Abnormal elevation of low-density lipoprotein (LDL) and triglyceride-rich lipoproteins in plasma as well as dysfunction of anti-atherogenic high-density lipoprotein (HDL) have both been recognized as essential components of the pathogenesis of atherosclerosis and are classified as dyslipidemia. This review describes the arc of development of antisense oligonucleotides for the treatment of dyslipidemia. Chemically-armed antisense candidates can act on various kinds of transcripts, including mRNA and miRNA, via several different endogenous antisense mechanisms, and have exhibited potent systemic anti-dyslipidemic effects. Here, we present specific cutting-edge technologies have recently been brought into antisense strategies, and describe how they have improved the potency of antisense drugs in regard to pharmacokinetics and pharmacodynamics. In addition, we discuss perspectives for the use of armed antisense oligonucleotides as new clinical options for dyslipidemia, in the light of outcomes of recent clinical trials and safety concerns indicated by several clinical and preclinical studies.

Aim: Although arterial stiffness has been associated with the development of atherosclerosis, the role of brachial-ankle pulse wave velocity (baPWV) for diagnosing composite coronary and carotid atherosclerosis has not been completely elucidated.

Method: We enrolled 773 asymptomatic individuals who were referred from 25 public health centers in Seoul and who underwent carotid ultrasonography and coronary computed tomography. Non-invasive hemodynamic parameters, including baPWV, were also measured. Composite coronary and carotid atherosclerosis was defined as follows: 1) coronary artery calcium (CAC) score ≥ 100, 2) coronary artery stenosis (CAS) ≥ 50% of diameter stenosis, 3) carotid intima medial thickness (CIMT) ≥ 0.9 mm, or 4) presence of carotid artery plaque (CAP).

Results: The incidence of composite coronary and carotid atherosclerosis was 28.2%. Coronary atherosclerosis (CAC and CAS) was significantly associated with carotid atherosclerosis (CIMT and CAP). Subjects with higher baPWV (highest quartile) had a higher prevalence of composite coronary and carotid atherosclerosis (p＜.001). Although multivariate analysis failed to show baPWV as an independent predictor for composite atherosclerosis, baPWV had moderate diagnostic power to detect a subject with more than two positive subclinical atherosclerosis exams [area under the curve (AUC), 0.692].

Conclusion: baPWV was associated with the composite coronary and carotid atherosclerotic burden in a community-based asymptomatic population.

Aim: Previous studies have reported that moderate alcohol consumption is protective against cardiovascular disease, but heavy alcohol consumption increases its risk. Endothelial dysfunction is hypothesized to contribute to the development of atherosclerosis and cardiovascular disease. However, few population-based studies have examined a potential effect of alcohol consumption on endothelial function.

Methods: This study included 404 men aged 30–79 years who were recruited from residents in 2 communities under the Circulatory Risk in Communities Study in 2013 and 2014. We asked the individuals about the frequency and volume of alcohol beverages and converted the data into grams of ethanol per day. Endothelial function was assessed by brachial artery flow-mediated dilation (FMD) measurements during reactive hyperemia. We performed cross-sectional analysis of alcohol consumption and %FMD by logistic regression analysis, adjusting for age, baseline brachial artery diameter, body mass index, systolic blood pressure, low-density lipoprotein cholesterol, HbA1c, smoking, antihypertensive medication use, and community.

Results: Individuals who drank ≥ 46 g/day ethanol had a lower age-adjusted mean %FMD than non-drinkers (p＜0.01). Compared with non-drinkers, the age-adjusted odds ratios (ORs) (95% confidence interval) of low %FMD (＜5.3%) for former, light (＜23.0 g/day ethanol), moderate (23.0–45.9 g/day ethanol), and heavy (≥ 46.0 g/day ethanol) drinkers were 1.61 (0.67–3.89), 0.84 (0.43–1.66), 1.09 (0.52–2.25), and 2.99 (1.56–5.70), respectively. The corresponding multivariable-adjusted ORs were 1.76 (0.69–4.50), 0.86 (0.42–1.76), 0.98 (0.45–2.12), and 2.39 (1.15–4.95), respectively.

Conclusions: Heavy alcohol consumption may be an independent risk factor of endothelial dysfunction in Japanese men.

Aim: The efficacy of statin therapy in inducing coronary plaque regression may depend on baseline cholesterol levels. We aimed to determine the efficacy of statin therapy in inducing coronary plaque regression in statin-naïve patients with low cholesterol levels using serial intravascular ultrasound (IVUS) data from the treatment with statin on atheroma regression evaluated by virtual histology IVUS (TRUTH) study.

Methods: The TRUTH study is a prospective, multicenter trial, comparing the efficacies of pitavastatin and pravastatin in coronary plaque regression in 164 patients. All patients were statin-naïve and received statin therapy only after study enrollment. The primary endpoint was the observation of coronary plaque progression, despite statin therapy.

Results: Serial IVUS data, at baseline and after an 8-month follow-up, were available for 119 patients. The patients were divided into three groups based on non-high-density lipoprotein cholesterol (HDL-C) levels—low: ≤140 mg/dl, n=38; moderate: 141-169 mg/dl, n=42; and high: ≥170 mg/dl, n=39. Coronary plaque progression was noted in the low cholesterol group, whereas plaque regression was noted in the moderate and high cholesterol groups [%Δplaque volume: 2.3±7.4 vs. －2.7± 10.7 vs. －3.2±7.5, p=0.004 (analysis of variance)]. After adjusting for all variables, a low non-HDL-C level (≤140 mg/dl) was identified as an independent predictor of coronary plaque progression [odds ratio, 3.7; 95% confidence interval, 1.5-9.1, p=0.004].

Conclusion: Serial IVUS data analysis indicated that statin therapy was less effective in inducing coronary plaque regression in patients with low cholesterol levels but more effective in those with high cholesterol levels at baseline.

University Hospital Medical Information Network (UMIN) (UMIN ID: C000000311).

Aim: Visceral fat accumulation is known to underlie the clustering of cardiovascular risk factors. However, it is not completely understood how visceral fat accumulation influences the development of cardiovascular disease. In this study, we investigated the clinical impact of visceral adiposity on vascular stiffness and thickness in patients with type 2 diabetes (T2D).

Methods: One hundred and sixty-one patients with T2D, including 92 men and 69 women, were included in this cross-sectional study. Visceral fat area (VFA) and subcutaneous fat area (SFA) were measured by dual bioelectrical impedance analysis. Stiffness parameter β and intima-media thickness (IMT) of the common carotid artery were measured by ultrasonography.

Results: The mean age and duration of diabetes in the study population were 61 years and 13.9 years, respectively. In men, VFA and waist circumference (WC) were positively correlated with stiffness parameter β, whereas body mass index (BMI), WC, and SFA were negatively correlated with IMT. In contrast, in women, none of the obesity-related indices were significantly correlated with stiffness parameter β or IMT. In multiple regression analyses, VFA as well as WC, BMI, and SFA were independently associated with stiffness parameter β after adjustment for age and other potential confounders in men but not in women. None of the obesity-related indices were independently associated with IMT for either sex.

Conclusion: In men with T2D, visceral adiposity is associated with carotid arterial stiffness but not thickness.

Aim: Renal insufficiency is associated with worse clinical outcomes in patients with coronary artery disease. Since the introduction of percutaneous coronary intervention (PCI), the revascularization therapy has evolved with advances of devices, improvements in operator techniques, and the establishment of medical therapy. We examined temporal trends of the clinical outcomes following PCI in patients with renal insufficiency.

Methods: Patients with renal insufficiency after PCI at Juntendo University across three eras (plain balloon angioplasty, bare metal stent (BMS), and drug-eluting stent (DES)) were examined in this study. The primary endpoint was a composite of all-cause mortality, nonfatal acute coronary syndrome, nonfatal stroke, and repeat revascularization within 3-years after the index revascularization.

Results: A total of 1,420 patients were examined. Baseline characteristics have become unfavorable over time, whereas administration rate of medications for secondary prevention has increased. The event-free survival rates for the endpoint were different among the groups. Adjusted relative risk reduction for the endpoint was 35% and 51% in the BMS and DES eras (using the plain angioplasty era as reference). The adjusted relative risk reduction of the DES era was 26% compared with that of the BMS era.

Conclusions: The incidence of cardiovascular events after PCI has reduced during the 26-year period mainly because of the reduction in repeat revascularization in patients with renal insufficiency, despite the higher risk profiles in the recent era.

Aim: It is important to explore predictive markers other than conventional cardiovascular risk factors for early detection and treatment of chronic kidney disease (CKD), a major risk factor for end-stage renal failure. We hypothesized that serum albumin and high-sensitivity C-reactive protein (hs-CRP) to be independent markers, and examined their associations with the risk of CKD.

Methods: We examined the associations of serum albumin and hs-CRP levels with the risk of incident CKD, in 2535 Japanese adults aged 40–69 years without CKD at baseline during a median 9.0-year follow-up after adjustment for known cardiovascular risk factors.

Results: During the follow-up period, 367 cases of CKD developed. In multivariable analyses adjusted for known risk factors, the CKD hazard ratios (95% confidence intervals) for the highest versus lowest quartiles of serum albumin levels were 0.69 (0.40–1.17) for men and 0.42 (0.28–0.64) for women. Corresponding values for hs-CRP were 0.95 (0.54–1.67) for men and 1.85 (1.25–2.75) for women. The association of combined serum albumin and hs-CRP with the risk of CKD was examined for women. The hazard ratio was 1.72 (1.17–2.54) for low versus higher albumin levels at lower hs-CRP levels, but such an association was not observed at high hs-CRP level. The hazard ratio was 1.96 (1.44–2.66) for high versus lower hs-CRP levels at higher serum albumin levels, but such association was not observed at low serum albumin level.

Conclusion: Both low serum albumin and high hs-CRP levels were predictive of CKD for women.

Aim: Resveratrol is a popular ingredient in dietary supplements. Some patients concomitantly use dietary supplements and medicines in Japan. In the present study, we determined whether trans-resveratrol and melinjo (Gnetum gnemon L.) seed extract (MSE), which contains resveratrol dimers, interacted with drugs using a mouse model.

Methods: Male C57BL/6J mice were fed experimental diets containing 0.005%, 0.05%, or 0.5% (w/w) trans-resveratrol or MSE for 1 or 12 weeks. The expression of liver cytochrome P-450 (CYP) mRNA and activity of liver microsomal CYP were measured. To determine the influence of resveratrol or MSE on drug efficacy, the anticoagulant activity of warfarin was examined in mice that were fed diets containing trans-resveratrol or MSE for 12 weeks.

Results: When the mice were fed experimental diets for 1 week, none of the doses of trans-resveratrol and MSE affected body weight, liver weight, or plasma AST and ALT levels. Trans-resveratrol also did not affect CYP1A1, CYP1A2, CYP2C, or CYP3A activities. In contrast, 0.5% MSE slightly increased CYP1A1 activity. When the mice were fed experimental diets for 12 weeks, 0.05% trans-resveratrol increased CYP1A1, CYP2C, and CYP3A activities, whereas 0.5% MSE suppressed CYP3A activity. Under these conditions, 0.5% trans-resveratrol enhanced the anticoagulant activity of warfarin, although CYP2C activity increased. However, MSE did not affect the anticoagulant activity of warfarin.

Conclusion: The 0.05% trans-resveratrol did not interact with warfarin in a mouse model, whereas 0.5% trans-resveratrol may have enhanced the anticoagulant activity of warfarin.

Aim: Macrophage apoptosis is a vital event in advanced atherosclerosis, and oxidized low-density lipoprotein (ox-LDL) is a major contributor to this process. Acid sphingomyelinase (ASM) and ceramide are also involved in the induction of apoptosis, particularly in macrophages. Our current study focuses on ASM and investigates its role in ox-LDL-induced macrophage apoptosis.

Methods: Human THP-1 and mouse peritoneal macrophages were cultured in vitro and treated with ox-LDL. ASM activity and ceramide levels were quantified using ultra performance liquid chromatography. Protein and mRNA levels were analyzed using Western blot analysis and quantitative real-time PCR, respectively. Cell apoptosis was determined using Hoechst staining and flow cytometry.

Results: Ox-LDL-induced macrophage apoptosis was triggered by profound endoplasmic reticulum (ER) stress, leading to an upregulation of ASM activity and ceramide levels at an early stage. ASM was inhibited by siRNA or desipramine (DES), and/or ceramide was degraded by recombinant acid ceramidase (AC). These events attenuated the effect of ox-LDL on ER stress. In contrast, recombinant ASM upregulated ceramide and ER stress. ASM siRNA, DES, recombinant AC, and ER stress inhibitor 4-phenylbutyric acid were blocked by elevated levels of C/EBP homologous protein (CHOP); ox-LDL induced elevated levels of CHOP. These events attenuated macrophage apoptosis.

Conclusion: These results indicate that ASM/ceramide signaling pathway is involved in ox-LDL-induced macrophage apoptosis via ER stress pathway.

Aim: The Japan Atherosclerosis Society (JAS) guidelines for the diagnosis and treatment of hyperlipidemia in Japanese adults recommend using low-density lipoprotein cholesterol (LDL-C) calculated by Friedewald formula (F_LDL-C) for subjects with triglyceride (TG) levels ＜400 mg/dL and non-high-density lipoprotein cholesterol (non-HDL-C) levels for subjects with TG levels ≥400 mg/dL. Because small-dense LDL particles are more atherogenic than large LDL particles, we sought the better lipid parameter which was more reflective of the high small-dense LDL-C (sdLDL-C) levels in subjects with TG levels ＜400 mg/dL.

Methods: This study included 769 Japanese subjects who met our inclusion criteria and underwent an annual health examination, including sdLDL-C analyses.

Results: The correlation coefficient of non-HDL-C for sdLDL-C (r=0.760) was significantly higher than that of F_LDL-C (r=0.601). The area under the curve (95% confidence interval) was 0.771 (0.731, 0.811) for F_LDL-C and 0.871 (0.842, 0.901) for non HDL-C, which showed significantly higher predictive value for more than fourth quartile value of sdLDL-C (46 mg/dL). The optimal cut-off point of non-HDL-C was 158 mg/dL. Even in subjects stratified by waist circumstance, homeostasis model assessment of insulin resistance, TG, and F_LDL-C levels and non-HDL-C showed stronger relationships with sdLDL-C than F_LDL-C. Moreover, non-HDL-C showed a better relationship with sdLDL-C than total cholesterol (TC), TC/HDL-C, and non-HDL-C/HDL-C.

Conclusion: Our data suggested that non-HDL-C is superior to F_LDL-C and one of the reliable surrogate lipid markers of sdLDL-C in Japanese subjects with TG levels ＜400 mg/dL.