A Method for the Isolation of Intima-Media Samples from Arteries

Wolinsky, Harvey; Daly, Marie M.

doi:10.3181/00379727-135-35052

Cited by 65 publications

(32 citation statements)

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“…11 Intimamedia tissue samples to be assayed for lysyl oxidase activity were individually homogenized at 0° to 4° C in 2.5 volumes (ml/g wet weight) of 4 M urea, 0.016 M potassium phosphate, pH 7.7, with a Brinkmann Polytron homogenizer at high speed for 90 seconds. The homogenates were centrifuged at 1Q.000 X g for 20 minutes at 4°C, and the isolated supernatants were dialyzed vs 0.016 M potassium phosphate, pH 7.7.…”

Section: Methodsmentioning

confidence: 99%

Changes in aortic lysyl oxidase activity in diet-induced atherosclerosis in the rabbit.

Kagan¹,

Raghavan²,

Hollander³

1981

Arteriosclerosis

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This study assessed the response of lysyl oxidase, the enzyme that initiates covalent crosslinking in elastin and collagen, by studying the aortic tissue of rabbits after arteriosclerosis had been induced by diet. Rabbits in the experimental group were fed an atherogenic diet of rabbit chow supplemented with 8% peanut oil and 2% cholesterol for varying periods of time, while the control group was fed only rabbit chow. Lysyl oxidase activity was found to be distributed throughout the length of the thoracic and abdominal aortas of the normal rabbits. However, rabbits fed the atherogenic diet showed marked increases in enzyme activity in the aortic arch, a change that was initially evident after 30 days and became greatest (2.5 times that of the controls) after 90 days. Enzyme activity in the study rabbits increased only minimally in the abdominal aortic wall. Aortic prolyl hydroxylase activity measured after 60 days of feeding changed in degree and manner similar to lysyl oxidase activity. These region-specific changes in enzyme activities correlated with the distribution and severity of aortic lesions in this model of the disease. Lysyl oxidase activity increased dramatically in this model of atherosclerosis, suggesting that this extracellular enzyme activity may prove to be a vulnerable and accessible point of control of the fibrotic response in atherosclerosis. (Arteriosclerosis 1981; 1:287-291)

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Section: Methodsmentioning

confidence: 99%

Changes in aortic lysyl oxidase activity in diet-induced atherosclerosis in the rabbit.

Kagan¹,

Raghavan²,

Hollander³

1981

Arteriosclerosis

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“…Six animals from each group were used; thoracic aortic segments exactly delimited by the left 1 Testosterone cypionate, 100 mg/ml, Upjohn Company, Kalamazoo, Mich. subclavian and celiac arteries were removed and the intima and media isolated (8). After dehydration in ethanol-ether for 2 hr and ether for 1 hr as previously described (4), each aortic segment was brought to dry weight after 72 hr at 55-60°C in a vacuum oven.…”

Section: Methodsmentioning

confidence: 99%

Effects of Androgen Treatment on the Male Rat Aorta

Wolinsky¹

1972

J. Clin. Invest.

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A B S T R A C T Androgen was given to male rats to determine if it exerted effects on the aortic wvall distinct from those of estrogen deficit. Although a general anabolic effect was avoided, significant vascular effects were observed. The amounts of mural fibrous proteins, elastin and collagen, were significantly increased in treated animals; noncollagenous, alkali-soluble protein, thought to reflect the cellular component, was unchanged with treatment. These effects were not detectable on microscopic examination and measurement of the vessel wall despite attempts to duplicate closely in vivo wall dimensions before study. These findings of distinct and marked effects of androgen on vascular metabolism extend the growing evidence for an important role of sex hormones in vessel wall structure and function. INTRODUCTIONRecent studies have shown that female sex hormones exert profound effects on vessel wall morphology and metabolism. Estrogens administered to gonadectomized female rats (1, 2) and guinea pigs (3) markedlv reduce the net amount of collagen in the aortic wall and skin. Labeled precursor studies of vascular collagen in females suggest that the mechanism responsible is a combination of accelerated turnover of a relatively small pool of soluble precursors of collagen and interference

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“…Intima-media strips of the segment were quickly separated from the adventitia 13 and placed in the sucrose-EDTA solution at 4°C. Strips of segments from four rats were pooled and suspended in a final volume of 6 ml sucrose-EDTA to make each experimental sample.…”

Section: Biochemical Studiesmentioning

confidence: 99%

Hydrolase activities in the rat aorta. I. Effects of diabetes mellitus and insulin treatment.

Wolinsky¹,

Goldfischer²,

Capron³

et al. 1978

Circulation Research

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SUMMARY Vascular disease in diabetics could arise in part from altered vessel wall catebolism. Specific activities of hydrolases in aortic smooth muscle cells from rats with streptozotocin-induced diabetes were measured. Enzymes included: neutral a-glucosidase, a-mannosidase, and lysosomal /V-acetyl /3-glucosaminidase, /3-galactosidase, cathepsin C, acid a-glucosidase, and acid cholesteryl esterase. After 4 , 8 , and 11 weeks of diabetes, activities of all enzymes studied were decreased significantly in diabetic vessels, decreases ranging from 15% for cathepsin C to 62% for amannosidase. After 3 weeks of diabetes, insulin treatment for 1 week restored enzyme levels to normal. After 7 weeks of diabetes, 1 week of insulin treatment did not restore enzyme levels fully to normal (acid cholesteryl esterase was unchanged); 4 weeks of insulin did. Acid phosphatase and JV-acetyl /3-glucosaminidase activities were reduced markedly in histochemical studies of diabetic aortas at all time periods and were restored by insulin treatment. AUoxan-induced diabetes gave results similar to those with streptozotocin. Significant decreases of aortic hydrolase activities, including those of lysosomes, occur in experimental diabetes mellitus and could contribute to accumulation of substrates in vascular smooth muscle cells.

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A Method for the Isolation of Intima-Media Samples from Arteries

Cited by 65 publications

References 0 publications

Changes in aortic lysyl oxidase activity in diet-induced atherosclerosis in the rabbit.

Changes in aortic lysyl oxidase activity in diet-induced atherosclerosis in the rabbit.

Effects of Androgen Treatment on the Male Rat Aorta

Hydrolase activities in the rat aorta. I. Effects of diabetes mellitus and insulin treatment.

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