Beruflich Dokumente
Kultur Dokumente
A. KUMMEROW•õ
Forty-six Yorkshire swine (barrows and gilts), approximately 2 months of age, were
used. The animals were divided into four groups. Group 1 (control group) consisted of
ten animals which were fed a basal ration of corn, soybean meal and a vitamin and mineral
premix containing 387 IU of vitamin D3 and 6.3 g of calcium per kg of diet. Five
animals were sacrificed at 5 months of age and the rest after 8 months of age. Group 2 (3
months D3 supplemented group) consisted of fifteen animals (see Table 2) which were
divided into five subgroups and were fed the basal ration supplemented with 125,000 IU,
62,500 IU, 31,250 IU, 6,250 IU and 1,250 IU of vitamin D3 per kg of diet for 3 months,
and then sacrificed at 5 months of age. Group 3 (6 months D3 supplemented group)
consisted of three animals which were fed 125,000 IU vitamin D3 throughout the experi
mental period and sacrificed at 8 months of age. Group 4 (vitamin D3 withdrawal group)
consisted of eighteen animals (see Table 3). They were fed the same amounts of vitamin D3
as the five subgroups of group 2 for the same time period, and subsequently received only
the basal ration for the following 3 months. Animals in group 4 were sacrificed at 8
months of age. Blood samples were taken for all animals except for group 3 at the time of
sacrifice for determination of serum cholesterol and calcium. An average of 30 coronary
artery samples (10 samples each for the three major branches) were taken from each heart for
histological evaluation.
Mean•}s.E.M.
RESULTS
The serum cholesterol and calcium levels of the experimental swine are
summarized in Table 1. The animals of 2 or 3 highest vitamin D3 fed groups in
group 2 had slightly depressed cholesterol levels and abnormally high calcium levels.
No difference in serum cholesterol and calcium levels was observed between group
4 and the control animals.
No abnormal areas of intimal thickening or calcified lesions were demonstrable
in the control group. In group 2, calcified lesions were noted in the coronary
arteries of animals fed 125,000 IU, 62,500 IU and 31,250 IU of vitamin D3 as
shown in Table 2. These calcified lesions were located in the internal elastica and
were frequently accompanied by fibromuscular intimal thickening (Fig. 1). In the
animals fed 6,250 IU and 1,250 IU of vitamin D3, intimal thickening or calcified
lesions were not observed. In group 3, prominent calcified lesions and fibromus
cular intimal thickening were noted in the coronary arteries. The incidence of
calcified lesions in the arteries was 58/98 or 51%. No lipid deposits were present
in the coronary arteries of animals from group 2 or 3.
In group 4, coronary atherosclerosis was produced in all animals initially fed
125,000 IU, 62,500 IU and 31,250 IU of vitamin D3, but in none of the animals
fed 6,250 IU or 1,250 IU of vitamin D3. The incidence of atherosclerotic lesions
was proportional to the dosage of vitamin D3 and more frequently involved the
left anterior descending artery, as shown in Table 3. An atherosclerotic intima
was always observed above the calcified internal elastica (Fig. 2). The size of the
atheromata paralleled the extent of calcification in the internal elastica; the larger
atheromata had more prominent calcified deposits in the internal elastica than
small ones. The largest atheroma was seen in the left anterior descending artery
and had the usual criteria for full-fledged atheromata having abundant lipid,
cholesterol crystals, foam cell formation, necrosis, pools of glycosaminoglycans,
fibroelastosis and calcification. The incidence of calcified lesions in the internal
elastica of samples from group 4 (Table 3) was less than that of group 2 (Table 2),
which received the same doses of vitamin D3. No intramural hemorrhage, ulcera
tion or luminal thrombosis was observed in the arteries, and no evidence of
myocardial infarction was noted in the myocardium.
Electron microscopic observations revealed many foam cells in the athero-
12 S. Taura et al.
Fig. 1. Swine, 125,000 IU vitamin D3 supplemented group (group 2), uranyl acetate and
lead citrate stains. Fibromuscular intimal thickenning (FIT) and calcified
internal elastica (Ca) are noted. Calcification is restricted in the internal elastica.
x 3600.
Fig. 2. Swine, 125,000 IU vitamin Da withdrawal group, oil red 0 and hematoxylin . Lipid
rich atheromata (A) are located above the calcified internal elastica (Ca) and show
luminal narrowing. X 200.
Fig. 3. Swine, 62,500 IU vitamin D, withdrawal group. Electron dense deposits are
noted around the internal elastica (IE). Many foam cells (F) are seen in the intima.
x 3600.
Fig. 4. Swine, 125,000 IU vitamin D, withdrawal group (group 4). Electron dense
particles are noted in the extracellular space and in the macrophage (M). Intracellular
dense particles are accompanied with lysosomal activity (L) in the surroundings. x 9000.
14 S. Taura et al.
Fig. 5. Human, 51 years old, Von Kossa and H-E stains. Calcified deposits are noted in
the internal elastica (IE). X 200.
Fig. 6. Human, 65 years old, unstained Epon specimen. Extremely dense particles
(arrows) are noted in the endothelium and intimal cells (arrows). X 3600.
Coronary Atherosclerosis in Normolipemic Swine 15
(Fig. 5), while in severe cases calcification also involved the intima and media.
Atherosclerotic involvement tended to be severe in the calcified areas. In general,
severe atherosclerotic lesions of humans were more extreme than those in the
experimental animals. But the mild to moderate atherosclerotic lesions in man had
many morphological features which resembled those in the experimental animals.
In such cases, atherosclerotic involvement was only noted above the calcified
internal elastica. Upon electron microscopic examination, extremely dense
particles were noted around the internal elastica and in the foam cells and
endothelium (Fig. 6). Microprobe analysis showed calcium to be contained within
these dense particles.
DISCUSSION
produced by simply feeding the vitamin D3 for a short period of time and then
withdrawing it from the diet.
Acknowledgment
A part of this work was presented in the 11th International Congress of Angiology at
Prague, Czechoslovakia, July 2-8,1978. We wish to thank Drs. Ben T. Williams, Fernando
B. Toledo and Sandhya Sarwate, Mercy Hospital, Urbana, IL 61801, for the specimens of
coronary arteries. This study was supported by the NHLI Grant $15504-05, the Wallace
Genetic Foundation, the Illinois Heart Association, the National Dairy Council and the
American Egg Board.
References
1) Bajwa, G.S., Morrison, L.M. & Ershoff, B.H. (1971) Induction of aortic and coronary
athero-arteriosclerosis in rats fed a hypervitaminosis D, cholesterol containing diet.
P.S.E.B.M., 138, 975-982.
2) Bjorkerud, S. & Bondjers, G. (1973) Arterial repair and atherosclerosis after mechanical
injury. Part 5. Tissue response after induction of a large superficial transverse
injury. Atherosclerosis, 18, 235-255.
3) Bulkley, B.H. & Roberts, W.C. (1976) Atherosclerotic narrowing of left main coronary
artery-Necropsy analysis of 152 patients with fatal coronary heart disease and
varying degrees of left main narrowing. Circulation, 53, 823-827.
4) Constantinidies, P., Gutmann-Auersperg, N. & Hospes, D. (1958) Acceleration of
intimal atherosclerosis through prior medial injury. Arch. Path., 66, 359-395.
5) Duguid, J.B. (1954) Diet and coronary disease. Lancet, 266, 891-895.
6) Eisenstein, R. & Zerulois, L. (1964) Vitamin D induced aortic calcification -An
electron microscopic study. Arch. Path., 77, 35-43.
7) Friedberg, C.K. (1968) Diseases of the heart. In: Disease of the Coronary Arteries
and Coronary Heart Disease. 3rd ed., Saunders Company, Philadelphia, pp. 643-693.
8) Hass, G.M., Trueheart, R.E. Taylor, C.B. & Stumple, M. (1958) An experimental
histologic study of hypervitaminosis D. Amer. J. Path., 34, 395-422.
9) Kent, S.P., Vawter, G.F. Dowben, R.M. & Benson, R.E. (1958) Hypervitaminosis
D in monkeys - A clinical and pathological study. Amer. J. Path., 34, 37-59.
10) Lansing, A.I., Blumenthal, H.T. & Gray, S.H. (1948) Aging and calcification of human
coronary artery. J. Gerontol., 3, 87-97.
11) Lee, W.M. & Lee, K.T. (1975) Advanced coronary atherosclerosis in swine produced
by combination of balloon-catheter injury and cholesterol feeding. Exp. mol. Path.,
23, 491-499.
12) Moore, S. (1973) Thromboatherosclerosis in normolipemic rabbits. A result of
continued endothelial damage. Lab. Invest., 29, 478-487.
13) Shenk, E.A., Penn, I. & Schwartz, S. (1965) Experimental atherosclerosis in the dog.
Arch. Path., 80, 102-109.
14) Woolf, N., Braldey, J.W.P., Crawford, T. & Carstairs, K.C. (1973) Experimental
mural thrombi in the pig aorta. The early natural history. Brit . J. exp. Path., 49,
257-264.