1,4-Dihydropyridine calcium channel blockers inhibit plasma and LDL oxidation and formation of oxidation-specific epitopes in the arterial wall and prolong survival in stroke-prone spontaneously hypertensive rats.
OBJECTIVE
Calcium-channel blockers (CCBs) reduce systolic blood pressure and stroke-related mortality in stroke-prone spontaneously hypertensive rats (SPSHR). Brain ischemia is associated with loss of intracellular antioxidants. Increased formation of oxygen radicals and oxidation of LDL may enhance arterial vasoconstriction by various mechanisms. CCBs that also exert antioxidative properties in vitro may therefore be particularly useful. To investigate such antioxidant effects in vivo, we determined several parameters of LDL oxidation in SPSHR treated with two 1,4-dihydropyridine-type (1,4-DHP) CCBs of different lipophilic properties and compared them with antioxidant-treated and untreated controls. We also tested whether these drugs decrease the formation of oxidation-specific epitopes in arteries.


METHODS
Five groups of 9 to 14 SPSHR each (aged 8 weeks) were treated with 80 mg/kg body wt per day nifedipine, 1 mg or 0.3 mg/kg body wt per day lacidipine, vitamin E (100 IU/d), or carrier for 5 weeks. A group of Wistar-Kyoto rats was used as normotensive control. Plasma samples were taken, and LDL was isolated by ultracentrifugation. Then LDL was exposed to oxygen radicals generated by xanthine/xanthine oxidase reaction (2 mmol/L xanthine+100 mU/mL xanthine oxidase), and several parameters of oxidation were determined. The presence of native apolipoprotein B and oxidation-specific epitopes in the carotid and middle cerebral arteries was determined immunocytochemically.


RESULTS
1,4-DHP CCBs completely prevented mortality. Normotensive Wistar-Kyoto rats showed less oxidation than control SPSHR. Plasma lipoperoxide levels were 0.87+/-0.27 micromol/L in control SPSHR, 0.69+/-0.19 and 0.63+/-0.20 micromol/L in the groups treated with 0.3 and 1 mg lacidipine, respectively, and 0.68+/-0.23 micromol/L in nifedipine-treated animals (P<0.05 versus control SPSHR for all values). Both CCBs significantly decreased formation of conjugated dienes and prolonged the lag time in LDL exposed to oxygen radicals. Similarly, lipoperoxides and malondialdehyde were significantly reduced (P<0.05). Reduced relative electrophoretic mobility and increased trinitrobenzenesulfonic acid reactivity of LDL from treated rats (P<0.01) also indicated that fewer lysine residues of apolipoprotein B were oxidatively modified in the presence of 1,4-DHP CCBs. Finally, these drugs reduced the intimal presence of apolipoprotein B and oxidized LDL (oxidation-specific epitopes) in carotid and middle cerebral arteries.


CONCLUSIONS
In the SPSHR model, 1,4-DHP CCBs reduce plasma and LDL oxidation and formation of oxidation-specific epitopes and prolong survival independently of blood pressure modifications. Our results support the concept that the in vivo protective effect of these drugs on cerebral ischemia and stroke may in part result from inhibition of oxidative processes.
Napoli
C
C
Department of Medicine and Endocrinology, University of California-San Diego, La Jolla, USA. cnapoli@ucsd.edu
Salomone
S
S
Godfraind
T
T
Palinski
W
W
Capuzzi
D M
DM
Palumbo
G
G
D'Armiento
F P
FP
Donzelli
R
R
de Nigris
F
F
Capizzi
R L
RL
Mancini
M
M
Gonnella
J S
JS
Bianchi
A
A
D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D001055:Apolipoproteins B / Q000378:metabolism; D001158:Arteries / Q000187:drug effects / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology*; D002561:Cerebrovascular Disorders / Q000235:genetics* / Q000401:mortality; D004095:Dihydropyridines / Q000494:pharmacology*; D000939:Epitopes / Q000187:drug effects / Q000378:metabolism; D020022:Genetic Predisposition to Disease; D007150:Immunohistochemistry; D008077:Lipoproteins, LDL / Q000037:antagonists & inhibitors* / Q000097:blood; D008297:Male; D010084:Oxidation-Reduction / Q000187:drug effects; D051381:Rats; D011918:Rats, Inbred SHR / Q000235:genetics* / Q000378:metabolism; D011921:Rats, Inbred WKY; D012016:Reference Values; D014810:Vitamin E / Q000494:pharmacology
D000975:Antioxidants; D001055:Apolipoproteins B; D002121:Calcium Channel Blockers; D004095:Dihydropyridines; D000939:Epitopes; D008077:Lipoproteins, LDL; C099252:oxidized low density lipoprotein; D014810:Vitamin E; C060285:lacidipine; C038806:1,4-dihydropyridine