ADP-Ribosyl Cyclase in Rat Vascular Smooth Muscle Cells : Properties and Regulation

• Published in: Circulation research 2000-06, Vol.86 (11), p.1153-1159
• Main Authors: de Toledo, Frederico G. S, Cheng, Jingfei, Liang, Mingyu, Chini, Eduardo N, Dousa, Thomas P
• Format: Article
• Language: English
• Subjects: ADP-ribosyl Cyclase; ADP-ribosyl Cyclase 1; Animals; Antigens, CD; Antigens, Differentiation - metabolism; Biological and medical sciences; Blood vessels and receptors; Calcitriol - pharmacology; Cells, Cultured; Copper - pharmacology; Fundamental and applied biological sciences. Psychology; HL-60 Cells - enzymology; Humans; Male; Membrane Glycoproteins; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - enzymology; NAD+ Nucleosidase - metabolism; Rats; Rats, Sprague-Dawley; Retinoids - pharmacology; Tissue Distribution; Tretinoin - pharmacology; Triiodothyronine - pharmacology; Up-Regulation; Vertebrates: cardiovascular system; Zinc - pharmacology
• ISSN: 0009-7330; 1524-4571
• DOI: 10.1161/01.RES.86.11.1153

We investigated whether ADP-ribosyl cyclase (ADPR-cyclase) in rat vascular smooth muscle cells (VSMCs) has enzymatic properties that differ from the well-characterized CD38-antigen ADPR-cyclase, expressed in HL-60 cells. ADPR-cyclase from VSMCs, but not CD38 ADPR-cyclase from HL-60 cells, was inhibited by gangliosides (10 μmol/L) GT1B, GD1, and GM3. Preincubation of membranes from CD38 HL-60 cells, but not from VSMCs, with anti-CD38 antibodies increased ADPR-cyclase activity; CD38 antigen was detected both in VSMCs and in HL-60 cells. ADPR-cyclase in VSMC membranes was more sensitive than CD38 HL-60 ADPR-cyclase to inactivation by N- endoglycosidase F and to thermal inactivation at 45°C. The specific activity of ADPR-cyclase in membranes from VSMCs was >20-fold higher than in membranes from CD38 HL-60 cells. Most importantly, VSMC ADPR-cyclase was inhibited by Zn and Cu ions; the inhibition by Zn was dose dependent, noncompetitive, and reversible by EDTA. In contrast, Zn stimulated the activity of CD38 HL-60 ADPR-cyclase and other known types of ADPR-cyclases. Retinoids act either via the nuclear receptor retinoic acid receptor or retinoid X receptor, including all-trans retinoic acid (atRA), and panagonist 9-cis-retinoic acid–upregulated VSMC ADPR-cyclase; the stimulatory effect of atRA was blocked by actinomycin D and cycloheximide. 1,25(OH)2–Vitamin D3 (calciferol) stimulated VSMC ADPR-cyclase dose dependently at subnanomolar concentrations (ED50≅56 pmol/l). Oral administration of atRA to rats resulted in an increase of ADPR-cyclase activity in aorta (≅+60%) and, to a lesser degree, in myocardium of left ventricle (+18%), but atRA had no effect on ADPR-cyclases in lungs, spleen, intestinal smooth muscle, skeletal muscle, liver, or testis. Administration of 3,5,3′-triiodothyronine (T3) to rats resulted in an increase of ADPR-cyclase activity in aorta (≅+89%), but not in liver or brain. We conclude the following(1) ADPR-cyclase in VSMCs has enzymatic properties distinct from “classic” CD38 ADPR-cyclase, especially sensitivity to inhibition by Zn and Cu; (2) ADPR-cyclase in VSMCs is upregulated by various retinoids, calcitriol, and T3 in vitro; and (3) administration of atRA and T3 increases ADPR-cyclase in aorta in vivo. We suggest that the cADPR signaling system plays an important role in the regulation of VSMC functions in response to steroid superfamily hormones.