Plasma Total Cholesterol Research Articles

Tachol1 QTL on mouse chromosome 1 is responsible for hypercholesterolemia and diet-induced obesity.

Hypercholesterolemia raises the risk for cardiovascular complications and overall health. Hypercholesterolemia is common, affecting 10% of the general population of the US, and heritable. Most individuals with hypercholesterolemia have a polygenic predisposition to the condition. Previously we identified a quantitative trait locus, Tachol1, linked to hypercholesterolemia on mouse chromosome 1 (Chr1) in a cross between C57BL/6J (B6) and TALLYHO/JngJ (TH) mice, a polygenic model for human obesity, type 2 diabetes and hyperlipidemia. Subsequently, using congenic mice that carry a TH-derived genomic segment of Chr1 on a B6 background, we demonstrated that the distal segment of Chr1, where Tachol1 maps, is necessary to cause hypercholesterolemia, as well as diet-induced obesity. In this study, we generated overlapping subcongenic lines to the distal segment of congenic region and characterized subcongenic mice carrying the smallest TH region of Tachol1, ~ 16.2Mb in size (B6.TH-Chr1-16.2Mb). Both male and female B6.TH-Chr1-16.2Mb mice showed a significantly increased plasma total cholesterol levels compared to B6 on both chow and high fat (HF) diet. B6.TH-Chr1-16.2Mb mice also had greater fat mass than B6 on HF diet, without increasing food intake. The gene and protein expression levels of absent in melanoma 2 (Aim2) gene were significantly upregulated in B6.TH-Chr1-16.2Mb mice compared to B6. In summary, we confirmed the effect of Tachol1 on hypercholesterolemia and diet-induced obesity using subcongenic analysis.

Pitavastatin ameliorates myocardial injury by inhibiting myocardial inflammation and oxidative stress by modulating the microRNA-106B-5p/mitogen-activated protein kinase kinase kinase 2 axis.

Myocarditis (MC) is a myocardial inflammatory disease that threats human life. Pitavastatin (Pit) is a unique lipophilic statin with potent effects on lowering plasma total cholesterol and triacylglycerols. It has been reported to have pleiotropic effects, such as reducing inflammation and oxidative stress. However, the regulatory mechanism of Pit in MC remains a mystery. Two MC models were established in vitro (lipopolysaccharides-(LPS)-stimulated H9c2 cells) and in vivo (intraperitoneal injection of LPS in mice). The levels of microRNA-106b-5p (miR-106b-5p) and mitogen-activated protein kinase kinase kinase 2 (MAP3K2) were detected. ELISA was used to analyze in vivo cell inflammatory factors and myocardial injury markers, kits were used to detect the expression of antioxidant enzymes, cell counting kit-8 (CCK-8) was used to detect cell proliferation, and flow cytometry was used to detect apoptosis. Hematoxylin and eosin (HE) staining was used to detect the pathological changes of myocardial tissue in mice, and TUNEL staining was used to detect in vivo tissue cell apoptosis. The regulatory mechanism of Pit on miR-106b-5p/MAP3K2 was verified by a series of functional rescue experiments. The results demonstrated that in LPS-induced H9c2 cells, antioxidant enzymes decreased and pro-inflammatory factors and cardiac injury markers increased (p<0.05). However, these phenomenons were attenuated by Pit pretreatment. LPS decreased miR-106b-5p and elevated MAP3K2 in H9c2 cells, while Pit could recover their expression patterns (p<0.05). MAP3K2 was confirmed as a target gene of miR-106b-5p. Upregulating miR-106b-5p or downregulating MAP3K2 could further promote the protective effect of Pit, and vice versa (p<0.05). In addition, in the LPS-induced MC mouse model, histological examination showed that Pit significantly improved the myocardial tissue damage in MC mice, while downregulating miR-106b-5p or upregulating MAP3K2 could suppress the ameliorative effect of Pit (p<0.05). In conclusion, our study demonstrated that Pit ameliorates myocardial injury by suppressing myocardial inflammation and oxidative stress by modulating the miR-106b-5p/MAP3K2 axis.

The impact of chrysanthemi indici flos-enriched flavonoid part on the model of hyperuricemia based on inhibiting synthesis and promoting excretion of uric acid

Ethnopharmacological relevanceIn recent years, in addition to hypertension, hyperglycemia, and hyperlipidemia, the prevalence of hyperuricemia (HUA) has increased considerably. Being the fourth major health risk factor, HUA can affect the kidneys and cardiovascular system. Chrysanthemi Indici Flos is a flavonoid-containing traditional Chinese patent medicine that exhibits a uric acid (UA)-lowering effect. However, the mechanisms underlying Chrysanthemi Indici Flos-enriched flavonoid part (CYM.E) mediated alleviation of HUA remain unelucidated. Aim of the studyThis study aimed to elucidate the efficacy of CYM.E in preventing and treating HUA and its specific effects on UA-related transport proteins, to explore possible mechanism. MethodsThe buddleoside content in CYM.E was determined through high-performance liquid chromatography. HUA was induced in mice models using adenine and potassium oxonate. Subsequently, mice were administered 10 mg/kg allopurinol, and 30, 60, and 90 mg/kg CYM.E to evaluate the effects of CYM.E on the of HUA mice model. Herein, plasma uric acid (UA), creatinine (CR), blood urea nitrogen (BUN), total cholesterol (TC), triglyceride (TG), high-density lipoprotein cholesterol (HDL-c), and low-density lipoprotein cholesterol (LDL-c) contents, along with serum alanine aminotransferase (ALT), and aspartate aminotransferase (AST) activities were measured. Additionally, xanthine oxidase (XOD) and adenosine deaminase (ADA) activities in the liver were determined. The histomorphologies of the liver and kidney tissues were examined through hematoxylin and eosin staining. The messenger RNA (mRNA) expression of facilitated glucose transporter 9 (GLUT9), organic anion transporter (OAT)1, OAT3, and adenosine triphosphate binding cassette subfamily G2 (ABCG2) in the kidney was assessed by real-time quantitative polymerase chain reaction. Furthermore, the expression of urate transporter 1 (URAT1), GLUT9, OAT1, and OAT3 in the kidney, OAT4, and ABCG2 proteins was determined by immunohistochemistry and western blotting. ResultsThe buddleoside content in CYM.E was approximately 32.77%. CYM.E improved body weight and autonomous activity in HUA mice. Additionally, it reduced plasma UA, BUN, and CR levels and serum ALT and AST activities, thus improving hepatic and renal functions, which further reduced the plasma UA content. CYM.E reduced histopathological damage to the kidneys. Furthermore, it lowered plasma TC, TG, and LDL-c levels, thereby improving lipid metabolism disorder. CYM.E administration inhibited hepatic XOD and ADA activities and reduced the mRNA expression of renal GLUT9. CYM.E inhibited the protein expression of renal URAT1, GLUT9, and OAT4, and increased the mRNA and protein expression of renal OAT1, OAT3, and ABCG2. Altogether, these results show that CYM.E could inhibit the production and promote reabsorption of UA and its excretion.

A novel small-molecule PCSK9 inhibitor E28362 ameliorates hyperlipidemia and atherosclerosis.

Proprotein convertase subtilisin/kexin type 9 (PCSK9) binds to the epidermal growth factor precursor homologous domain A (EGF-A) of low-density lipoprotein receptor (LDLR) in the liver and triggers the degradation of LDLR via the lysosomal pathway, consequently leading to an elevation in plasma LDL-C levels. Inhibiting PCSK9 prolongs the lifespan of LDLR and maintains cholesterol homeostasis in the body. Thus, PCSK9 is an innovative pharmacological target for treating hypercholesterolemia and atherosclerosis. In this study, we discovered that E28362 was a novel small-molecule PCSK9 inhibitor by conducting a virtual screening of a library containing 40,000 compounds. E28362 (5, 10, 20 μM) dose-dependently increased the protein levels of LDLR in both total protein and the membrane fractionin both HepG2 and AML12 cells, and enhanced the uptake of DiI-LDL in AML12 cells. MTT assay showed that E28362 up to 80 μM had no obvious toxicity in HepG2, AML12, and HEK293a cells. The effects of E28362 on hyperlipidemia and atherosclerosis were evaluated in three different animal models. In high-fat diet-fed golden hamsters, administration of E28362 (6.7, 20, 60 mg·kg-1·d-1, i.g.) for 4 weeks significantly reduced plasma total cholesterol (TC), triglyceride (TG), low-density lipoprotein-cholesterol (LDL-C) and PCSK9 levels, and reduced liver TC and TG contents. In Western diet-fed ApoE-/- mice (20, 60 mg·kg-1·d-1, i.g.) and human PCSK9 D374Y overexpression mice (60 mg·kg-1·d-1, i.g.), administration of E28362 for 12 weeks significantly decreased plasma LDL-C levels and the area of atherosclerotic lesions in en face aortas and aortic roots. Moreover, E28362 significantly increased the protein expression level of LDLR in the liver. We revealed that E28362 selectively bound to PCSK9 in HepG2 and AML12 cells, blocked the interaction between LDLR and PCSK9, and induced the degradation of PCSK9 through the ubiquitin-proteasome pathway, which finally resulted in increased LDLR protein levels. In conclusion, E28362 can block the interaction between PCSK9 and LDLR, induce the degradation of PCSK9, increase LDLR protein levels, and alleviate hyperlipidemia and atherosclerosis in three distinct animal models, suggesting that E28362 is a promising lead compound for the treatment of hyperlipidemia and atherosclerosis.

Leukemia inhibitory factor receptor inhibition reduces stenosis grade and total serum cholesterol levels in a mouse model for atherosclerosis

Abstract Funding Acknowledgements Type of funding sources: Foundation. Main funding source(s): Dutch Heart Foundation Atherosclerosis is characterized by the accumulation of lipids and immune cells in the arterial wall and it is the main underlying pathology causing cardiovascular diseases (CVD). Cytokines are involved in all stages of atherosclerosis and most cytokines contribute to disease progression. For instance, IL-6 cytokine family members, such as IL-6, oncostatin-M and cardiotrophin-1, are closely related to CVDs. However, the role of leukemia inhibitory factor (LIF), a member of the IL-6 cytokine family, and its receptor (LIFR) in atherosclerosis remains unknown. Therefore, the aim of this study is to assess the contribution of LIFR signaling to atherosclerosis development by systemically inhibiting LIFR in an atherosclerotic mouse model. First, the expression of LIF and LIFR was examined using single-cell RNA sequencing data of human carotid artery plaques. Mast cells highly and exclusively express LIF, whereas LIFR was specifically expressed by activated endothelial cells. To validate LIFR signaling in vivo, female western-type diet fed Ldlr-/- mice were treated with LIFR inhibitor EC359 (5 mg/kg s.c., n=15) or control solvent (n=15) three times per week for eight weeks. During the experiment, weights of the mice did not differ between groups, whereas total serum cholesterol levels were significantly reduced in EC359 treated mice (p=0.03). In particular, EC359 treatment reduced VLDL levels. Additionally, EC359 treatment significantly reduced Sr1, Abca1 and Lrp1 expression in the liver, suggesting an altered lipid metabolism in EC359 treated mice. Furthermore, LIFR inhibition affected innate immunity, both systemically and locally in the plaque. Neutrophil numbers were significantly increased in the circulation (ctrl 18.7±1.7%; EC359 28.0±2.0%; p=0.002) and aortic plaques (ctrl 3.9±0.8%; EC359 9.9±1.9%; p=0.003) of mice treated with EC359 compared to control mice. Moreover, LIFR inhibition affected circulating monocytes resulting in less inflammatory monocytes. The adaptive immune cells, such as T and B cells, remained largely unaffected. Collectively, the reduction of serum total cholesterol and increase in neutrophil numbers in EC359 treated mice resulted in a trend towards reduced atherosclerotic stenosis grade compared to control mice (39.8±1.4% versus 34.9±1.9%; p=0.054), but it did not affect plaque collagen and monocyte/macrophage content in the aortic root. Conclusively, LIFR signaling inhibition reduces atherosclerotic stenosis grade, lowers plasma total cholesterol levels and affects innate immunity.

Comparison of Triglyceride-Glucose Index between Patients with Gallstone and Healthy Individuals.

Triglyceride-glucose index is a product of triglycerides and fasting plasma glucose (FPG) and is a new index of insulin resistance found to correlate with direct measurements. This study aimed to evaluate the relationship between gallstones and triglyceride-glucose index (TGI). A total of 210 patients were included in this retrospective study. Overall, 105 patients with gallstones were included in the patient group. Patients with diabetes mellitus, chronic diseases, malignant diseases, and patients using cholesterol-lowering drugs were excluded from the study. Healthy individuals (105 cases) were selected for the control group. TGI was calculated separately for each individual using the following formula: (TGI) = ln (fasting TG (mg/dL) × fasting glucose (mg/dL)/2). Plasma triglyceride levels were significantly higher in patients with gallstones compared to the control group (P=0.001). Plasma HDL, LDL, and total cholesterol did not differ between the groups (P>0.05). Fasting blood glucose was significantly higher in patients with gallstones compared to the control group (P=0.001). The triglyceride glucose index was significantly higher in patients with gallstones compared to the control group (P<0.001). When the relationship between body mass index and TGI was analyzed, TGI was lower in patients with normal BMI compared to overweight or obese patients (P<0.001). Increased triglyceride/glucose index in patients with gallstones is an indicator of insulin resistance. It is instrumental in demonstrating the presence of insulin resistance in patients with gallstones and may be a useful guide in earlier detection, prevention, and treatment of insulin resistance.

Effects of a Nonlinear Program on Different Health Parameters in the Elderly.

The aim of this research was to evaluate the impact of a nonlinear training program on visceral adipose tissue (VAT) and systolic (SBP) and diastolic (DBP) blood pressure, as well as the response of biochemical parameters such as fasting plasma glucose (FPG), total cholesterol (TC), high-density cholesterol (HDL-C), low-density cholesterol (LDL-C), and triglycerides (TG). The nonlinear periodized program would produce greater improvements in outcomes than the linear periodized training program. Randomized cross-sectional design. Level 3. Older adults with no previous training experience (10 male and 8 female [age, 64 ± 2.1 years; height, 165.12 ± 7.5 cm; body mass, 72.5 ± 11.4 kg; body max index, 26.5 ± 3.2 kg/m2]) were randomized to linear (n = 9, TT) or undulating (n = 9, UT) periodization. After a 3-week familiarization period, all participants performed 3 sessions of resistance training per week; 8 weeks of training were conducted for each group. Dual x-ray absorptiometry was used to measure VAT, and SBP and DBP were measured using an OMRON M3 digital automatic blood pressure monitor. Blood samples were collected between 8:00 a.m. and 9:30 a.m. after 12-hour overnight fasting. Both interventions significantly (P < 0.05) decreased FPG, TC, LDL-C, and TG. A significant decrease in SBP and DBP was observed only in the UT group (P < 0.05). No significant between-group differences in outcomes were observed (P > 0.5). However, the effect size was marginally more pronounced for all outcomes in the UT group. An undulating periodization program was effective in improving VAT, TC, LDL-C, FPG, HDL-C, TG, and blood glucose levels in older adults. Resistance training can be programmed in an undulating or traditional way in older adults based on improvements in health parameters, considering adherence and individual preferences.

Low Plasma Lipids Are Associated with Relapsing and Lethal Visceral Leishmaniasis in HIV-Infected Patients.

Visceral leishmaniasis (VL) results from protozoa Leishmania infantum and L. donovani infection. This study investigated whether host factors would explain the relapses. First, susceptibility to amphotericin B of L. infantum isolates was evaluated in vitro. Then, clinical data and the lipid profile of patients with relapsing and non-relapsing VL were assessed. Susceptibility to amphotericin B was similar between the isolates. CD4+ lymphocytes were reduced in both groups of patients in the first episode and with relapsing VL. Still, the strongest blood cell indicator associated with relapses was low total lymphocyte counts. Total plasma cholesterol, high-density lipoprotein, low-density lipoprotein, and, uniquely, triglycerides of the six individuals in the first episode and twenty-three with relapsing VL were lower in relapsing patients than those in the first episode. Deceased patients had extremely low low-density lipoprotein. After CD4+ decreases, lymphocyte CD8+ reduction is the final stage of immunological failure. The lower lipid concentrations appear to be secondary to the depletion of fat stores by inflammation-induced cachexia and fat exhaustion provoked by the co-occurrence of both diseases, which can finally lead to death.

Effect of virgin olive oil as spreadable preparation on atherosclerosis compared to dairy butter in Apoe-deficient mice

Olive oil is the main source of lipid energy in the Mediterranean diet and there is strong evidence of its health benefits. The effect of extra virgin olive oil (EVOO) in the form of a preparation of spreadable virgin olive oil (S-VO) on the progression of atheroma plaques was investigated in Apoe-deficient mice, a model of accelerated atherosclerosis. Methods: Two isocaloric Western purified diets containing 20% fat, either as S-VO or as dairy butter, were used to feed 28 males and 16 females of two-month-old Apoe-deficient mice for 12 weeks. S-VO was prepared by blending more than 75% virgin olive oil with other vegetal natural fat to obtain a solid fat. Plasma total cholesterol, triglycerides and HDL cholesterol were measured. Hepatic lipid droplets were analyzed. Areas of atherosclerotic aortic lesions were quantified in cross-sectional images of the proximal aorta and en face analysis of the whole aorta. Results: Total plasma cholesterol was increased in mice on the butter-supplemented diet in both female and male mice compared to S-VO, and the ratio of TC/HDL-cholesterol was significantly lower in S-VO than in the butter diet, although only in males, and no differences in plasma triglycerides were observed. No significant differences in hepatic lipid droplets were observed between diets in either sex. Aortic lesion areas were significantly higher in mice consuming the butter versus the S-VO diet in both sexes. Conclusion: Extra virgin olive oil prepared in spreadable form maintained the delay in atheroma plaque progression compared to butter.

Candida albicans accelerates atherosclerosis by activating intestinal hypoxia-inducible factor2α signaling

The gut microbiota is closely linked to atherosclerosis. However, the role of intestinal fungi, essential members of the complex microbial community, in atherosclerosis is poorly understood. Herein, we show that gut fungi dysbiosis is implicated in patients with dyslipidemia, characterized by higher levels of Candida albicans (C.albicans), which are positively correlated with plasma total cholesterol and low-density lipoprotein-cholesterol (LDL-C) levels. Furthermore, C.albicans colonization aggravates atherosclerosis progression in a mouse model of the disease. Through gain- and loss-of-function studies, we show that an intestinal hypoxia-inducible factor 2α (HIF-2α)-ceramide pathway mediates the effect of C.albicans. Mechanistically, formyl-methionine, a metabolite of C.albicans, activates intestinal HIF-2α signaling, which drives increased ceramide synthesis to accelerate atherosclerosis. Administration of the HIF-2α selective antagonist PT2385 alleviates atherosclerosis in mice by reducing ceramide levels. Our findings identify a role for intestinal fungi in atherosclerosis progression and highlight the intestinal HIF-2α-ceramide pathway as a target for atherosclerosis treatment.

The association of body mass index variability with cardiovascular disease and mortality: a mediation analysis of pooled cohorts.

We aimed to investigate the effect of BMI variability on CVD and mortality and to explore the mediation effects of the main cardiovascular risk factors contributing to this association. Participants aged 40-65 years were pooled from three cohort studies(ARIC [Atherosclerosis Risk in Communities], MESA [Multi-ethnic Study of Atherosclerosis], and TLGS [Tehran Lipid and Glucose Study]. We employed root mean squared error of the fractional mixed model to calculate BMI variability in the measurement period. In the event assessment period, the hazard ratios for CVD and mortality were estimated using Cox proportional hazard regression models. In the next step, the mediation and interaction effects of fasting plasma glucose, total cholesterol, and systolic blood pressure were determined. A total of 19073 participants were included in this pooled analysis. During a median of 20.7 years of follow-up, 3900 (20.44%) CVD and 6480 (33.97%) all-cause mortality events were recorded. After adjusting for potential confounders, BMI variability was linked to the 1.3 (1.2-1.4) and 1.7 (1.6-1.8) increased risk of CVD and mortality, respectively. Fasting plasma glucose mediated approximately 24% and 8% of the effect of BMI variability on CVD and mortality, respectively. However, systolic blood pressure and total cholesterol did not have mediation effects in this association. High BMI variability is independently associated with the development of CVD and mortality. This association is partly mediated through fasting plasma glucose. Modern cardiometabolic therapies that lower fasting glucose may reduce the risk of future CVD and mortality in individuals with high BMI variability.

Weight loss, glycolipid profile changes in type 2 diabetes patients after esophagectomy: a propensity score matching analysis

BackgroundType 2 diabetes mellitus (T2DM) is a common co-morbidity in patients who receive esophagectomy and has unfavorable effects on glucose and lipid metabolism in patients. This study examines how weight and glycolipid metabolism change in patients with T2DM following esophagectomy.MethodsThis retrospective, one-center, observational analysis with a propensity score matching analysis (PSM) included 114 patients who underwent esophageal surgery in the Department of Cardiothoracic Surgery, the 900th Hospital of Joint Logistic Support Force from 2017 to 2020, which were separated into T2DM group and Non-T2DM group. Weight, body mass index (BMI), fasting plasma glucose (FPG), triglycerides (TG), total cholesterol (TC), high-density lipoprotein (HDL), and low-density lipoprotein (LDL) were measured and analyzed before and after the operation.ResultsTwo groups showed similar reductions in weight and BMI after surgery. In the T2DM group, weight decreased from 63.10(10.31) before surgery to 55.10(11.60) kg at 6 months (P < 0.001) with BMI decreasing from 22.67 (2.90) to 19.77 (3.48); While in the Non-T2DM group, weight decreased from 61.42 (8.46) to 53.19 (9.26) kg at 6 months after surgery with BMI decline from 22.49 (2.77) before operation to 19.45 (3.08) at 6 months after surgery. Fasting plasma glucose levels showed a significant decrease (P = 0.035) in the T2DM group at a six-month point of 7.00 (2.21) mmol/L compared to preoperative levels of 7.67 (2.32) mmol/L. HDL levels increased significantly in the Non-T2DM group at six months postoperatively at 1.52 (0.05) with P < 0.001 compared to preoperative levels of 1.22(0.04) mmol/L. TG, LDL, and TC levels decreased significantly in both groups from the preoperative to the 6-month point.ConclusionsEsophagectomy induces weight loss in T2DM and Non-T2DM groups, improves long-term glucose metabolism in the T2DM group, and enhances lipid metabolism in both groups. Further research is needed to understand their mechanisms.

Use of Bacillus subtilis multiplicate in the water used for biofloc formation: Growth, hemato-biochemistry, intestinal bacteria colonies, and bacterial resistance evaluations of Nile tilapia

Probiotics in the biofloc technology (BFT) system improve fish growth and immune response and promote a favorable intestinal microbiota balance. This study aimed to investigate the growth, hemato-biochemical parameters, bacterial resistance, and the number of intestinal bacteria colonies of Nile tilapia (6.23 g) reared in the BFT system with graded levels of Bacillus subtilis multiplicate (BSM) in the water. The multiplication of these microorganisms was performed in a biofactory. Fish were distributed into four different groups with graded BSM concentrations in the water of 0.00, 0.25, 0.50, and 0.75% (or 0; 750; 1500; and 2250 mL; called BSM0 (control), BSM25, BSM50, and BSM75, respectively). The experimental procedure had 45 days for growth performance evaluations and seven additional days of bacterial challenge with Aeromonas hydrophila. The experimental design was randomized into four treatments in triplicate (n = 20 fish per tank; initial stocking density of 0.50 kg m−3). Survival was 100% before and after the bacterial challenge. Increasing BSM in the BFT system had a positive linear effect on nitrite, nitrate, settled solids, hemoglobin, and hematimetric indices. BSM50 showed the highest specific growth rate, weight gain, intestinal total bacteria, and B. subtilis counts and the lowest feed conversion rate. In BSM50 and BSM75, liver glycogen levels and aspartate aminotransferase (AST) activity were higher and lower, respectively, than in the other treatments. Plasma total cholesterol, total proteins, albumin, and skin mucus total proteins and albumin were highest in BSM25. After the bacterial challenge, the number of lesions was lower in BSM75 than in the control group. In conclusion, we recommended BSM50 because it increased the growth performance and bacterial colonization in the intestine. It also decreased liver AST activity in Nile tilapia reared in the BFT system without compromising water quality and hematological and plasmatic parameters.

Inhibition of activin receptor 2 signalling ameliorates metabolic dysfunction–associated steatotic liver disease in western diet/L-NAME induced cardiometabolic disease

ObjectiveBlockade of activin 2 receptor (ACVR2) signaling has been shown to improve insulin sensitivity and aid in weight loss. Inhibition of ACVR2 signaling restores cardiac function in multiple heart failure models. However, its potential in the treatment of obesity-related cardiometabolic disease remains unknown. Here, we investigated targeting ACVR2 signaling in cardiometabolic disease manifested with metabolic dysfunction–associated steatotic liver disease (MASLD). MethodsMice were fed a high-fat, high-sugar diet combined with the administration of nitric oxide synthase inhibitor L-NAME in drinking water, which causes hypertensive stress. For the last eight weeks, the mice were treated with the soluble ACVR2B decoy receptor (sACVR2B-Fc). ResultssACVR2B-Fc protected against the development of comorbidities associated with cardiometabolic disease. This was most pronounced in the liver where ACVR2 blockade attenuated the development of MASLD including cessation of pro-fibrotic activation. It also significantly reduced total plasma cholesterol levels, impeded brown adipose tissue whitening, and improved cardiac diastolic function. In vitro, ACVR2 ligands activin A, activin B and GDF11 induced profibrotic signaling and the proliferation of human cardiac fibroblasts. ConclusionsBlockade of ACVR2B exerts broad beneficial effects for therapy of cardiometabolic disease. By reducing obesity, ameliorating cardiovascular deterioration and restraining MASLD, blockade of ACVR2B signaling proves a potential target in MASLD and its comorbidities.

Abstract 1005: Hypertriglyceridemia Promotes Aortic Aneurysm Formation And Rupture In Angiotensin Ii Infused Mice

Plasma total cholesterol (TC) and triglyceride (TG) are reported to be positively associated with the risk of abdominal aortic aneurysm (AAA). Our group discovered that hypercholesterolemia augments angiotensin II-induced AAA in mice. We also found that remnants of triglyceride-rich lipoproteins (TRLs) may be crucial for AngII-induced AAA. Recently, it was reported that hypertriglyceridemia (HTG) caused by inducible lipoprotein lipase deficiency (iLpl-/-) results in uptake and accumulation of TRL lipid in aortic endothelial cells in vivo. Based upon these findings, we hypothesized that HTG stimulates AAA formation. Methods: Adult male and female Lplf/f.beta-actin-Mer/Cre/Mer 1/0 and Lplf/f mice were administered 75 mg/kg/day tamoxifen for 5 consecutive days. Mice were fed either a standard diet (SD) throughout the study or a Western-type diet (WD) starting 1 wk after the completion of tamoxifen administration and continuing for a total of 5 wks. Mini osmotic pumps were implanted in mice 2 wks after completion of tamoxifen administration and delivered saline or AngII at 1,000 ng/kg/min for 4 wks. Results: AngII infused and SD fed iLpl-/- versus Lplf/f mice had elevated plasma TG and TC levels but similar abdominal aortic external diameter and AAA incidence. Since the SD was low in fat, a high fat WD was then fed to AngII-infused mice. Plasma TG and TC were increased in female iLpl-/- versus Lplf/f mice. However, plasma lipid concentrations could not be measured in male iLpl-/- because 10 of 11 animals had died from aortic rupture. In contrast, none of the male Lplf/f mice died and only 4 of 11 had abdominal aortic dilation. Female compared to male iLpl-/- mice were protected from AngII-induced aortic rupture (1/11 died) and only 1 of 9 females had abdominal aortic dilation. To eliminate the possibility that HTG alone caused aortic rupture in male mice, the study was repeated with the addition of saline infusion. In agreement with the first study, AngII plus WD lead to death by aortic rupture in all male iLpl-/- mice (9/9). In contrast, iLpl-/- male mice infused with saline had markedly greater survival (1/5 died of unknown cause). Conclusions: HTG causes aortic aneurysm development in AngII-infused male iLpl-/- mice. Thus, treating HTG could reduce AAA risk.

Abstract 2067: Myeloid Cell Lgr4 Contributes To Atherosclerotic Lesion Formation

Background: R-Spondin 2 (RSPO2), a matricellular protein, primarily functions via binding to cell surface receptors including leucine-rich repeat-containing G protein-coupled receptor (LGR) 4, 5, and 6. We previously identified elevated RSPO2 levels in both human and mouse atherosclerotic arteries. Our preliminary experiments demonstrated increased expression of LGR4 in human atherosclerotic aortic segments, identified Lgr4 as the major RSPO2’s receptor in primary murine macrophages, and atherogenic molecule oxLDL stimulated LGR4 levels in macrophages. However, the precise role of macrophage LGR4 in atherosclerosis development is unknown. Methods and Results: We generated novel myeloid cell-specific Lgr4 knockout mice ( Lgr4 fl/fl LysM Cre +/- , Lgr4 ΔM ) by breeding Lgr4 fl/fl mice with LysM Cre +/- mice. AAV8-h PCSK9 -injected Lgr4 ΔM and littermate control Lgr4 fl/fl ( Lgr4 WT ) were fed a Western diet (16 weeks) to investigate for atherosclerosis. Oil red O (ORO) staining of whole aortas (en face) and aortic root sections showed reduced atherosclerosis in male Lgr4 ΔM mice compared with sex-matched Lgr4 WT mice. Further histochemical staining performed on aortic root sections revealed smaller necrotic cores in Lgr4 ΔM mice. However, there were no significant differences in plasma total cholesterol and body weight. Interestingly, male Lgr4 ΔM mice had decreased fat mass percentage, blood glucose, and epididymal adipose tissue weight. Initial studies with female mice ( n = 4/group) demonstrated no differences in atherosclerosis and metabolic parameters. Additional in vitro studies exhibited that RSPO2 treatment induces oxLDL uptake, reduces efferocytic capacity, and promotes activation of VASP protein (inducer of actin aggregates) in macrophages. Further, the deletion of Lgr4 reduces lipid accumulation in macrophages. Conclusions: Taken together, these findings suggest that deletion of myeloid cell Lgr4 suppresses atherosclerotic lesion formation and identify Lgr4 as a novel therapeutic target for atherosclerosis.

Abstract 3057: Vascular Smooth Muscle Cell Cd47 Contributes To Atherosclerosis Development

Aims: Thrombospondin-1 (TSP1), a secretory protein, has been associated with various cardiovascular diseases. TSP1 mainly functions via its cognate receptors CD47 and CD36. Recently, we observed elevated TSP1 expression in human and murine atherosclerotic arteries. However, the role of vascular smooth muscle cell (VSMC) TSP1-CD47 signaling in regulating VSMC phenotype and atherogenesis is unclear. Methods and Results: Human arterial SMCs were treated with human TSP1 and analyzed for SMC markers and proliferation. The data revealed decreased expression of SMC markers, ACTA, CNN1, and SM22α in TSP1-exposed VSMCs compared with vehicle-treated control cells. Further, TSP1 treatment increased VSMC proliferation in vitro , suggesting TSP1 induces VSMC hyperproliferative phenotype. Additional experiments showed CD47 as the major TSP1’s receptor in VSMCs. To investigate the in vivo role of VSMC Cd47 in atherosclerosis, we generated tamoxifen-inducible SMC-specific Cd47 knockout mice ( Cd47 f/f Myh11 Cre +/– , Cd47 ΔVSMC ) by crossing female Cd47 f/f mice with male Myh11 Cre +/- mice. Male tamoxifen-injected Cd47 ΔVSMC and corn oil-administered Cd47 WT mice were given a single injection of AAV8- hPCSK9 (1х10 11 viral genomes/mouse, IP) and fed a Western diet for 12 weeks. En face oil red O (ORO) staining of whole aortas exhibited reduced atherosclerosis in Cd47 ΔVSMC mice compared with control Cd47 WT mice. Further histochemical staining performed on aortic root sections revealed attenuated neointimal lesion area, lipid accumulation and necrotic area in Cd47 ΔVSMC mice. However, there were no significant differences in weight gain, body composition (fat and lean mass), plasma total cholesterol, and fasting blood glucose between Cd47 ΔVSMC and Cd47 WT mice. Conclusions: Taken together, these findings suggest that TSP1 promotes VSMC phenotype switch and VSMC-specific Cd47 deletion in hypercholesterolemic mice suppresses atherosclerosis progression.

Exploring the relationship between dietary rapeseed meal, condensed tannin and growth, nutrive metabolism in largemouth bass (Micropterus salmoides)

This study evaluated the effects of rapeseed meal (RM) and condensed tannin (CT) on the growth and nutritional metabolism of largemouth bass (Micropterus salmoides), and speculated whether there was interaction between anti-nutritional factors in RM. Eight diets were formulated for 55-day feeding trial, six of which were practical diet groups, including control group (CG), five RM inclusive groups with graded RM levels of 5% (RM5), 10% (RM10), 15% (RM15), 20% (RM20) and 25% (RM25) partially replacing fishmeal, and the other two were semi-purified diets with 0.062% CT (CT5) ≈ the CT content in the RM5 and 0.31% CT (CT25) ≈ the CT content in the RM25. The results of the experiments on replacing fishmeal with RM demonstrated that with the increase of RM substitution level, the weight gain rate (WGR), survival rate (SR) and specific growth rate (SGR) of largemouth bass showed a downward trend. High levels of RM led to the inhibition of digestive enzyme activity in largemouth bass, as well as the manifestation of abnormal markers related to protein, fat, and carbohydrate metabolism in both plasma and liver. RM reduced muscle protein and fat content, although the effect was not statistically significant at 5% RM level. The results of CT concentration experiments showed that the addition of 0.31% CT significantly reduced WGR, SGR, SR, protein efficiency ratio (PER) and gastrointestinal digestive enzyme activity, and significantly increased the feed coefficient ratio (FCR), plasma ammonia (PA), plasma glucose (PG), and hepatic glycogen (HG) and liver triglyceride (TG). The addition of 0.062% CT significantly reduced PG and HG. Supplementation of CT significantly reduced muscle fat. Among the groups of CT supplementation and RM, CT5 exhibited superior WGR, PER and FCR compared to RM5, along with enhanced intestinal trypsin, amylase activity, liver glutamic pyruvic transaminase (GPT) activity and decreased content of HG and liver TG; CT25 exhibited decreased activities of gastric lipase (LPS), enteric trypsin and pepsin, and increased content of total cholesterol (T-CHO) and glucose in plasma in compariosn to RM25, accompanied with no significant change in growth performance. Therefore, largemouth bass exhibited tolerance to a 5% inclusion of RM in the feed without compromising their growth, digestion and metabolic functions. The incorporation of 5% RM, equivalent to a supplemental 0.062% CT, had regulatory effects on the digestion and metabolism of largemouth bass while improving feed utilization. There was an interaction among anti-nutritional factors in RM, with synergistic effect being evident at low RM levels, whereas antagonism more promient at high levels of RM.

Correction of plasma fat-soluble vitamin levels by blood lipids in elderly patients with coronary heart disease

This study aims to investigate the correlation between plasma fat-soluble vitamin levels and blood lipid in elderly patients with coronary heart disease (CHD). A total of 120 participants were enrolled, including 60 CHD patients and 60 controls without CHD. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) was used to quantify plasma levels of vitamins A, D3, E, and K. Data analysis was conducted using the statistical analysis system module of MetaboAnalyst 5.0. The CHD group showed significantly higher levels of plasma total cholesterol (TC), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) but not high-density lipoprotein cholesterol (HDL-C) compared to controls. The CHD group exhibited significantly higher plasma levels of VA and VE, positively correlating with TC, TG, and LDL-C. After adjusted by TG levels, the CHD group had significantly lower plasma levels of VA and VE, negatively correlating with TC, TG, and LDL-C. The CHD group also had significantly lower concentrations of VD3, independent of TG modification, compared to controls. VD3 negatively correlated with TC, TG, and LDL-C. Elderly individuals with CHD display abnormal blood lipid metabolism, and fat-soluble vitamins adjusted by TG levels can more accurately and timely response to implicit fat-soluble vitamins deficiency in CHD patients.

A long-acting LEAP2 analog reduces hepatic steatosis and inflammation and causes marked weight loss in mice

ObjectiveThe number of individuals affected by metabolic dysfunction associated fatty liver disease [1] is on the rise, yet hormonal contributors to the condition remain incompletely described and only a single FDA-approved treatment is available. Some studies suggest that the hormones ghrelin and LEAP2, which act as agonist and antagonist/inverse agonist, respectively, for the G protein coupled receptor GHSR, may influence the development of MAFLD. For instance, ghrelin increases hepatic fat whereas synthetic GHSR antagonists do the opposite. Also, hepatic steatosis is less prominent in standard chow-fed ghrelin-KO mice but more prominent in 42% high-fat diet-fed female LEAP2-KO mice. MethodsHere, we sought to determine the therapeutic potential of a long-acting LEAP2 analog (LA-LEAP2) to treat MAFLD in mice. LEAP2-KO and wild-type littermate mice were fed a Gubra-Amylin-NASH (GAN) diet for 10 or 40 wks, with some randomized to an additional 28 or 10 days of GAN diet, respectively, while treated with LA-LEAP2 vs Vehicle. Various metabolic parameters were followed and biochemical and histological assessments of MAFLD were made. ResultsAmong the most notable metabolic effects, daily LA-LEAP2 administration to both LEAP2-KO and wild-type littermates during the final 4 wks of a 14 wk-long GAN diet challenge markedly reduced liver weight, hepatic triglycerides, plasma ALT, hepatic microvesicular steatosis, hepatic lobular inflammation, NASH activity scores, and prevalence of higher-grade fibrosis. These changes were accompanied by prominent reductions in body weight, without effects on food intake, and reduced plasma total cholesterol. Daily LA-LEAP2 administration during the final 10 d of a 41.5 wk-long GAN diet challenge also reduced body weight, plasma ALT, and plasma total cholesterol in LEAP2-KO and wild-type littermates and prevalence of higher grade fibrosis in LEAP2-KO mice. ConclusionsAdministration of LA-LEAP2 to mice fed a MAFLD-prone diet markedly improves several facets of MAFLD, including hepatic steatosis, hepatic lobular inflammation, higher-grade hepatic fibrosis, and transaminitis. These changes are accompanied by prominent reductions in body weight and lowered plasma total cholesterol. Taken together, these data suggest that LEAP2 analogs such as LA-LEAP2 hold promise for the treatment of MAFLD and obesity.