Long chain polyunsaturated fatty acids may account for higher low-density lipoprotein oxidation susceptibility in Lithuanian compared to Swedish men.

OBJECTIVES: Mortality in coronary heart disease among middle-aged men is four times higher in Lithuania than in Sweden. Traditional risk factors cannot account for this difference. We earlier reported that low-density lipoprotein (LDL) in Lithuanian men showed a lower resistance to oxidation, measured as LDL lag time during copper oxidation, than that in Swedish men. Serum concentrations of several fat-soluble antioxidant vitamins were lower among Lithuanian men. The aim of this study was to investigate whether differences in LDL fatty acid composition could account for the difference in LDL oxidation susceptibility between men in the two countries. METHODS: This cross-sectional study included randomly selected healthy 50-year-old men from Vilnius, Lithuania (n=50) and Linköping, Sweden (n=50). Main outcome measures were fatty acids in LDL, phospholipid (PL) and cholesterol ester (CE) fractions of LDL and LDL oxidation susceptibility. RESULTS: The mean proportions of PL 20:5n3 (eicosapentaenoic acid, EPA) were higher in Vilnius men (2.09 +/- 1.05 vs. 1.53 +/- 0.58%, p= 0.004). LDL lag time was shorter in Vilnius men, mean +/- SD (75.4 +/- 13.6 vs. 89.5 +/- 13.1 mins, p<0.0001) than in Linköping men. Mean serum gamma-tocopherol was lower in Vilnius men (0.07 +/- 0.05 vs. 0.12 +/- 0.04 microg/mmol, p<0.0001) but alpha-tocopherol did not differ. In a multiple regression analysis controlled for city, high PL-EPA, low alpha-tocopherol, and high plasma triglycerides significantly contributed to a short LDL lag time, r2=0.53. CONCLUSIONS: Fat quality, i.e. poly unsaturated fatty acids, especially LDL-EPA, plasma triglycerides and antioxidative vitamins may partly account for the increased LDL oxidation susceptibility found in Vilnius men compared with Linköping men.

Adipose tissue fatty acids in men from two populations with different cardiovascular risk: the LiVicordia study.

The LiVicordia study was set up to investigate possible causes for coronary heart disease mortality in middle-aged Lithuanian men being four times higher than in Swedish men. In a previous part of this study we found lower total and low density lipoprotein (LDL) cholesterol in the Lithuanian men in spite of them having a higher fat intake than in the Swedish men. Their LDL was also more susceptible to oxidation in vitro than was that of the Swedish men. Fat quality can influence LDL oxidation. In order to obtain data on long-term fat quality intake we measured the fatty acid composition of abdominal wall adipose tissue by gas chromatography in men aged 50 years from Vilnius, Lithuania (n=50) and Linköping, Sweden (n=50). Men from Vilnius had a significantly higher percentage of adipose tissue long chain polyunsaturated fatty acids (PUFA) (20:4n6, 20:5n3, 22:5n5, 22:6n3) and lower percentage of saturated fatty acids, especially myristic acid (14:0), 3.4+/-0.7 versus 4.6+/-0.8, p<0.0001. The percentage content of adipose tissue linoleic acid (18:2n6) was 11.5+/-2.1 versus 11.0+/-1.4 (n.s.) and of linolenic acid (18:3n3) 0.7+/-0.3 versus 0.6+/-0.2 (n.s.) in men from Vilnius and Linköping, respectively. It is concluded that the adipose tissue content of essential fatty acids is similar in men from Vilnius and men from Linköping and therefore the intake is also likely to be similar. The higher contents of long chain highly unsaturated fatty acids in men from Vilnius may be of importance in the oxidation process of LDL.

Increased plasma 7 beta-hydroxycholesterol concentrations in a population with a high risk for cardiovascular disease.

The mortality in coronary heart disease among 50- to 54-year-old men is 4 times higher in Lithuania than in Sweden. It was recently suggested that traditional risk factors could not explain this mortality difference. LDL of Lithuanian men showed, however, a lower resistance to oxidation than that of Swedish men. In addition, the plasma concentration of gamma-tocopherol, lycopene, and beta-carotene were lower in Lithuanian men. In the present investigation, we determined plasma oxysterols in men from Lithuania and Sweden and found that the plasma concentration of 7 beta-hydroxycholesterol was higher in Lithuanian men, 12+/-5 versus 9+/-8 (SD) ng/mL (P=0.0011). This oxysterol is a cholesterol autoxidation product and there is no indication that it should have an enzymatic origin. Mean LDL oxidation lag time was shorter in Lithuanian men (75+/-14 versus 90+/-13 minutes, P<0.0001) and the concentration of LDL linoleic acid was lower (249+/-56 versus 292+/-54 microgram/mg of LDL protein, P<0.0001). Lipid corrected gamma-tocopherol was 0.07+/-0.02 mg/mL in Vilnius men and 0.12+/-0. 04 mg/mL (P<0.0001) in Linköping men. There was a negative correlation between the concentration of 7 beta-hydroxycholesterol and lag time (R=-0.31, P=0.0023). It is suggested that the higher 7 beta-hydroxycholesterol concentration in Lithuanian men is an indication of an increased in vivo lipid peroxidation.

Antioxidant state and mortality from coronary heart disease in Lithuanian and Swedish men: concomitant cross sectional study of men aged 50.

OBJECTIVE: To investigate possible risk factors and mechanisms behind the four times higher and diverging mortality from coronary heart disease in Lithuanian compared with Swedish middle aged men. DESIGN: Concomitant cross sectional comparison of randomly selected 50 year old men without serious acute or chronic disease. Methods and equipment were identical or highly standardised between the centres. SETTING: Linköping (Sweden) and Vilnius (Lithuania). SUBJECTS: 101 and 109 men aged 50 in Linköping and Vilnius respectively. MAIN OUTCOME MEASURES: Anthropometric data, blood pressure, smoking, plasma lipid and lipoprotein concentrations, susceptibility of low density lipoprotein to oxidation, and plasma concentrations of fat soluble antioxidant vitamins. RESULTS: Systolic blood pressure was higher (141 v 133 mm Hg, P < 0.01), smoking habits were similar, and plasma total cholesterol (5.10 v 5.49 mmol/l, P < 0.01) and low density lipoprotein cholesterol (3.30 v 3.68 mmol/l, P < 0.01) lower in men from Vilnius compared with those from Linköping. Triglyceride, high density lipoprotein cholesterol, and Lp(a) lipoprotein concentrations did not differ between the two groups. The resistance of low density lipoprotein to oxidation was lower in the men from Vilnius; lag phase was 67.6 v 79.5 minutes (P < 0.001). Also lower in the men from Vilnius were mean plasma concentrations of lipid soluble antioxidant vitamins (beta carotene 377 v 510 nmol/l, P < 0.01; lycopene 327 v 615 nmol/l, P < 0.001; and lipid adjusted gamma tocopherol 0.25 v 0.46 mumol/mmol, P < 0.001. alpha Tocopherol concentration did not differ). Regression analysis showed that the lag phase was still significantly shorter by 10 minutes in men from Vilnius when the influence of other known factors was taken into account. CONCLUSIONS: The high mortality from coronary heart disease in Lithuania is not caused by traditional risk factors alone. Mechanisms related to antioxidant state may be important.

Copper-induced low density lipoprotein oxidation is not a risk discriminator for intermittent claudication.

Oxidative modification of low density lipoprotein (LDL) is supposed to be important in atherogenesis. Recently it was shown that subjects with coronary atherosclerosis have an increased susceptibility of their LDL to copper-induced oxidation. We investigated if patients with intermittent claudication (IC) might have an increased susceptibility of LDL to copper-induced oxidation. Fifty-eight males were randomly selected from an epidemiological study of IC, 29 with IC and 29 healthy controls matched for age, sex and smoking habits. All subjects performed a standard exercise test to confirm or exclude peripheral atherosclerosis. Claudicants had a lag phase of 99.7 +/- 14.8 minutes (mean +/- SD) and in healthy controls it was 104.6 +/- 12.9 minutes. The difference between the groups was not significant and neither was there any association between lag phase and degree of peripheral atherosclerosis in IC. Lag phase showed a positive and significant correlation to the plasma concentration of high density lipoprotein-2 (HDL2-) cholesterol. The correlation for the whole group was r = 0.41, p < 0.01. We conclude that the susceptibility of LDL to copper-oxidation does not discriminate between claudicants and healthy controls. The results also suggest that high plasma concentrations of HDL2-cholesterol may have a protective effect on LDL against oxidation.

Effects of in vitro addition of captopril on copper-induced low density lipoprotein oxidation.

Low density lipoprotein (LDL) was incubated with 20 microM of the angiotensin converting enzyme (ACE) inhibitors captopril, fosinopril and quinapril or ethanol. Oxidation of LDL was initiated by addition of CuSO4 and monitored for production of conjugated dienes, thiobarbituric acid reactive substances (TBARS) and lipid peroxides. The inhibition of production of conjugated dienes was expressed as the lag phase in minutes. The lag phase for control samples was 55.2 +/- 6.1 (mean +/- s.e. mean) min and for captopril 86.4 +/- 7.0 min (P = 0.0058). Quinapril had a small but nonsignificant effect, fosinopril and ethanol had no effect. LDL incubated with captopril showed only 13.8% of TBARS and 22.7% of lipid peroxides produced by control (100%) after 1 h. Increasing concentrations of captopril showed a linear increase in the lag phase. We conclude that captopril increases the resistance of LDL to copper-induced oxidation.