primary human umbilical vein endothelial cells (HUVECs) and in HUVEC line (EA.hy926). Increasing glucose concentration, but not mannitol, from 5.5-22.2 mmol/liter for 3 d, enhanced prepro endothelin-1 (ET-1) mRNA expression, ET-1 levels, ECE-1 protein, and mRNA expressions by 7, 4, 20, and 2.6-fold, respectively. High glucose increased ECE-1 protein expression dose and time dependently. By Western blot analysis, PKC-beta1, -beta2, and -delta isoform levels were significantly increased relative to other isoforms when glucose level was increased. Treatment with Rottlerin, a PKC-delta isoform inhibitor, reduced significantly the glucose-induced ET-1 secretion, and ECE-1 protein expression, but (S)-13-[(dimethylamino)methyl]-10,11,14,15-tetrahydro-4,9:16,21-dimetheno 1H,1(3)H-dibenzo[e,k]pyrrolo[3,4-h] (1, 4, 3) oxadiaza-cyclohexadecene-1,3(2H)-dione or Gö6976, specific PKC-beta and -alpha inhibitors, respectively, did not. Overexpression of PKC-delta but not PKC-alpha or -beta1 isoforms by adenovirus vector containing the respective cDNA in HUVECs incubated with 5.5 mmol/liter glucose, increased in parallel PKC proteins, and glucose-induced endothein-1 and ECE-1 protein expression by 4- to 6-fold. These results show that enhanced ECE-1 expression induced by hyperglycemia is partly due to activation of the PKC-delta isoform. Thus, inhibition of this PKC isoform may prevent diabetes-related increase in ET-1.
authors.affiliation: Department of Medicine, Diabetes Research Unit, Hadassah Hospital, Ein Kerem, Mt Scopus, and the Hebrew University Medical School, Jerusalem, Israel. m_khamaisi@rambam.health.gov.il
authors.forename: Mogher
authors.initials: M
authors.lastname: Khamaisi
authors.forename: Rachel
authors.initials: R
authors.lastname: Dahan
authors.forename: Saher
authors.initials: S
authors.lastname: Hamed
authors.forename: Zaid
authors.initials: Z
authors.lastname: Abassi
authors.forename: Samuel N
authors.initials: SN
authors.lastname: Heyman
authors.forename: Itamar
authors.initials: I
authors.lastname: Raz
mesh_terms: D016282:Aspartic Acid Endopeptidases / Q000235:genetics* / Q000378:metabolism; D002478:Cells, Cultured; D003920:Diabetes Mellitus / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D019332:Endothelin-1 / Q000235:genetics / Q000378:metabolism; D000072396:Endothelin-Converting Enzymes; D016232:Endothelins / Q000235:genetics / Q000378:metabolism; D015971:Gene Expression Regulation, Enzymologic / Q000187:drug effects; D005947:Glucose / Q000494:pharmacology; D006801:Humans; D006943:Hyperglycemia / Q000201:enzymology / Q000235:genetics / Q000378:metabolism; D007527:Isoenzymes / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D008666:Metalloendopeptidases / Q000235:genetics* / Q000378:metabolism; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism / Q000502:physiology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology; D013379:Substrate Specificity
chemical_list: D019332:Endothelin-1; D016232:Endothelins; D007527:Isoenzymes; D047428:Protein Kinase Inhibitors; D011493:Protein Kinase C; D016282:Aspartic Acid Endopeptidases; D008666:Metalloendopeptidases; C000606923:ECE1 protein, human; D000072396:Endothelin-Converting Enzymes; D005947:Glucose