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Cocaine-induced cytopathy and apoptosis of vascular smooth muscle cells from spontaneous hypertensive rats
AJH–April 2001–VOL. 14, NO. 4, PART 2
POSTERS: Vascular Injury/Inflammation
259A
warrant further study as candidate genes altered during vascular remodeling in response to blood pressure changes. This approach has the advantage of allowing identification of genes not previously having a strong association with vascular remodeling and hypertension.
indicate that calcification of aortic arch is a predictor for ASO in elderly subjects.
Key Words: CDNA Array, Genes Smooth Muscle Cell, Expression Profile
P-686 COCAINE-INDUCED CYTOPATHY AND APOPTOSIS OF VASCULAR SMOOTH MUSCLE CELLS FROM SPONTANEOUS HYPERTENSIVE RATS
P-685 AORTIC CALCIFICATION AS A MARKER FOR ARTERIOSCLEROSIS OBLITERANS OF LOWER EXTREMITIES IN THE ELDERLY Masashi Okuro, Yoshichika Baba, Miyuki Onishi, Kohya Okaishi, Shoroku Sakurai, Noriko Fujiwara, Jin Yang, Ryuichi Morishita, Shigeto Morimoto, Toshio Ogihara. 1Geriatric Medicine, Osaka University Medical School, Suita, Osaka, Japan [Aim] Arteriosclerosis obliterans (ASO) is one of the major health problems in the elderly, since this disease causes decreased ADL and QOL. On the other hand, clinical significance of calcification of aortic arch in this disease has not been evaluated in the elderly. In this study, we examined whether there is association of aortic calcification and ASO in the elderly after adjustment of known risk factors including hypertension. [Method] The subjects were elderly patients [mean⫾SD age: 83⫾8 (SD) years] with (n⫽33, ankle-brachial pressure index: ABI⬍0.2) or without (n⫽82, ABI⬎0.9) ASO. We classified calcification of aortic arch on the chest X-ray into 4 groups as no (grade 0), speckled (grade 1), linear (grade 2) and circular (grade 3) calcification. [Result] The distribution of grades of the calcification (0 to 3) were 9%, 24%, 46%, 21% in ASO patients, and 22%, 42%, 31%, 6% in control subjects, respectively (ANOVA p⫽0.013). Hypertension itself was associated with the occurrence of ASO by univariate analysis. Logistic regression analysis using diabetes mellitus, hypertension, hypercholesterolemia and high serum Lp(a) as confounding factors revealed that the aortic calcification is the highest risk factor among these confounding factors. [Discussion] ASO is known to occur in iliac and femoral arteries which comparatively locate near to the aorta. In consequence, our results
Key Words: arteriosclerosis obliterans, aortic calcification, hypertension
K. Li, J. H. Wang, I. Hale, H. J. Ward. 1Internal Medicine, UCLA and C. Drew Schools of Medicine, Los Angeles, CA, United States There is increasing clinical evidence to document cocaine abuse as a risk factor for progressive cardio-renal disease, especially in hypertensive individuals. The present study was designed to define the vasculotoxic and nephrotoxic effects of cocaine at the cellular level. The smooth muscle cells were isolated from the aorta of 8-week old spontaneous hypertensive rats. Cultured vascular smooth muscle cells at passages between 8 and 10 were exposed to cocaine concentrations ranging from 0.1 M to 1 mM for 24 and 72 hours. At a cocaine concentration of 0.1 mM, phase contrast microscopy revealed cytoplasmic vacuolization after 24-hour incubation. The highest cocaine concentration of 1 mM was associated with the development of early apoptotic changes of cell retraction and chromatin condensation. Chromatin condensation was also detected as bright fluorescent staining of DNA by Hoechst 35228, distributed as a granular pattern in the nucleus or as apoptotic bodies. Prolonged (72 hours) cocaine exposure was accompanied by nuclear shrinkage and cell detachment. The apoptotic effect of cocaine on vascular smooth muscle cells was further confirmed by DNA fragmentation using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. After 24-hour incubation with cocaine, positive TUNEL stained cells were observed at the concentration of 1 mM. In vascular smooth muscle cells exposed to cocaine for 72 hours, TUNEL assay showed a dose-dependent apoptotic effect at concentrations of 0.03 mM and higher. These direct cytopathic changes and the pro-apoptotic effects of cocaine support the clinical observations of increased cardiorenal morbidity in cocaine abuser. Key Words: Cocaine, Apoptosis, Vascular smooth muscle cell
POSTERS: Vascular Injury/Inflammation
259A
warrant further study as candidate genes altered during vascular remodeling in response to blood pressure changes. This approach has the advantage of allowing identification of genes not previously having a strong association with vascular remodeling and hypertension.
indicate that calcification of aortic arch is a predictor for ASO in elderly subjects.
Key Words: CDNA Array, Genes Smooth Muscle Cell, Expression Profile
P-686 COCAINE-INDUCED CYTOPATHY AND APOPTOSIS OF VASCULAR SMOOTH MUSCLE CELLS FROM SPONTANEOUS HYPERTENSIVE RATS
P-685 AORTIC CALCIFICATION AS A MARKER FOR ARTERIOSCLEROSIS OBLITERANS OF LOWER EXTREMITIES IN THE ELDERLY Masashi Okuro, Yoshichika Baba, Miyuki Onishi, Kohya Okaishi, Shoroku Sakurai, Noriko Fujiwara, Jin Yang, Ryuichi Morishita, Shigeto Morimoto, Toshio Ogihara. 1Geriatric Medicine, Osaka University Medical School, Suita, Osaka, Japan [Aim] Arteriosclerosis obliterans (ASO) is one of the major health problems in the elderly, since this disease causes decreased ADL and QOL. On the other hand, clinical significance of calcification of aortic arch in this disease has not been evaluated in the elderly. In this study, we examined whether there is association of aortic calcification and ASO in the elderly after adjustment of known risk factors including hypertension. [Method] The subjects were elderly patients [mean⫾SD age: 83⫾8 (SD) years] with (n⫽33, ankle-brachial pressure index: ABI⬍0.2) or without (n⫽82, ABI⬎0.9) ASO. We classified calcification of aortic arch on the chest X-ray into 4 groups as no (grade 0), speckled (grade 1), linear (grade 2) and circular (grade 3) calcification. [Result] The distribution of grades of the calcification (0 to 3) were 9%, 24%, 46%, 21% in ASO patients, and 22%, 42%, 31%, 6% in control subjects, respectively (ANOVA p⫽0.013). Hypertension itself was associated with the occurrence of ASO by univariate analysis. Logistic regression analysis using diabetes mellitus, hypertension, hypercholesterolemia and high serum Lp(a) as confounding factors revealed that the aortic calcification is the highest risk factor among these confounding factors. [Discussion] ASO is known to occur in iliac and femoral arteries which comparatively locate near to the aorta. In consequence, our results
Key Words: arteriosclerosis obliterans, aortic calcification, hypertension
K. Li, J. H. Wang, I. Hale, H. J. Ward. 1Internal Medicine, UCLA and C. Drew Schools of Medicine, Los Angeles, CA, United States There is increasing clinical evidence to document cocaine abuse as a risk factor for progressive cardio-renal disease, especially in hypertensive individuals. The present study was designed to define the vasculotoxic and nephrotoxic effects of cocaine at the cellular level. The smooth muscle cells were isolated from the aorta of 8-week old spontaneous hypertensive rats. Cultured vascular smooth muscle cells at passages between 8 and 10 were exposed to cocaine concentrations ranging from 0.1 M to 1 mM for 24 and 72 hours. At a cocaine concentration of 0.1 mM, phase contrast microscopy revealed cytoplasmic vacuolization after 24-hour incubation. The highest cocaine concentration of 1 mM was associated with the development of early apoptotic changes of cell retraction and chromatin condensation. Chromatin condensation was also detected as bright fluorescent staining of DNA by Hoechst 35228, distributed as a granular pattern in the nucleus or as apoptotic bodies. Prolonged (72 hours) cocaine exposure was accompanied by nuclear shrinkage and cell detachment. The apoptotic effect of cocaine on vascular smooth muscle cells was further confirmed by DNA fragmentation using the terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL) assay. After 24-hour incubation with cocaine, positive TUNEL stained cells were observed at the concentration of 1 mM. In vascular smooth muscle cells exposed to cocaine for 72 hours, TUNEL assay showed a dose-dependent apoptotic effect at concentrations of 0.03 mM and higher. These direct cytopathic changes and the pro-apoptotic effects of cocaine support the clinical observations of increased cardiorenal morbidity in cocaine abuser. Key Words: Cocaine, Apoptosis, Vascular smooth muscle cell