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Vitamin E and lipoproteins in hyperlipoproteinemia
Atherosclerosis, 53 (1984) 327-330 Elsevier Scientific Publishers Ireland,
327 Ltd.
ATH 03570
Preliminary Note
Vitamin E and Lipoproteins in Hyperlipoproteinemia Daniel Lambert and Jacques Mourot Inserm U. 59, Nutrition et Dibktique,
Universitk de Nancy, 38 Rue Lionnois, 54000 Nancy (France)
(Received 26 March, 1984) (Revised, received 6 August, 1984) (Accepted 10 August, 1984)
suuunluy The composition of vitamin E in serum and lipoproteins was determined in type I, IIa, IIb, IV and IV hyperlipoproteinemia and in normal subjects. Vitamin E was not specifically associated with any one of the lipoproteins but increased vitamin E levels were observed in VLDL when triacylglycerols level was increased (types IIb and IV); the same observations were noted in LDL when cholesterol level was increased (type IIa). Key words: Alpha-tocopherol
- Hyperlipoproteinemia - Vitamin
Introduction
Alpha-tocopherol plays a role in the prevention of peroxidative damage to cells and subcellular elements [1,2]. It is known to prevent platelet aggregation [3,4] and may be a beneficial effector in the prevention of cardiovascular diseases [5,6]. Alpha-tocopherol is associated with LP [7]. However, controversial results describe the higher level of vit. E present either in LDL [8,9] or HDL [7,10]. The aim of this study was to determine: (1) Whether the vit. E level in normal This investigation was supported by a grant from “Groupe Lipide et Nutrition”. Address all correspondence to: Dr. D. Lambert, Inserm U 59, 38 rue Lionnois, WI00 Nancy, France. Abbreviations: HDL = high density lipoproteins; HLP = hyperl@oproteinemia; LDL = low density lipoproteins; LP = lipoproteins; TC = total cholesterol; TG = triacylglycerols; Vit. E = vitamin E; VLDL - very low density lipoproteins.
0021-9150/84/$03.00
0 1984 Elsevier Scientific
Publishers
Ireland,
Ltd.
328
subjects is higher in LDL or HDL; (2) Whether the pathologically elevated LP differ from normal LP in their content of vit. E. Subjects and Methods
The study was carried out on 11 normal and 26 hyperlipoproteinemic male subjects without drug or dietetic treatment. Type IIa: n = 8; IIb: n = 7; IV: n = 8; I: n = 2; V: n = 1. The HLP was confirmed by analysis of TC and TG levels in LP according to the WHO classification [ll]. Following a 12-h overnight fast, TC (mM), TG (mM) and vit. E (PM) assays were performed in serum and LP isolated by ultracentrifugation [12]. CT [13] and TG [14] were measured using enzymatic techniques. The vit. E level was measured using high-performance liquid chromatography [15]. Results Serum and lipoproteins and vitamin E in normal subjects
Highest values for vit. E were found in LDL (13.27 * 0.71 pmoles/l) (Table 1). Normolipoproteinemic subjects revealed a significant correlation between TC and vit. E in HDL (r = 0.83, P -C0.001) and between TG and vit. E in VLDL (r = 0.92, P -C 0.001) (Fig. 1). Serum vitamin E and hyperlipoproteinemia
There was a significant increase in the vit. E level in type IIa HLP (P < 0.01) and type IIb (P < 0.05) compared to normal subjects. There was no significant variation in vit. E level in type IV HLP, although an increase was observed. In types I and V HLP, increases in vit. E levels were observed, although no statistical analysis was possible. Lipoprotein
vitamin E and HLP
Type IIa showed a significant increase in vit. E levels in LDL (P < 0.001) and in VLDL (P c 0.05) (Table 1).
TABLE 1 VALUES FOR VIT. E (pM) IN SERUM AND LP, FROM NORMAL
(N) AND HLP SUBJECTS
Means are f SE, t-test against N. Serum N
n=ll
28.31* 2.87
IIa IIb IV I V
n= 8 n= 7 n=8 n=2 n=l
46.06k5.33 ** 41.66k4.44 * 35.64k4.19 40.20* 15.00 51.27
Chylo 0.00 0.00 1.62*0.23 *** 2.34f0.52 l ** 39.50* 14.50 29.27
* P < 0.05; ** P c 0.01; *** P < 0.001
VLDL
LDL
HDL
2.72 f 0.30 5.59*1.14 * 12.40* 1.40 *** 15.23 i2.53 *** 13.50 f 1.20 24.19
13.27 f 0.71 28.23k2.84 *** 21.04* 1.76 *** 12.12& 1.76 8.70* 0.50 6.71
10.47 f 0.94 11.15f1.37 8.54f 1.32 9.15 f 1.49 9.75 f 2.70 5.27
329 CHYLOMICRONS 41
.
30
40
50
0
5
10
15
20
25
Fig. 1. Vitamin E, cholesterol and triacylglycerols in LP. Correlations as a function of HLP. (0) normal type I HLP; (A) type IIa HLP; (A) type IIb HLP; (m) type IV HLP; (0) type V HLP.
subject: (*)
In type IIb HLP, chylomicrons had high values of vit. E (P < 0.001). A very significant increase (P < 0.001) was observed in VLDL-vit. E and LDL-vit. E. In type IV HLP, chylomicrons had high values of vit. E (P -E0.001). There was also a significant increase in VLDl-vit. E (P < 0.001). In type I and V HLP, we observed a very high level of vit. E mainly associated with chylomicrons and VLDL, respectively. Significant correlations were noted between TC and Vit. E in LDL (I = 0.69, P < 0.05) in type IIa, and in LDL in type IIb (r = 0.70, P -c0.05). A TG-vit. E in type IIa HLP and correlation was only noted within VLDL (r = 0.70, P -c0.05) within chylomicrons (r = 0.84, P < 0.01) and VLDL (r = 0.88, P < 0.001) in type IV HLP. Discussion
The present study shows that the greatest concentration of vit. E is found in LDL, thus contrary to Takahashi et al. [7], who showed the highest concentration occurring in HDL, especially HDL, (28%). The results obtained by Lewis et al [lo] also vary from our own. However, in these 2 papers, the percentage of HDL was measured indirectly. Examination of the distribution of vit. E in the different types of HLP shows that there is an increase in the vit. E level of the specific LP modified by the disease. In other words, when the TG level increases (type I, IIb, IV and V HLP), the vit. E level increases in VLDL and vit. E is seen in association with TG. When TC levels are increased (type IIa and IIb HLP), the vit. E level increases in LDL. We can
330
therefore deduce that when the level of lipids increases, whatever irrespective of the TC or TG levels, the vit. E level increases in the corresponding lipid transport protein. Correlations between vit. E and lipids appear in the LP affected by HLP. It is interesting to note that the vit. E/TC correlation only exists in HDL and not in LDL in normal subjects, which agrees with the hypothesis of a beneficial role of vit. E in atherosclerosis [3,16,17]. However, as little is known concerning transport of vit. E to various tissues [7], further interpretations need to be considered carefully. Serum vit. E generally reflects concentrations in tissues. However, this could be wrong in HLP, where an excess of vit. E lipid carrier could be responsible for the deficiency at tissue level [18]. References 1 Fonck, K. and Konings, A.W., The effect of vitamin E on cellular survival after X irradiation of lymphoma cells, Brit. J. Radiol., 51 (1978) 832. 2 Nakamura, T. and Masugi, F., Transfer of alpha-tocopherol from plasma to erythrocytes in vitamin E-deficient rats, Int. J. Vit. Nutr. Res., 49 (1979) 364. 3 Machlin, L.J., Filipski, R., Willis, A.L., Kuhn, D.C. and Brin, M., Influence of vitamin E on platelet aggregation and thrombocythemia in the rat, Proc. Sot. Exp. Biol. Med., 149 (1975) 275. 4 Steiner, M., Vitamin E changes the membrane fluidity of human platelets, Biochim. Biophys. Acta, 640 (1981) 100. 5 Haeger, K., The treatment of peripheral occlusive arterial disease with a-tocopherol as compared with vasodilator agents and antiprothrombin (dicumarol), Vast. Dis., 5 (1968) 199. 6 Haeger, K., Long-time treatment of intermittent claudication with vitamin E, Amer. J. Clin. Nutr., 27 (1974) 1179. 7 Takahashi Y., Urono K. and Kimura S., Vitamin E binding proteins in human serum, J. Nutr. Sci. Vit., 23 (1977) 201. 8 McCormick, E.C., Cornwell, D.G. and Brown, J.B., Studies on the distribution of tocopherol in human serum lipoproteins, J. Lipid. Res., 1 (1960) 221. 9 Kayden, H.J. and Bjomson, L., The dynamics of vitamin E transport in the human erythrocyte, Ann. N.Y. Acad. Sci., 203 (1972) 127. 10 Lewis, L.A., Quaife, M.L. and Page, I.H., Lipoproteins of serum, carriers of tocopherol, Amer. J. Physiol., 178 (1954) 222. 11 Beaumont, J.L., Carlson, L.A., Cooper, G.R., Fejfar, Z., Fredrickson, D.S. and- Strasser, N.T., Classification of hyperlipidemias and hyperlipoproteinemias, Bull. WHO, 43 (1970) 891. 12 Havel, R.J., Eder, H.A. and Bragdon, J.H., The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum, J. Clin. Invest., 34 (1955) 1345. 13 Klose, S., Hagen, A. and Greif, H., Mtthode de dosage colorimetrique du cholesterol par voie entierement enzymatique adapt&e B tous les types d’auto-analyseurs. In: G. Siest (Ed.), Organisation des Laboratoires - Biologie Prospective, IIIe Colloque de Pont-a-Mousson, L’Expansion Scientifique Francaise, Paris, 1975, p. 505. 14 Wahlefeld, A.W., Triglycerides determination after enzymatic hydrolysis. In: H.U. Bergmeyer (Ed.), Methods of Enzymatic Analysis, Vol. 4, 2nd edition, Verlag Chemie/Academic Press, Stuttgart, New York, 1974, p. 1831. 15 Heng, E.L., Klein, D. and Debry, G., Analytical method for the determination of plasma vitamin E by high performance liquid chromatography, Int. J. Vit. Nutr. Res., 53(3) (1983) 280. 16 Butturini. U., Vitamins E and A in vascular diseases, Acta Vit. Enzymol., 4 (1982) 15. 17 Barboriak, J.J., Ghaitit, E.P., Shetty, K.R. and Kalbfleish, J.H., Vitamin E supplements and plasma high density lipoprotein cholesterol, Amer. J. Clin. Path., 77 (1982) 371. 18 Ostrowski, J., Hypertocopherolaemia in a patient with impaired tocopherol (vitamin E) absorption, Polski Tyg. Lek., 36(2) (1981) 65.
327 Ltd.
ATH 03570
Preliminary Note
Vitamin E and Lipoproteins in Hyperlipoproteinemia Daniel Lambert and Jacques Mourot Inserm U. 59, Nutrition et Dibktique,
Universitk de Nancy, 38 Rue Lionnois, 54000 Nancy (France)
(Received 26 March, 1984) (Revised, received 6 August, 1984) (Accepted 10 August, 1984)
suuunluy The composition of vitamin E in serum and lipoproteins was determined in type I, IIa, IIb, IV and IV hyperlipoproteinemia and in normal subjects. Vitamin E was not specifically associated with any one of the lipoproteins but increased vitamin E levels were observed in VLDL when triacylglycerols level was increased (types IIb and IV); the same observations were noted in LDL when cholesterol level was increased (type IIa). Key words: Alpha-tocopherol
- Hyperlipoproteinemia - Vitamin
Introduction
Alpha-tocopherol plays a role in the prevention of peroxidative damage to cells and subcellular elements [1,2]. It is known to prevent platelet aggregation [3,4] and may be a beneficial effector in the prevention of cardiovascular diseases [5,6]. Alpha-tocopherol is associated with LP [7]. However, controversial results describe the higher level of vit. E present either in LDL [8,9] or HDL [7,10]. The aim of this study was to determine: (1) Whether the vit. E level in normal This investigation was supported by a grant from “Groupe Lipide et Nutrition”. Address all correspondence to: Dr. D. Lambert, Inserm U 59, 38 rue Lionnois, WI00 Nancy, France. Abbreviations: HDL = high density lipoproteins; HLP = hyperl@oproteinemia; LDL = low density lipoproteins; LP = lipoproteins; TC = total cholesterol; TG = triacylglycerols; Vit. E = vitamin E; VLDL - very low density lipoproteins.
0021-9150/84/$03.00
0 1984 Elsevier Scientific
Publishers
Ireland,
Ltd.
328
subjects is higher in LDL or HDL; (2) Whether the pathologically elevated LP differ from normal LP in their content of vit. E. Subjects and Methods
The study was carried out on 11 normal and 26 hyperlipoproteinemic male subjects without drug or dietetic treatment. Type IIa: n = 8; IIb: n = 7; IV: n = 8; I: n = 2; V: n = 1. The HLP was confirmed by analysis of TC and TG levels in LP according to the WHO classification [ll]. Following a 12-h overnight fast, TC (mM), TG (mM) and vit. E (PM) assays were performed in serum and LP isolated by ultracentrifugation [12]. CT [13] and TG [14] were measured using enzymatic techniques. The vit. E level was measured using high-performance liquid chromatography [15]. Results Serum and lipoproteins and vitamin E in normal subjects
Highest values for vit. E were found in LDL (13.27 * 0.71 pmoles/l) (Table 1). Normolipoproteinemic subjects revealed a significant correlation between TC and vit. E in HDL (r = 0.83, P -C0.001) and between TG and vit. E in VLDL (r = 0.92, P -C 0.001) (Fig. 1). Serum vitamin E and hyperlipoproteinemia
There was a significant increase in the vit. E level in type IIa HLP (P < 0.01) and type IIb (P < 0.05) compared to normal subjects. There was no significant variation in vit. E level in type IV HLP, although an increase was observed. In types I and V HLP, increases in vit. E levels were observed, although no statistical analysis was possible. Lipoprotein
vitamin E and HLP
Type IIa showed a significant increase in vit. E levels in LDL (P < 0.001) and in VLDL (P c 0.05) (Table 1).
TABLE 1 VALUES FOR VIT. E (pM) IN SERUM AND LP, FROM NORMAL
(N) AND HLP SUBJECTS
Means are f SE, t-test against N. Serum N
n=ll
28.31* 2.87
IIa IIb IV I V
n= 8 n= 7 n=8 n=2 n=l
46.06k5.33 ** 41.66k4.44 * 35.64k4.19 40.20* 15.00 51.27
Chylo 0.00 0.00 1.62*0.23 *** 2.34f0.52 l ** 39.50* 14.50 29.27
* P < 0.05; ** P c 0.01; *** P < 0.001
VLDL
LDL
HDL
2.72 f 0.30 5.59*1.14 * 12.40* 1.40 *** 15.23 i2.53 *** 13.50 f 1.20 24.19
13.27 f 0.71 28.23k2.84 *** 21.04* 1.76 *** 12.12& 1.76 8.70* 0.50 6.71
10.47 f 0.94 11.15f1.37 8.54f 1.32 9.15 f 1.49 9.75 f 2.70 5.27
329 CHYLOMICRONS 41
.
30
40
50
0
5
10
15
20
25
Fig. 1. Vitamin E, cholesterol and triacylglycerols in LP. Correlations as a function of HLP. (0) normal type I HLP; (A) type IIa HLP; (A) type IIb HLP; (m) type IV HLP; (0) type V HLP.
subject: (*)
In type IIb HLP, chylomicrons had high values of vit. E (P < 0.001). A very significant increase (P < 0.001) was observed in VLDL-vit. E and LDL-vit. E. In type IV HLP, chylomicrons had high values of vit. E (P -E0.001). There was also a significant increase in VLDl-vit. E (P < 0.001). In type I and V HLP, we observed a very high level of vit. E mainly associated with chylomicrons and VLDL, respectively. Significant correlations were noted between TC and Vit. E in LDL (I = 0.69, P < 0.05) in type IIa, and in LDL in type IIb (r = 0.70, P -c0.05). A TG-vit. E in type IIa HLP and correlation was only noted within VLDL (r = 0.70, P -c0.05) within chylomicrons (r = 0.84, P < 0.01) and VLDL (r = 0.88, P < 0.001) in type IV HLP. Discussion
The present study shows that the greatest concentration of vit. E is found in LDL, thus contrary to Takahashi et al. [7], who showed the highest concentration occurring in HDL, especially HDL, (28%). The results obtained by Lewis et al [lo] also vary from our own. However, in these 2 papers, the percentage of HDL was measured indirectly. Examination of the distribution of vit. E in the different types of HLP shows that there is an increase in the vit. E level of the specific LP modified by the disease. In other words, when the TG level increases (type I, IIb, IV and V HLP), the vit. E level increases in VLDL and vit. E is seen in association with TG. When TC levels are increased (type IIa and IIb HLP), the vit. E level increases in LDL. We can
330
therefore deduce that when the level of lipids increases, whatever irrespective of the TC or TG levels, the vit. E level increases in the corresponding lipid transport protein. Correlations between vit. E and lipids appear in the LP affected by HLP. It is interesting to note that the vit. E/TC correlation only exists in HDL and not in LDL in normal subjects, which agrees with the hypothesis of a beneficial role of vit. E in atherosclerosis [3,16,17]. However, as little is known concerning transport of vit. E to various tissues [7], further interpretations need to be considered carefully. Serum vit. E generally reflects concentrations in tissues. However, this could be wrong in HLP, where an excess of vit. E lipid carrier could be responsible for the deficiency at tissue level [18]. References 1 Fonck, K. and Konings, A.W., The effect of vitamin E on cellular survival after X irradiation of lymphoma cells, Brit. J. Radiol., 51 (1978) 832. 2 Nakamura, T. and Masugi, F., Transfer of alpha-tocopherol from plasma to erythrocytes in vitamin E-deficient rats, Int. J. Vit. Nutr. Res., 49 (1979) 364. 3 Machlin, L.J., Filipski, R., Willis, A.L., Kuhn, D.C. and Brin, M., Influence of vitamin E on platelet aggregation and thrombocythemia in the rat, Proc. Sot. Exp. Biol. Med., 149 (1975) 275. 4 Steiner, M., Vitamin E changes the membrane fluidity of human platelets, Biochim. Biophys. Acta, 640 (1981) 100. 5 Haeger, K., The treatment of peripheral occlusive arterial disease with a-tocopherol as compared with vasodilator agents and antiprothrombin (dicumarol), Vast. Dis., 5 (1968) 199. 6 Haeger, K., Long-time treatment of intermittent claudication with vitamin E, Amer. J. Clin. Nutr., 27 (1974) 1179. 7 Takahashi Y., Urono K. and Kimura S., Vitamin E binding proteins in human serum, J. Nutr. Sci. Vit., 23 (1977) 201. 8 McCormick, E.C., Cornwell, D.G. and Brown, J.B., Studies on the distribution of tocopherol in human serum lipoproteins, J. Lipid. Res., 1 (1960) 221. 9 Kayden, H.J. and Bjomson, L., The dynamics of vitamin E transport in the human erythrocyte, Ann. N.Y. Acad. Sci., 203 (1972) 127. 10 Lewis, L.A., Quaife, M.L. and Page, I.H., Lipoproteins of serum, carriers of tocopherol, Amer. J. Physiol., 178 (1954) 222. 11 Beaumont, J.L., Carlson, L.A., Cooper, G.R., Fejfar, Z., Fredrickson, D.S. and- Strasser, N.T., Classification of hyperlipidemias and hyperlipoproteinemias, Bull. WHO, 43 (1970) 891. 12 Havel, R.J., Eder, H.A. and Bragdon, J.H., The distribution and chemical composition of ultracentrifugally separated lipoproteins in human serum, J. Clin. Invest., 34 (1955) 1345. 13 Klose, S., Hagen, A. and Greif, H., Mtthode de dosage colorimetrique du cholesterol par voie entierement enzymatique adapt&e B tous les types d’auto-analyseurs. In: G. Siest (Ed.), Organisation des Laboratoires - Biologie Prospective, IIIe Colloque de Pont-a-Mousson, L’Expansion Scientifique Francaise, Paris, 1975, p. 505. 14 Wahlefeld, A.W., Triglycerides determination after enzymatic hydrolysis. In: H.U. Bergmeyer (Ed.), Methods of Enzymatic Analysis, Vol. 4, 2nd edition, Verlag Chemie/Academic Press, Stuttgart, New York, 1974, p. 1831. 15 Heng, E.L., Klein, D. and Debry, G., Analytical method for the determination of plasma vitamin E by high performance liquid chromatography, Int. J. Vit. Nutr. Res., 53(3) (1983) 280. 16 Butturini. U., Vitamins E and A in vascular diseases, Acta Vit. Enzymol., 4 (1982) 15. 17 Barboriak, J.J., Ghaitit, E.P., Shetty, K.R. and Kalbfleish, J.H., Vitamin E supplements and plasma high density lipoprotein cholesterol, Amer. J. Clin. Path., 77 (1982) 371. 18 Ostrowski, J., Hypertocopherolaemia in a patient with impaired tocopherol (vitamin E) absorption, Polski Tyg. Lek., 36(2) (1981) 65.