In vivo transfer of antisense oligonucleotide against urinary kininase blunts deoxycorticosterone acetate-salt hypertension in rats.
We have previously reported that the renal kallikrein-kinin system suppressed the development of deoxycorticosterone acetate (DOCA)-salt hypertension. Kinins were degraded in the kidney mainly by carboxypeptidase Y (CPY)-like kininase. Blockade of renal kinin degradation may reduce hypertension in the developmental stage. We constructed an antisense oligonucleotide against rat CPY homologue (5'-CAT-CTC-TGC-TTC-CTT-GTG-TC-3', AS) and its randomized control oligonucleotide (5'-TCC-TTC-CTG-CTT-GAG-TTC-CT-3', RC), and prepared an HVJ-liposome complex that prolongs and increases the effectiveness of the antisense oligonucleotide. Antisense oligonucleotide was transfected (25 nmole rat(-1), in terms of nucleotide) into the kidney from the renal artery. Blood pressure was measured through a catheter inserted into the abdominal aorta. Mean blood pressure (MBP) in DOCA-salt treated (for 2 weeks) Sprague Dawley strain rats was 130+/-3 mmHg (n=11), and was reduced significantly (P<0.05) more by AS transfection (122+/-4 mmHg, n=6) than by RC treatment (137+/-6 mmHg, n=5) 4 days after the transfection. This reduction in MBP was accompanied by increased urinary sodium excretion (AS, 8.4+/-1.5 mmole day(-1); RC, 4.6+/-0.5 mmole day(-1), P<0.05) and a reduction in urinary CPY-like kininase activity. Ebelactone B (5 mg kg(-1), twice a day, p.o.), an inhibitor for urinary CPY-like kininase, also reduced MBP and induced natriuresis to the same degree as AS. Lisinopril, an inhibitor for angiotensin converting enzyme (ACE) failed to reduce the elevated MBP. These results suggest that CPY-like kininase may have more contribution than ACE to degrade kinin in the kidney, and that knockdown of CPY-like kininase in the kidney may partly prevent rat DOCA-salt hypertension.
Hayashi
I
I
Department of Pharmacology, Kitasato University School of Medicine, 1-15-1 Kitasato, Sagamihara, Kanagawa 228-8555, Japan. hayashii@med.kitasato-u.ac.jp
Majima
M
M
Fujita
T
T
Okumura
T
T
Kumagai
Y
Y
Tomita
N
N
Morishita
R
R
Higaki
J
J
Ogiwara
T
T
D000818:Animals; D002268:Carboxypeptidases / Q000037:antagonists & inhibitors / Q000502:physiology*; D043402:Cathepsin A; D003900:Desoxycorticosterone; D006973:Hypertension / Q000188:drug therapy*; D007705:Kinins / Q000378:metabolism; D008297:Male; D016376:Oligonucleotides, Antisense / Q000627:therapeutic use*; D007703:Peptidyl-Dipeptidase A / Q000502:physiology; D051381:Rats; D017207:Rats, Sprague-Dawley; D012965:Sodium Chloride; D014162:Transfection
D007705:Kinins; D016376:Oligonucleotides, Antisense; D003900:Desoxycorticosterone; D012965:Sodium Chloride; D002268:Carboxypeptidases; D007703:Peptidyl-Dipeptidase A; D043402:Cathepsin A