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Journal of Clinical Endocrinology & Metabolism , doi:10.1210/jc.2004-0557
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The Journal of Clinical Endocrinology & Metabolism Vol. 90, No. 6 3786-3792
Copyright © 2005 by The Endocrine Society

REVIEW

Hepatic Lipase C-480T Genotype-Dependent Benefit from Long-Term Hormone Replacement Therapy for Atherosclerosis Progression in Postmenopausal Women

Yue-Mei Fan, Prasun Dastidar, Hannu Jokela, Reijo Punnonen and Terho Lehtimäki

Laboratory of Atherosclerosis Genetics (Y.-M.F., H.J., T.L.), Department of Clinical Chemistry, Center for Laboratory Medicine, University Hospital of Tampere and Tampere University Medical School; and Departments of Diagnostic Radiology (P.D.) and Obstetrics and Gynecology (R.P.), University Hospital of Tampere (P.D., R.P.) and Tampere University Medical School (P.D.), FIN-33521 Tampere, Finland

Address all correspondence and requests for reprints to: Yue-Mei Fan, Laboratory of Atherosclerosis Genetics, Department of Clinical Chemistry, Center for Laboratory Medicine, Tampere University Hospital, FinnMedi 2, 3rd Floor, P.O. Box 2000, FIN-33521 Tampere, Finland. E-mail: loyufa{at}uta.fi.


   Abstract
Top
 Abstract
Introduction
 Subjects and Methods
 Results
 Discussion
 References
 
Hepatic lipase (HL) is a lipolytic enzyme that hydrolyzes triglycerides and phospholipids in almost all major classes of lipoproteins. The HL gene has a functional promoter polymorphism at position –480, which affects transcription and leads to CC, CT, and TT genotypes. We investigated the effect of long-term hormone replacement therapy (HRT) on the progression of atherosclerosis in a 5-yr follow-up observational study of 88 postmenopausal women with different HL genotypes (CC, n = 49; CT, n = 34; TT, n = 5). These women, aged 45–71 yr, were divided into three groups based on the use of HRT. The HRT-EVP group (n = 26) used sequential estradiol valerate (EV) plus progestin (levonorgestrel), the HRT-EV group used EV alone (n = 32), and the control group (n = 30) used no HRT. The HRT-EV and HRT-EVP groups started estrogen at menopause for estrogen-deficiency symptoms, whereas the control group took no estrogen due to either the absence of such symptoms or a dislike of estrogen therapy. In addition to serum lipid concentration and HL genotype, the atherosclerosis severity score (ASC) for the abdominal aorta and carotid arteries was determined by ultrasonography. There was a significant interaction between HRT therapy and HL genotypes on the increase in ASC (P = 0.046) after adjustment for age, body mass index, changes in high-density lipoprotein cholesterol and baseline ASC. In subjects with the T allele, the progression of ASC was significantly faster in the control group than the HRT group (P = 0.0006), whereas in the CC genotype, there were no significant differences in ASC progression between the control and HRT groups. Our results suggest that the beneficial effect of HRT on atherosclerosis progression was restricted to women with the T allele, in whom the progression of ASC was slower by half. These results may help us understand in greater detail the benefits and possible risks associated with HRT in atherosclerotic diseases.


   Introduction
Top
 Abstract
 Introduction
Subjects and Methods
 Results
 Discussion
 References
 
CORONARY ATHEROSCLEROSIS IS an underlying cause of morbidity and mortality among women in the industrialized world. A number of observational studies have suggested that hormone replacement therapy (HRT) reduces the risk of atherosclerosis and coronary events in postmenopausal women (1, 2, 3, 4). However, recently published results from randomized clinical trials of HRT indicate that the therapy does not slow the progression of coronary atherosclerosis in postmenopausal women (5, 6, 7). As to why not all patients benefit and some are even harmed by such therapy, our understanding is limited. However, it is still possible that a genetically determined subgroup of the population could benefit from this therapy. Previous studies have demonstrated that subgroups with apolipoprotein (apo) E4 (–) (8), myeloperoxidase promoter –463 GG (9), or estrogen receptor 1 PvuII P/P genotype (10) are propitious for HRT. As part of an ongoing study of the genetic risk factors underlying variation in response to HRT, we focused on identifying genes predisposing to these differences.

One genetic candidate is a gene for hepatic lipase (HL), which is a lipolytic enzyme synthesized mostly by the liver (11). It has also recently been reported that HL is synthesized by macrophages (12). HL hydrolyzes triglycerides and phospholipids in almost all major classes of lipoproteins. It has also been held to act as a ligand that mediates the binding and uptake of lipoproteins via proteoglycans and/or receptor pathways (13). The HL gene has a functional promoter polymorphism at position C-480T (or 514) affecting transcription and leading to three genotypes (CC, CT, and TT). Previous studies have indicated that the presence of the T allele at position –480 is significantly consistently associated with low HL activity (14, 15, 16, 17). On the other hand, the significant association of this allele with elevated high-density lipoprotein (HDL) cholesterol levels is at odds with this (14, 15, 18, 19). HL activity appears to be regulated by several factors, including intraabdominal fat (20), sex steroid hormones (21, 22), and HL C-480T polymorphism (17, 23). HRT has been found to reduce postheparin plasma HL activity (24), which activity is inversely associated with endogenous estrogen levels (25) and is higher in postmenopausal compared with premenopausal women (26). Given the wide spectrum of the effects HL exerts on lipoprotein metabolism and the significance of the promoter variant, it is reasonable to hypothesize that genetic variation at this locus may also be involved in the variability in response to HRT treatment.

Two previous studies have investigated the relationship between the HL C-480T polymorphism and lipid profiles in postmenopausal women (27, 28). However, no previous study has reported on the relationship between the HL –480T polymorphism and progression of atherosclerosis severity in postmenopausal women during long-term HRT.

To elucidate the genetic variants that modulate HRT effects, we now assessed the association between the HL C-480T polymorphism and the effect of HRT in response of atherosclerosis severity among postmenopausal women. The purpose was to establish whether, as hypothesized, the HL C-480T polymorphism modifies the effect of HRT on the development of atherosclerosis.


   Subjects and Methods
Top
 Abstract
 Introduction
 Subjects and Methods
Results
 Discussion
 References
 
Subjects

In 1993 women attending a private outpatient clinic in Tampere for annual routine gynecological examinations were invited to participate. For the cross-sectional baseline study in 1993 (3), 120 nonsmoking, nondiabetic postmenopausal women aged 45–71 yr were enrolled. Eighty-eight of these women participated in this 5-yr follow-up study from 1993 to 1998. They had no clinically evident cardiovascular diseases or hypertension and were classified into three groups based on the use of HRT. The HRT-EVP group (n = 26) used estradiol valerate (EV) at 2 mg/d for 11 d, followed by EV continued with progestin (P) (levonorgestrel, 0.25 mg/d) for 10 d. In the HRT-EV group (n = 32), the treatment was EV at 2 mg/d continuously; the control group (n = 30) had never used HRT. In the HRT-EVP and HRT-EV groups, there was a pause in therapy for 7 d after each 21-d cycle. None of these women discontinued the therapy during follow-up. HRT, when used, was started at the time of menopause for climacteric symptoms. In the control group the main reasons for nonuse of HRT was the absence of vasomotor and other climacteric symptoms and dislike of HRT. In the HRT-EV, HRT-EVP, and control groups, 24, four, and six women had undergone hysterectomy, respectively. At baseline, the mean duration of EV and EVP treatment was 9.2 ± 3.7 and 10.9 ± 2.5 yr, respectively. The mean time from menopause in the control group was 11.9 ± 4.0 yr. The mean ages in the HRT-EVP, HRT-EV, and control groups were 59.7 ± 5.5, 60.4 ± 4.8, and 61.5 ± 5.8 yr, respectively (P = 0.441, by ANOVA). The mean body mass index (BMI) was similar in all study groups (P = 0.953). At baseline, all women were clinically healthy and used no lipid-lowering or other chronic medication. Ultrasonography and blood sampling took place in the University Hospital of Tampere. The study was approved by the ethics committee of the hospital. All participants signed an informed consent document.

Blood samples

Blood samples for serum lipid and genotype analyses were taken after overnight fasting. Sampling took place within 3 wk from ultrasonography and for HRT-EVP users during the third week of the hormone regimen. After separation of serum by low-speed centrifugation, the sera were divided into aliquots and stored at –70 C until analyzed.

Ultrasonography

Ultrasonography at baseline and follow-up were performed with Sonolayer V SSA 100 equipment (Toshiba Corp., Tokyo, Japan), as reported in detail elsewhere (3, 10). In brief, transverse and longitudinal scans of the extracranial carotid arteries were made bilaterally at four different segments of the carotid. Only fibrous and calcified atherosclerotic lesions were taken into consideration and were defined as plaques when distinct areas of mineralization and/or focal protrusion into the lumen were identified. An intimal-media far-wall thickness equal to or more than 1.3 mm at any carotid artery segment was defined as an atherosclerotic plaque (29) and the total number of plaques (NAP) was calculated. All carotid artery examinations were made with a 5.0-MHz convex transducer probe.

Longitudinal ultrasonographs of the abdominal aorta were obtained at 1-cm intervals and transverse scans at 2-cm intervals in the area of three aortic segments. Significant aortic plaques were defined as an intima-media far-wall thickness equal to or more than 3.0 mm (29). All aortic examinations were performed with a 3.75-MHz convex transducer probe. The reproducibility of our ultrasonographic protocol for significant aortic and carotid plaques was also examined: 1 month after the first assessment, 20 randomly selected subjects were invited to attend for a repeat examination. The repeatability of NAP between the first and second examination was 90% for the carotid artery segment areas and 100% for the aortic segments. All ultrasonographies were performed in blinded manner by one experienced ultrasonographer and radiologist (P.D.).

The atherosclerosis severity score (ASC) was constructed by dividing the atherosclerosis in the abdominal aorta and carotid arteries into three severity classes, i.e. 1 = slight (1.3–2 mm), 2 = moderate (2–3 mm), and 3 = severe (more than 3 mm). The ASC was then calculated as the sum of the severity classes in aorta and carotid artery. The total NAP was calculated (in the baseline study only because 5-yr data were not available) according to the NAP (criteria for plaques as above). Scoring was done by one person (P.D.) in blinded manner, without knowledge of HRT and HL genotype status.

HL genotyping

DNA was isolated from white blood cells using a commercial kit (QIAGEN Inc., Valencia, CA). As described previously (30), determination of the HL promoter C-480T genotype was carried out by PCR amplification using the primers 5'-AAGAAGTGTGTTTACTCTAGGATCA-3' and 5'-GGTGGCTTCCACGTGGCTGCCTAAG-3'. Thermal cycling conditions were initial denaturation at 96 C for 3 min, followed by 33 cycles of amplification at 96 C for 1 min, annealing at 65.5 C for 1 min, and extension at 72 C for 1 min, with a final extension at 72 C for 5 min. The amplified DNA fragments were digested with the restriction enzyme NlaIII (New England Biolabs, Beverly, MA) followed by electrophoresis on 3% agarose gel.

Other laboratory analyses

Lipid measurements were made at baseline and after the 5-yr follow-up. Serum total cholesterol and triglycerides were determined by a commercial method (Kodak Echtachem 700 XR; Eastman Kodak Co., Clinical Products Division, Rochester, NY). Serum HDL cholesterol and its subfractions (HDL2 and HDL3) were separated by a dextran-sulfate-magnesium precipitation procedure, and the cholesterol content was analyzed with a Monarch 2000 analyzer (Instrumentation Laboratory, Lexington, KY), using the cholesterinoxidase-peroxidase/antiperoxidase cholesterol reagent (catalog no. 237574; Roche, Mannheim, Germany) and a primary cholesterol standard (catalog no. 530, Orion Diagnostics, Helsinki, Finland). Low-density lipoprotein (LDL) cholesterol content was calculated according to the Friedewald formula (31). Apolipoproteins A1 and B were determined on a Monarch analyzer by an immunoturbidimetric method (32) (catalog no. 67265 and 67249, Orion Diagnostics). In the follow-up study, all lipid analyses were performed with a Cobas Integra 700 automatic analyzer with the reagents and calibrators recommended by the manufacturer (Roche Diagnostics, Basel, Switzerland).

Statistical analyses

Differences in clinical characteristics and concentrations of lipids and lipoproteins between the HL C-480T subgroups within the HRT and control groups were measured by one-way analysis of covariance (ANCOVA), using age and BMI as covariates when appropriate. Because the triglycerides concentrations had a skewed distribution, the statistical analyses were based on log-transformed data. The triglycerides concentrations in the tables are nevertheless given as crude values. Changes in lipid concentrations, lipoprotein concentrations, and atherosclerosis severity during follow-up were expressed as mean (± SD), and differences within the treatment groups were tested by ANCOVA, with age and BMI as covariates.

Interaction between the HL genotype and treatment (control or HRT) was tested by two-way ANCOVA. We tested whether the interaction between genotype and treatment group was independent of changes in HDL cholesterol during the trial, baseline ASC, age, and BMI, using these as covariates in two-way ANCOVA.

Longitudinal follow-up data were analyzed by ANOVA for repeated measures (RANOVA) to find interactions between treatment groups (HRT vs. control) and time points. RANOVA was performed separately within the HL genotype groups using the least significant differences post hoc test to determine the significance of differences between treatment groups in ASC and NAP at baseline. A similar analysis was made in ASC at follow-up. All analyses were performed with the Statistica for Windows version 5.1 software package (Statsoft, Inc., Tulsa, OK). The level of significance was set at P < 0.05.


   Results
Top
 Abstract
 Introduction
 Subjects and Methods
 Results
Discussion
 References
 
In the total cohort, the C and T alleles were found at frequencies of 0.75 and 0.25, respectively; 49 (55.7%) had the CC genotype, 34 (38.6%) the CT, and 5 (5.7%) the TT genotype. In the HRT-EV group, the CC genotype was found in 16 subjects (50%), CT in 14 subjects (43.8%), and TT genotype in two subjects (6.3%). In the HRT-EVP group, the CC genotype was observed in 16 subjects (61.5%) and the CT genotype was observed in 10 subjects (38.5%). In the control group, the corresponding data for CC, CT, and TT genotype were 17 (56.7%), 10 (33.3%), and three (10%). These frequencies did not differ significantly among the three groups. Because there were no TT homozygous subjects in the HRT-EVP group and there was no statistically significant difference between the CT and TT genotype groups in any baseline characteristics, the T allele carriers were combined into one group, and this was compared with subjects with the CC homozygous genotype.

The baseline distribution of known characteristics among postmenopausal women with respect to treatment and HL C-480T genotype status is presented in Table 1Go. Among the total, CC genotype carriers were older than women with the T allele (P = 0.01). There was an association of this allele with high levels of HDL3 cholesterol in all subjects (P = 0.04). When the subjects were classified according to their C-480T genotype, there were no statistically significant differences between groups at baseline in total cholesterol, triglycerides, LDL cholesterol, apo AI, apo B, HDL, HDL2, and HDL3 cholesterol (Table 1Go). In the HRT-EV group, the CC genotype carriers were older than women with the T allele (P < 0.001).


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  		TABLE 1. Serum lipids and apolipoproteins in postmenopausal women at baseline by HRT and HL genotype status

 
At baseline, ASCs in the HRT-EV, HRT-EVP, and controls were 1.13, 1.31, and 1.59, respectively, in subjects with the CC genotype. There was no significant treatment effect on ASC (ANCOVA for trend, P = 0.282, adjusted by age and BMI; Fig. 1AGo). After the 5-yr follow-up, the corresponding differences between the HRT-EV and HRT-EVP groups and controls were 2.94, 3.31, and 3.71, respectively (ANCOVA for trend, P = 0.266, adjusted by age and BMI; Fig. 1BGo). At baseline, CC genotype carriers in HRT-EV had a mean of 45.9% (1.75 vs. 3.24 in controls, P = 0.006) lower total NAP. There was, however, no significant treatment effect on NAP between HRT-EVP and controls (2.44 vs. 3.24, P = 0.131) in CC carriers (ANCOVA for trend, P = 0.021, adjusted by age and BMI).



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  		FIG. 1. A–D, ASC in postmenopausal women by HRT group and HL genotype status (CC and CT, TT). A and B, Results from the baseline study. C and D, Results from the cross-sectional study after 5-yr follow-up. The P values for the mean (± SD, whiskers) differences between the HRT groups and controls shown in the figure were obtained by ANCOVA, with least significance test as post hoc test. Results are adjusted for age and BMI.

 
At baseline, subjects with the T allele in HRT-EV tended to have an average 45.8% lower ASC (1.13 vs. 2.08 in controls, P = 0.005), and those in HRT-EVP had a 37.4% lower ASC (1.30 vs. 2.08 in controls, P = 0.037) than the controls (ANCOVA for trend, P = 0.014, adjusted by age and BMI; Fig. 1CGo). After the 5-yr follow-up, the corresponding differences between the HRT-EV and HRT-EVP groups and controls were 43.3% (2.88 vs. 5.08, P = 0.0003) and 40.9% (3.00 vs. 5.08, P = 0.002; ANCOVA for trend, P = 0.001, adjusted for age and BMI; Fig. 1DGo). At baseline, subjects with the T allele in HRT-EV had a mean 60.7% (1.88 vs. 4.77 in controls, P = 0.0003) lower total NAP, and subjects in HRT-EVP a 43.4% (2.70 vs. 4.77 in controls, P = 0.015) lower total NAP than the controls (ANCOVA for trend, P = 0.002, adjusted by age and BMI).

In subsequent analyses, we combined the data on use of EV and EVP because the results for the two therapies were similar. The HL C-480T polymorphism had no effect on changes in total cholesterol, LDL cholesterol, triglycerides, and HDL cholesterol concentrations in the two study groups. HRT reduced total cholesterol and LDL cholesterol and increased HDL cholesterol and triglycerides to a similar extent in the two genotype groups (Table 2Go). There were no statistically significant differences between HL genotypes in response of the studied lipid traits (HL genotype group by time point interaction not significant for all lipids within study groups).


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  		TABLE 2. Influence of HL genotypes on the baseline and follow-up lipid levels and ASC in the HRT and control groups

 
A significant interaction between the HL C-480T genotype and HRT in respect of increase in ASC was observed (P = 0.046) after adjustment for age, BMI, changes in HDL cholesterol during the trial, and ASC at baseline. A larger increase in ASC indicates greater progression of atherosclerosis. The benefit of HRT was restricted to women with the T allele. Among subjects with the CC genotype, the progression rate of ASC in HRT users and controls did not differ statistically significantly (treatment group by time point interaction in RANOVA, P = 0.618, Fig. 2AGo). Among the T allele carriers, the ASC progression rate differed significantly between HRT users and controls (treatment group by time point interaction in RANOVA, P = 0.0006, Fig. 2BGo).



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  		FIG. 2. A and B, The effect of HRT on the progression of atherosclerosis, as measured by ASC in postmenopausal women with CC genotype (A) and CT or TT genotype (B), compared with the progression in controls with the same HL genotype and time elapsed from menopause but without HRT. The P values shown in the figure are from two-way ANOVA (grouping HRT/control) for repeated measures.

 
The corresponding changes in serum lipid and lipoproteins and changes in ASC during follow-up, according to HL genotype and treatment status, are shown in Table 2Go. There were no statistically significant differences between HL genotypes in changes in lipid and lipoprotein concentrations. Among control women, the T allele was associated with greater changes in ASC during follow-up than the CC genotype (P = 0.04) (Table 2Go). The association of the T allele with more substantial progression of atherosclerosis, as observed in the control group, was influenced by the use of HRT. In fact, the T allele appeared to be associated with lesser progression in women who were receiving HRT. In these women, the progression of atherosclerosis severity was slower by half in those receiving HRT, whereas no difference was observed in women carrying the CC genotype.


   Discussion
Top
 Abstract
 Introduction
 Subjects and Methods
 Results
 Discussion
References
 
The present study assessed the role of the HL C-480T polymorphism and long-term HRT in atherosclerosis severity in postmenopausal women. The T allele was found to be associated with greater changes in ASC during follow-up than the CC genotype among control women. However, T allele carriers were seen to reflect beneficial effects on atherosclerosis progression during long-term HRT. The effect of the T allele on ASC with HRT during follow-up was independent of HDL cholesterol. No positive effect of HRT was seen in the CC genotype group, in which the progression of atherosclerosis tended to be similar among HRT users and controls during the 5-yr follow-up. Moreover, we found a significant interaction between the HL C-480T polymorphism and HRT in respect of atherosclerosis progression. We have thus revealed a genetic predisposition for response to HRT.

In this study, all subjects were nonsmokers; did not have diabetes; were without clinically evident cardiovascular disease, hypertension, or chronic medication; and were otherwise clinically healthy. In addition, dietary analysis revealed no substantial differences in the use of saturated fat or dietary cholesterol between the HRT groups (3); our results thus are unlikely to have been influenced by differences in dietary habits between the HRT and control groups. Because some other factors possibly differing between users and nonusers (e.g. socioeconomic status) were not accounted for, it is conceivable that some unknown factors may have biased our results.

Several observational studies have suggested that HRT reduces the risk of atherosclerosis and coronary events in postmenopausal women (1, 2, 3, 4). However, an adverse effect appears in the first year of HRT use, decreasing over time (6). It has been reported that HRT has beneficial effects in the early stages of atherosclerosis progression but is of little or no benefit in advanced stages of the disease (33). In our study, the plaques detected by ultrasonography were mostly uncalcified, constituting less advanced atherosclerotic lesions. Recent studies from our group (8, 9, 10) have shown that the effect of HRT varies among individuals and that this variation is partly genetically determined by functional variations in candidate genes. Together with our finding, it is becoming increasingly clear that the individual response to treatment is related to a great extent to genetically defined differences.

Zambon et al. (34) found HL genotype differences in effect on coronary stenosis progression in the HL genotype order of CC < CT < TT during intensive lipid medication. However, the design in the study in question differed from that here. In our study we also included a control group without medication in addition to the HRT treatment group, which allowed us first to test HL genotype-by-treatment group (control/HRT) interaction, which was statistically significant, and thereafter compare the effects in treated vs. nontreated groups on ASC progression in different genotype groups. Zambon et al. did not include a control group in their statistical analysis, and it is thus difficult to know which part of the changes during intervention including extensive lipid-lowering therapy was due to medication or alternatively other lifestyle changes during the same period. Thus, the differences in study design and medications used make comparison of the results from these two studies very difficult.

In our study, the HL promoter polymorphism had no effect on the lipid response to HRT in postmenopausal women, as previously reported (27, 28). Because HL is expressed within the macrophages of the atherosclerosis plaque (12, 35), it has been postulated that HL might have a direct role in the pathogenesis of atherosclerosis through a pathway not involving changes in plasma lipoprotein metabolism (12, 35). The C-480T polymorphism is a key determinant of HL activity, accounting for up to 38% of its variability (36). HL activity is also hormonally affected. Estrogen replacement therapy in postmenopausal women acutely reduces HL activity (24, 37, 38). Tikkanen et al. (25) have shown that HL activity falls significantly in healthy premenopausal women during the luteal phase of the menstrual cycle, when endogenous estradiol levels are highest. HL activity decreases progressively throughout pregnancy as estradiol levels increase (39). The fall in endogenous estrogen with menopause, again, is associated with a rise in HL activity (26, 40). The effect of the HL C-480T polymorphism on coronary heart disease (CHD) has been sought in several studies. The T allele is associated with endothelial dysfunction in young healthy men (30). This allele is also associated with coronary artery calcification (41), the extent of coronary stenosis in patients with CHD (42), and prevalent CHD case-control status in men (14, 43). On the other hand, Rundek et al. (44) reported that the CC genotype is associated with greater intimal-media thickness than in T allele carriers. A group under Faggin (45) showed the CC genotype to be associated with an abundance of macrophages in patients with severe carotid artery stenosis undergoing carotid endarterectomy. Thus, the relationship between the HL C-480T polymorphism and CHD is not consistent. Moreover, there is evidence for an interaction of this polymorphism with dietary fat intake (46), lipid-lowering medications (34), physical activity (47), and HDL cholesterol level (48). Our finding may be related to the HL activity either in the liver or alternatively in artery wall macrophages (12, 35). The exact mechanism remains to be established.

Ultrasonographic methods permit evaluation of atherosclerosis in asymptomatic subjects (49, 50). In our study ASC represents the severity of the plaques found in the large arteries, i.e. carotids and aorta. NAP represents only the number of total plaques found in the above-mentioned regions. The number of plaques may be substantial but if they are not large enough to cause stenosis of the artery, they will not induce severe clinical symptoms. Because ASC represents the size and severity of plaques, we feel that ASC is a better marker of the severity of atherosclerosis. Because individuals with more advanced atherosclerotic disease, characterized by stenotic or occlusive lesions, were not included here, the association of the HL genotype with atherosclerosis severity status may have been modified. Likewise, because we did not study the cardiovascular end process, we cannot rule out selection bias related to HRT or certain other noncausal explanations for our findings.

Our results suggest that the C-480T polymorphism confers a genotype-specific responsiveness to HRT in atherosclerosis progression in postmenopausal women. The findings may help us understand in greater detail the benefit and possible risk of HRT in atherosclerotic diseases. In the future, identification and characterization of the genotype might help patients and physicians assess the risks and benefits of HRT and lead to new insights into HRT action.


   Acknowledgments
 
We thank Miss Nina Peltonen for her skillful technical assistance.


   Footnotes
 
This work was supported by grants from the Finnish Foundation of Cardiovascular Research, the Emil Aaltonen Foundation, the Medical Research Fund of Tampere University Hospital, the Research Foundation of Orion Corp., the Pirkanmaa Regional Fund of the Finnish Cultural Foundation, the Ida Montin Foundation, and the Academy of Finland (Grant 104821).

First Published Online March 8, 2005

Abbreviations: ANCOVA, Analysis of covariance; apo, apolipoprotein; ASC, atherosclerosis severity score; BMI, body mass index; CHD, coronary heart disease; EV, estradiol valerate; EVP, EV and progestin; HDL, high-density lipoprotein; HL, hepatic lipase; HRT, hormone replacement therapy; LDL, low-density lipoprotein; NAP, total number of plaques; RANOVA, ANOVA for repeated measures.

Received March 23, 2004.

Accepted March 2, 2005.


   References
Top
 Abstract
 Introduction
 Subjects and Methods
 Results
 Discussion
 References
 

  1. Bush TL, Barrett-Connor E, Cowan LD, Criqui MH, Wallace RB, Suchindran CM, Tyroler HA, Rifkind BM 1987 Cardiovascular mortality and noncontraceptive use of estrogen in women: results from the Lipid Research Clinics Program Follow-Up Study. Circulation 75:1102–1109[Abstract/Free Full Text]
  2. Nabulsi AA, Folsom AR, White A, Patsch W, Heiss G, Wu KK, Szklo M 1993 Association of hormone-replacement therapy with various cardiovascular risk factors in postmenopausal women. The Atherosclerosis Risk in Communities Study Investigators. N Engl J Med 328:1069–1075[Abstract/Free Full Text]
  3. Punnonen RH, Jokela HA, Dastidar PS, Nevala M, Laippala PJ 1995 Combined oestrogen-progestin replacement therapy prevents atherosclerosis in postmenopausal women. Maturitas 21:179–187[CrossRef][Medline]
  4. Stampfer MJ, Colditz GA, Willett WC, Manson JE, Rosner B, Speizer FE, Hennekens CH 1991 Postmenopausal estrogen therapy and cardiovascular disease. Ten-year follow-up from the Nurses’ Health Study. N Engl J Med 325:756–762[Abstract]
  5. Herrington DM, Reboussin DM, Brosnihan KB, Sharp PC, Shumaker SA, Snyder TE, Furberg CD, Kowalchuk GJ, Stuckey TD, Rogers WJ, Givens DH, Waters D 2000 Effects of estrogen replacement on the progression of coronary-artery atherosclerosis. N Engl J Med 343:522–529[Abstract/Free Full Text]
  6. Grady D, Herrington D, Bittner V, Blumenthal R, Davidson M, Hlatky M, Hsia J, Hulley S, Herd A, Khan S, Newby LK, Waters D, Vittinghoff E, Wenger N 2002 Cardiovascular disease outcomes during 6.8 years of hormone therapy: Heart and Estrogen/Progestin Replacement Study follow-up (HERS II). JAMA 288:49–57[Abstract/Free Full Text]
  7. Rossouw JE, Anderson GL, Prentice RL, LaCroix AZ, Kooperberg C, Stefanick ML, Jackson RD, Beresford SA, Howard BV, Johnson KC, Kotchen JM, Ockene J 2002 Risks and benefits of estrogen plus progestin in healthy postmenopausal women: principal results from the Women’s Health Initiative randomized controlled trial. JAMA 288:321–333[Abstract/Free Full Text]
  8. Lehtimaki T, Dastidar P, Jokela H, Koivula T, Lehtinen S, Ehnholm C, Punnonen R 2002 Effect of long-term hormone replacement therapy on atherosclerosis progression in postmenopausal women relates to functional apolipoprotein e genotype. J Clin Endocrinol Metab 87:4147–4153[Abstract/Free Full Text]
  9. Makela R, Dastidar P, Jokela H, Saarela M, Punnonen R, Lehtimaki T 2003 Effect of long-term hormone replacement therapy on atherosclerosis progression in postmenopausal women relates to myeloperoxidase promoter polymorphism. J Clin Endocrinol Metab 88:3823–3828[Abstract/Free Full Text]
  10. Koivu TA, Fan YM, Mattila KM, Dastidar P, Jokela H, Nikkari ST, Kunnas T, Punnonen R, Lehtimaki T 2003 The effect of hormone replacement therapy on atherosclerotic severity in relation to ESR1 genotype in postmenopausal women. Maturitas 44:29–38[CrossRef][Medline]
  11. Sanan DA, Fan J, Bensadoun A, Taylor JM 1997 Hepatic lipase is abundant on both hepatocyte and endothelial cell surfaces in the liver. J Lipid Res 38:1002–1013[Abstract]
  12. Gonzalez-Navarro H, Nong Z, Freeman L, Bensadoun A, Peterson K, Santamarina-Fojo S 2002 Identification of mouse and human macrophages as a site of synthesis of hepatic lipase. J Lipid Res 43:671–675[Abstract/Free Full Text]
  13. Santamarina-Fojo S, Haudenschild C, Amar M 1998 The role of hepatic lipase in lipoprotein metabolism and atherosclerosis. Curr Opin Lipidol 9:211–219[CrossRef][Medline]
  14. Jansen H, Verhoeven AJ, Weeks L, Kastelein JJ, Halley DJ, van den Ouweland A, Jukema JW, Seidell JC, Birkenhager JC 1997 Common C-to-T substitution at position –480 of the hepatic lipase promoter associated with a lowered lipase activity in coronary artery disease patients. Arterioscler Thromb Vasc Biol 17:2837–2842[Abstract/Free Full Text]
  15. Tahvanainen E, Syvanne M, Frick MH, Murtomaki-Repo S, Antikainen M, Kesaniemi YA, Kauma H, Pasternak A, Taskinen MR, Ehnholm C 1998 Association of variation in hepatic lipase activity with promoter variation in the hepatic lipase gene. The LOCAT Study Investigators. J Clin Invest 101:956–960[Medline]
  16. Carr MC, Hokanson JE, Deeb SS, Purnell JQ, Mitchell ES, Brunzell JD 1999 A hepatic lipase gene promoter polymorphism attenuates the increase in hepatic lipase activity with increasing intra-abdominal fat in women. Arterioscler Thromb Vasc Biol 19:2701–2707[Abstract/Free Full Text]
  17. Vega GL, Clark LT, Tang A, Marcovina S, Grundy SM, Cohen JC 1998 Hepatic lipase activity is lower in African American men than in white American men: effects of 5' flanking polymorphism in the hepatic lipase gene (LIPC). J Lipid Res 39:228–232[Abstract/Free Full Text]
  18. Murtomaki S, Tahvanainen E, Antikainen M, Tiret L, Nicaud V, Jansen H, Ehnholm C 1997 Hepatic lipase gene polymorphisms influence plasma HDL levels. Results from Finnish EARS participants. European Atherosclerosis Research Study. Arterioscler Thromb Vasc Biol 17:1879–1884[Abstract/Free Full Text]
  19. Hegele RA, Harris SB, Brunt JH, Young TK, Hanley AJ, Zinman B, Connelly PW 1999 Absence of association between genetic variation in the LIPC gene promoter and plasma lipoproteins in three Canadian populations. Atherosclerosis 146:153–160[CrossRef][Medline]
  20. Despres JP, Ferland M, Moorjani S, Nadeau A, Tremblay A, Lupien PJ, Theriault G, Bouchard C 1989 Role of hepatic-triglyceride lipase activity in the association between intra-abdominal fat and plasma HDL cholesterol in obese women. Arteriosclerosis 9:485–492[Abstract/Free Full Text]
  21. Ehnholm C, Huttunen JK, Kinnunen PJ, Miettinen TA, Nikkila EA 1975 Effect of oxandrolone treatment on the activity of lipoprotein lipase, hepatic lipase and phospholipase A1 of human postheparin plasma. N Engl J Med 292:1314–1317[Abstract]
  22. Colvin Jr PL, Auerbach BJ, Case LD, Hazzard WR, Applebaum-Bowden D 1991 A dose-response relationship between sex hormone-induced change in hepatic triglyceride lipase and high-density lipoprotein cholesterol in postmenopausal women. Metabolism 40:1052–1056[CrossRef][Medline]
  23. Zambon A, Deeb SS, Hokanson JE, Brown BG, Brunzell JD 1998 Common variants in the promoter of the hepatic lipase gene are associated with lower levels of hepatic lipase activity, buoyant LDL, and higher HDL2 cholesterol. Arterioscler Thromb Vasc Biol 18:1723–1729[Abstract/Free Full Text]
  24. Brinton EA 1996 Oral estrogen replacement therapy in postmenopausal women selectively raises levels and production rates of lipoprotein A-I and lowers hepatic lipase activity without lowering the fractional catabolic rate. Arterioscler Thromb Vasc Biol 16:431–440[Abstract/Free Full Text]
  25. Tikkanen MJ, Kuusi T, Nikkila EA, Sipinen S 1986 Post-menopausal hormone replacement therapy: effects of progestogens on serum lipids and lipoproteins. A review. Maturitas 8:7–17[CrossRef][Medline]
  26. Ikenoue N, Wakatsuki A, Okatani Y 1999 Small low-density lipoprotein particles in women with natural or surgically induced menopause. Obstet Gynecol 93:566–570[CrossRef][Medline]
  27. Somekawa Y, Umeki H, Kobayashi K, Tomura S, Aso T, Hamaguchi H 2002 Effects of hormone replacement therapy and hepatic lipase polymorphism on serum lipid profiles in postmenopausal Japanese women. J Clin Endocrinol Metab 87:4766–4770[Abstract/Free Full Text]
  28. Yamakawa-Kobayashi K, Somekawa Y, Fujimura M, Tomura S, Arinami T, Hamaguchi H 2002 Relation of the –514C/T polymorphism in the hepatic lipase gene to serum HDL and LDL cholesterol levels in postmenopausal women under hormone replacement therapy. Atherosclerosis 162:17–21[CrossRef][Medline]
  29. Furberg CD, Byington RP, Borhani NA 1989 Multicenter isradipine diuretic atherosclerosis study (MIDAS). Design features. The Midas Research Group. Am J Med 86:37–39[CrossRef][Medline]
  30. Fan Y, Laaksonen R, Janatuinen T, Vesalainen R, Nuutila P, Koivula T, Knuuti J, Lehtimaki T 2001 Hepatic lipase gene variation is related to coronary reactivity in healthy young men. Eur J Clin Invest 31:574–580[CrossRef][Medline]
  31. Friedewald WT, Levy RI, Fredrickson DS 1972 Estimation of the concentration of low-density lipoprotein cholesterol in plasma, without use of the preparative ultracentrifuge. Clin Chem 18:499–502[Abstract]
  32. Riepponen P, Marniemi J, Rautaoja T 1987 Immunoturbidimetric determination of apolipoproteins A-1 and B in serum. Scand J Clin Lab Invest 47:739–744[Medline]
  33. Mikkola TS, Clarkson TB 2002 Estrogen replacement therapy, atherosclerosis, and vascular function. Cardiovasc Res 53:605–619[Abstract/Free Full Text]
  34. Zambon A, Deeb SS, Brown BG, Hokanson JE, Brunzell JD 2001 Common hepatic lipase gene promoter variant determines clinical response to intensive lipid-lowering treatment. Circulation 103:792–798[Abstract/Free Full Text]
  35. Nong Z, Gonzalez-Navarro H, Amar M, Freeman L, Knapper C, Neufeld EB, Paigen BJ, Hoyt RF, Fruchart-Najib J, Santamarina-Fojo S 2003 Hepatic lipase expression in macrophages contributes to atherosclerosis in apoE-deficient and LCAT-transgenic mice. J Clin Invest 112:367–378[CrossRef][Medline]
  36. De Oliveira e Silva ER, Kong M, Han Z, Starr C, Kass EM, Juo SH, Foster D, Dansky HM, Merkel M, Cundey K, Brinton EA, Breslow JL, Smith JD 1999 Metabolic and genetic determinants of HDL metabolism and hepatic lipase activity in normolipidemic females. J Lipid Res 40:1211–1221[Abstract/Free Full Text]
  37. Tikkanen MJ, Nikkila EA, Kuusi T, Sipinen S 1982 Effects of oestradiol and levonorgestrel on lipoprotein lipids and postheparin plasma lipase activities in normolipoproteinaemic women. Acta Endocrinol (Copenh) 99:630–635[Abstract/Free Full Text]
  38. Applebaum DM, Goldberg AP, Pykalisto OJ, Brunzell JD, Hazzard WR 1977 Effect of estrogen on post-heparin lipolytic activity. Selective decline in hepatic triglyceride lipase. J Clin Invest 59:601–608
  39. Alvarez JJ, Montelongo A, Iglesias A, Lasuncion MA, Herrera E 1996 Longitudinal study on lipoprotein profile, high density lipoprotein subclass, and postheparin lipases during gestation in women. J Lipid Res 37:299–308[Abstract]
  40. Berg GA, Siseles N, Gonzalez AI, Ortiz OC, Tempone A, Wikinski RW 2001 Higher values of hepatic lipase activity in postmenopause: relationship with atherogenic intermediate density and low density lipoproteins. Menopause 8:51–57[CrossRef][Medline]
  41. Hokanson JE, Cheng S, Snell-Bergeon JK, Fijal BA, Grow MA, Hung C, Erlich HA, Ehrlich J, Eckel RH, Rewers M 2002 A common promoter polymorphism in the hepatic lipase gene (LIPC-480C>T) is associated with an increase in coronary calcification in type 1 diabetes. Diabetes 51:1208–1213[Abstract/Free Full Text]
  42. Dugi KA, Brandauer K, Schmidt N, Nau B, Schneider JG, Mentz S, Keiper T, Schaefer JR, Meissner C, Kather H, Bahner ML, Fiehn W, Kreuzer J 2001 Low hepatic lipase activity is a novel risk factor for coronary artery disease. Circulation 104:3057–3062[Abstract/Free Full Text]
  43. Ji J, Herbison CE, Mamotte CD, Burke V, Taylor RR, van Bockxmeer FM 2002 Hepatic lipase gene –514 C/T polymorphism and premature coronary heart disease. J Cardiovasc Risk 9:105–113[CrossRef][Medline]
  44. Rundek T, Elkind MS, Pittman J, Boden-Albala B, Martin S, Humphries SE, Juo SH, Sacco RL 2002 Carotid intima-media thickness is associated with allelic variants of stromelysin-1, interleukin-6, and hepatic lipase genes: the Northern Manhattan Prospective Cohort Study. Stroke 33:1420–1423[Abstract/Free Full Text]
  45. Faggin E, Zambon A, Puato M, Deeb SS, Bertocco S, Sartore S, Crepaldi G, Pessina AC, Pauletto P 2002 Association between the –514 C–>T polymorphism of the hepatic lipase gene promoter and unstable carotid plaque in patients with severe carotid artery stenosis. J Am Coll Cardiol 40:1059–1066[Abstract/Free Full Text]
  46. Ordovas JM, Corella D, Demissie S, Cupples LA, Couture P, Coltell O, Wilson PW, Schaefer EJ, Tucker KL 2002 Dietary fat intake determines the effect of a common polymorphism in the hepatic lipase gene promoter on high-density lipoprotein metabolism: evidence of a strong dose effect in this gene-nutrient interaction in the Framingham Study. Circulation 106:2315–2321[Abstract/Free Full Text]
  47. Hokanson JE, Kamboh MI, Scarboro S, Eckel RH, Hamman RF 2003 Effects of the hepatic lipase gene and physical activity on coronary heart disease risk. Am J Epidemiol 158:836–843[Abstract/Free Full Text]
  48. Fan YM, Salonen JT, Koivu TA, Tuomainen TP, Nyyssonen K, Lakka TA, Salonen R, Seppanen K, Nikkari ST, Tahvanainen E, Lehtimäki T 2004 Hepatic lipase C-480T polymorphism modifies the effect of HDL cholesterol on the risk of acute myocardial infarction in men: a prospective population based study. J Med Genet 41:E28
  49. Cheng KS, Mikhailidis DP, Hamilton G, Seifalian AM 2002 A review of the carotid and femoral intima-media thickness as an indicator of the presence of peripheral vascular disease and cardiovascular risk factors. Cardiovasc Res 54:528–538[Abstract/Free Full Text]
  50. Howard G, Sharrett AR, Heiss G, Evans GW, Chambless LE, Riley WA, Burke GL 1993 Carotid artery intimal-medial thickness distribution in general populations as evaluated by B-mode ultrasound. ARIC Investigators. Stroke 24:1297–1304[Abstract/Free Full Text]




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