ABSTRACT
Osteoprotegerin (OPG) is a soluble decoy receptor for receptor activator of nuclear factor kB ligand (RANKL), and is implicated in the pathogenesis of atherosclerosis. The aim of the present study was to examine the hypothesis that serum OPG concentrations are increased in patients with stable coronary artery disease (CAD) at different serum levels of soluble RANKL (sRANKL). The study used a case-control design in which consecutively hospitalized individuals were recruited. Fasting blood samples were taken upon admission for serum testing. Participants with previously diagnosed CAD that was asymptomatic or had controlled symptoms constituted the stable CAD group, whereas patients with negative coronary computed tomography angiography results constituted the control non-CAD group. Exclusion criteria included recent acute coronary syndrome, severe heart failure, CAD-complicating autoimmune, blood or thyroid diseases, cancer, elevated temperature with or without infection, severe liver or kidney dysfunction, abnormal calcium metabolism, recent surgery and trauma history. A total of 118 individuals were included in the study. Smoothed plots generated using the recursive method and multivariate models showed that the incidence of stable CAD increased with serum OPG level up to the turning point of 18 pg/ml. This trend was observed at both high [odds ratio (OR), 1.61; 95% confidence interval (CI), 1.04-2.50; P=0.032) and low sRANKL concentrations (OR, 1.52; 95% CI, 1.06-2.17; P=0.022) after adjustment for cardiovascular risk factors. In conclusion, serum OPG levels ≤18 pg/ml are positively associated with stable CAD, regardless of sRANKL levels. In addition, at the same serum OPG level, higher sRANKL levels are associated with a greater incidence of stable CAD compared with lower sRANKL levels. This study identified the relationship between OPG, sRANKL, and stable CAD, and established the reference range for future clinical use.
ABSTRACT
In this study, the hypothesis that Wnt/ß-catenin pathway is involved in the arterial calcification by regulating the osteoprotegerin (OPG)/receptor activator of NF-κB ligand (RANKL) system was tested. The ß-catenin expression was measured in the warfarin-induced calcified arteries and the osteoblast-like cells differentiating from smooth muscle cells (SMCs) by immunohistochemistry and Western blotting. The Wnt/ß-catenin pathway was activated or inhibited by lithium chloride (LiCl) or dickkopf 1 (DKK1) in vitro and in vivo. Then the calcification level was determined by von Kossa staining, Ca2+ content assay, and alkaline phosphatase (ALP) activity assay. The expression levels of osteocalcin, OPG and RANKL were detected by Western blotting or real-time PCR. The results showed that in calcified arteries and OBL cells, the activation of Wnt/ß-catenin pathway significantly enhanced the calcification as evidenced by increased von Kossa stains, Ca2+ contents, ALP activities, and osteocalcin expression levels (P<0.05), and it promoted the RANKL expression (P<0.05), but slightly affected the OPG expression. These results indicated that the activation of Wnt/ß-catenin pathway worsens the arterial calcification, probably by promoting the RANKL expression.
Subject(s)
Muscle, Smooth, Vascular/blood supply , Osteoprotegerin/genetics , RANK Ligand/genetics , Vascular Calcification/metabolism , Warfarin/adverse effects , Wnt Signaling Pathway/drug effects , Animals , Calcium/metabolism , Cell Differentiation , Cells, Cultured , Disease Models, Animal , Gene Expression Regulation/drug effects , Humans , Intercellular Signaling Peptides and Proteins/pharmacology , Lithium Chloride/pharmacology , Male , Muscle, Smooth, Vascular/cytology , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/metabolism , Osteoprotegerin/metabolism , RANK Ligand/metabolism , Rats , Vascular Calcification/chemically induced , Vascular Calcification/genetics , beta Catenin/metabolismABSTRACT
The initiation and progression of atherosclerotic cardiovascular disease (ASCVD) has always been associated with a series of risk factors. Evidences of statin therapy from randomized clinical trials are abundant, whereas discussions regarding patients with ASCVD without evidence-based risk factors are rare. Here, we describe a case of a 58-year-old woman who was diagnosed with ASCVD with none of these evidence-based risk factors. After four years of medical interventions, including atorvastatin, the patient recovered completely from severe chest pain with significant regression of atherosclerotic plaques in coronary arteries.
Subject(s)
Cardiovascular Diseases/drug therapy , Coronary Artery Disease/drug therapy , Plaque, Atherosclerotic/drug therapy , Atorvastatin/administration & dosage , Atorvastatin/therapeutic use , Cardiovascular Diseases/pathology , Computed Tomography Angiography/methods , Coronary Artery Disease/diagnosis , Coronary Vessels/diagnostic imaging , Coronary Vessels/pathology , Female , Humans , Hydroxymethylglutaryl-CoA Reductase Inhibitors/therapeutic use , Middle Aged , Randomized Controlled Trials as Topic , Risk Factors , Treatment OutcomeABSTRACT
Heart failure (HF) is the end stage of various kinds of cardiovascular diseases and leads to a high mortality worldwide. Numerous studies have demonstrated that frequencies of CD4+CD25+Foxp3+ regulatory T cells (Tregs) are reduced in HF patients and properly expanding Tregs attenuates HF progression. Histone deacetylase (HDAC) 9 has been revealed to contribute to several cardiovascular and cerebrovascular diseases. Plenty of studies showed that HDAC9 negatively regulated the number and function of Tregs. Thus, we aim to investigate the expression of HDAC 9 in patients with chronic heart failure (CHF) and the relationship among HDAC9, Tregs and CHF. Our research showed a reduced number of Tregs and an increased expression of HDAC9 mRNA in CHF patients. Patients with CHF were divided into two groups by heart function grade of New York Heart Association (NYHA), we found that the HDAC9 mRNA expression level in NYHA grade II-III group were lower than that in NYHA grade IV group. More importantly, the correlation study suggested that the expression of HDAC9 mRNA was negatively correlated to Tregs frequency and left ventricular ejection fraction (LVEF), whereas positively correlated to larger left ventricular end-diastolic dimension (LVEDD) and B-type natriuretic peptide (BNP) in patients with CHF. The correlation studies also showed a positive correlation between HDAC9 and the severity of CHF. Our research suggests that HDAC9 may be a new indicator for assessing CHF and it may offer a new direction for research of CHF.
Subject(s)
Heart Failure/enzymology , Heart Failure/immunology , Histone Deacetylases/metabolism , Repressor Proteins/metabolism , T-Lymphocytes, Regulatory/enzymology , Aged , Chronic Disease , Female , Forkhead Transcription Factors/genetics , Forkhead Transcription Factors/metabolism , Heart Failure/genetics , Heart Failure/physiopathology , Heart Function Tests , Histone Deacetylases/genetics , Humans , Lymphocyte Count , Male , Middle Aged , RNA, Messenger/genetics , RNA, Messenger/metabolism , Repressor Proteins/geneticsABSTRACT
Coronary artery disease (CAD) is a multifactorial disease in which inflammation plays a central role. This study aimed to investigate the association of inflammatory markers such as the neutrophil to lymphocyte ratio (NLR), the Global Registry of Acute Coronary Events (GRACE) score with in-hospital mortality of elderly patients with acute myocardial infarction (AMI) in an attempt to explore the prognostic value of these indices for elderly AMI patients. One thousand consecutive CAD patients were divided into two groups based on age 60. The laboratory and clinical characteristics were assessed retrospectively by reviewing the medical records. The NLR and GRACE score were calculated. In the elderly (≥60 years), patients with non-ST-elevation myocardial infarction (NSTEMI) and ST-elevation myocardial infarction (STEMI) had significantly higher NLR than did those with unstable angina (UA) and stable angina pectoris (SAP) (P<0.01). The NLR was considerably elevated in older AMI patients compared with their younger counterparts (<60 years) (P<0.05). In elderly AMI patients, the NLR was considerably higher in the high-risk group than in both the low-risk and medium-risk groups based on the GRACE score (P<0.05 and P<0.01, respectively), and the NLR was positively correlated with the GRACE score (r=0.322, P<0.001). Either the NLR level or the GRACE score was significantly higher in the death group than in the surviving group (P<0.05). By curve receiver operator characteristic curve (ROC) analysis, the optimal cut-off levels of 9.41 for NLR and 174 for GRACE score predicted in-hospital death [ROC area under the curve (AUC) 0.771 and 0.787, respectively, P<0.001]. It was concluded that an elevated NLR is a potential predictor of in-hospital mortality in elderly patients with AMI.
Subject(s)
Hospital Mortality , Lymphocytes/pathology , Myocardial Infarction/blood , Myocardial Infarction/mortality , Neutrophils/pathology , Aged , Aged, 80 and over , Biomarkers/blood , Female , Humans , Inflammation/blood , Lymphocyte Count , Male , Myocardial Infarction/immunology , Prognosis , ROC Curve , Survival AnalysisABSTRACT
BACKGROUND/AIMS: Baicalin has been shown to be effective for various animal models of cardiovascular diseases, such as pulmonary hypertension, atherosclerosis and myocardial ischaemic injury. However, whether baicalin plays a role in cardiac hypertrophy remains unknown. Here we investigated the protective effects of baicalin on cardiac hypertrophy induced by pressure overload and explored the potential mechanisms involved. METHODS: C57BL/6J-mice were treated with baicalin or vehicle following transverse aortic constriction or Sham surgery for up to 8 weeks, and at different time points, cardiac function and heart size measurement and histological and biochemical examination were performed. RESULTS: Mice under pressure overload exhibited cardiac dysfunction, high mortality, myocardial hypertrophy, increased apoptosis and fibrosis markers, and suppressed cardiac expression of PPARα and PPARß/δ. However, oral administration of baicalin improved cardiac dysfunction, decreased mortality, and attenuated histological and biochemical changes described above. These protective effects of baicalin were associated with reduced heart and cardiomyocyte size, lower fetal genes expression, attenuated cardiac fibrosis, lower expression of profibrotic markers, and decreased apoptosis signals in heart tissue. Moreover, we found that baicalin induced PPARα and PPARß/δ expression in vivo and in vitro. Subsequent experiments demonstrated that long-term baicalin treatment presented no obvious cardiac lipotoxicity. CONCLUSIONS: The present results demonstrated that baicalin attenuates pressure overload induced cardiac dysfunction and ventricular remodeling, which would be due to suppressed cardiac hypertrophy, fibrosis, apoptosis and metabolic abnormality.
Subject(s)
Cardiotonic Agents/pharmacology , Endomyocardial Fibrosis/prevention & control , Flavonoids/pharmacology , Hypertrophy, Left Ventricular/prevention & control , Myocytes, Cardiac/drug effects , Ventricular Dysfunction, Left/prevention & control , Animals , Cell Line , Cell Size , Disease Models, Animal , Endomyocardial Fibrosis/etiology , Endomyocardial Fibrosis/mortality , Endomyocardial Fibrosis/pathology , Gene Expression Regulation , Hypertrophy, Left Ventricular/etiology , Hypertrophy, Left Ventricular/mortality , Hypertrophy, Left Ventricular/pathology , Male , Mice , Mice, Inbred C57BL , Myocardium/metabolism , Myocardium/pathology , Myocytes, Cardiac/metabolism , Myocytes, Cardiac/pathology , PPAR alpha/agonists , PPAR alpha/genetics , PPAR alpha/metabolism , PPAR delta/agonists , PPAR delta/genetics , PPAR delta/metabolism , PPAR-beta/agonists , PPAR-beta/genetics , PPAR-beta/metabolism , Pressure/adverse effects , Signal Transduction , Survival Analysis , Ventricular Dysfunction, Left/etiology , Ventricular Dysfunction, Left/mortality , Ventricular Dysfunction, Left/pathologyABSTRACT
BACKGROUND: Our previous study showed that the combined Chinese herbs containing scutellaria baicalensis georgi and gardenia jasminoids ellis inhibited atherosclerosis. In this study, we sought to determine if baicalin and geniposide could inhibit atherosclerosis through Wnt1 and dickkopf-related protein-1 (DKK1). METHODS: The wild-type and ApoE-/- mice were treated with baicalin, geniposide, and baicalin plus geniposide daily by gavage for 12 weeks. Blood lipid levels were measured with an automatic biochemistry analyzer. Aortic atherosclerotic lesion areas were analyzed with Image-ProPlus software. The mRNA and protein expression of DKK1, Wnt1 and nuclear factor-κB (NF-κB) were measured with RT-PCR and Western Blot. Serum levels of interleukin-12 (IL-12) were quantified with ELISA. RESULTS: The baicalin or geniposide monotherapy as well as combination therapy inhibited the development of atherosclerotic lesions, increased Wnt1 and decreased DKK1 expression and elevated the ratio of Wnt1/DKK1 compared with high-lipid diet group. However, only baicalin or geniposide monotherapy decreased NF-κB expression. Moreover, baicalin and geniposide mono- or combination therapy lowered IL-12 levels. Geniposide reduced both serum total cholesterol and low density lipoprotein levels, while baicalin either alone or in combination with geniposide did not affect serum lipid levels. In human, umbilical vein endothelial cells stimulated by oxidized low density lipoprotein, baicalin and geniposide also increased Wnt1 and decreased DKK1 expression and elevated the ratio of Wnt1/DKK1. CONCLUSIONS: Baicalin and geniposide exert inflammation-regulatory effects and may prevent atherosclerotic lesions through enhancing Wnt1 and inhibiting DKK1 expression.
ABSTRACT
Vascular calcification (VC) is widely considered to be a crucial clinical indicator of cardiovascular disease. Recently, certain properties of mesenchymal stem cells (MSCs) have been hypothesized to have potential in treating cardiovascular diseases. However, their effect on the initiation and progression of VC remains controversial. The present study aimed to investigate whether MSCs indirectly mediate VC and their impact on the Wnt signaling pathways. A Transwell system was selected to establish the indirect coculture environment, and hence, vascular smooth muscle cells (VSMCs) were indirectly cocultured in the presence or absence of MSCs at a ratio of 1:1. Osteogenic medium (OS) was added to imitate a calcifying environment. Fourteen days later, VSMCs in the lower layers of the Transwell plates were harvested. Alkaline phosphatase activity and calcium nodules were markedly increased in calcific VSMCs induced by OS. However, these parameters were significantly decreased in VSMCs by indirectly coculturing with MSCs in the same medium. Furthermore, the messenger RNA expression levels of osteopontin and osteoprotegerin were notably increased in VSMCs cultured in OS, but reduced by indirect interaction with MSCs. In addition, the activities of canonical and noncanonical Wnt ligands, winglesstype MMTV integration site family, number 5A (Wnt5a), receptor tyrosine kinaselike orphan receptor 2 (Ror2) and ßcatenin, which are important in the process of VC, were downregulated by indirect contact with MSCs in OS. Thus, indirect coculture with MSCs inhibits VC and downregulates the Wnt signaling pathways.
Subject(s)
Bone Marrow Cells/metabolism , Down-Regulation , Mesenchymal Stem Cells/metabolism , Myocytes, Smooth Muscle/metabolism , Vascular Calcification/metabolism , Wnt Signaling Pathway , Animals , Bone Marrow Cells/pathology , Coculture Techniques , Male , Mesenchymal Stem Cells/pathology , Myocytes, Smooth Muscle/pathology , Rats , Vascular Calcification/pathologyABSTRACT
Osteoclast-like cells are known to inhibit arterial calcification. Receptor activator of NF-κB ligand (RANKL) is likely to act as an inducer of osteoclast-like cell differentiation. However, several studies have shown that RANKL promotes arterial calcification rather than inhibiting arterial calcification. The present study was conducted in order to investigate and elucidate this paradox. Firstly, RANKL was added into the media, and the monocyte precursor cells were cultured. Morphological observation and Tartrate resistant acid phosphatase (TRAP) staining were used to assess whether RANKL could induce the monocyte precursor cells to differentiate into osteoclast-like cells. During arterial calcification, in vivo and in vitro expression of RANKL and its inhibitor, osteoprotegerin (OPG), was detected by real-time PCR. The extent of osteoclast-like cell differentiation was also assessed. It was found RANKL could induce osteoclast-like cell differentiation. There was no in vivo or in vitro expression of osteoclast-like cells in the early stage of calcification. At that time, the ratio of RANKL to OPG was very low. In the late stage of calcification, a small amount of osteoclast-like cell expression coincided with a relatively high ratio of RANKL to OPG. According to the results, the ratio of RANKL to OPG was very low during most of the arterial calcification period. This made it possible for OPG to completely inhibit RANKL-induced osteoclast-like cell differentiation. This likely explains why RANKL had the ability to induce osteoclast-like cell differentiation but acted as a promoter of calcification instead.
Subject(s)
Aorta/metabolism , Myocytes, Smooth Muscle/metabolism , Osteoclasts/metabolism , Osteoprotegerin/genetics , RANK Ligand/genetics , Vascular Calcification/genetics , Acid Phosphatase/genetics , Acid Phosphatase/metabolism , Animals , Aorta/drug effects , Aorta/pathology , Cell Differentiation , Coculture Techniques , Gene Expression Regulation , Isoenzymes/genetics , Isoenzymes/metabolism , Male , Monocytes/cytology , Monocytes/drug effects , Monocytes/metabolism , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/pathology , Osteoclasts/drug effects , Osteoclasts/pathology , Osteoprotegerin/metabolism , RANK Ligand/metabolism , RANK Ligand/pharmacology , Rats , Rats, Sprague-Dawley , Signal Transduction , Tartrate-Resistant Acid Phosphatase , Vascular Calcification/metabolism , Vascular Calcification/pathologyABSTRACT
Hyperlipidemia is considered an independent risk factor for renal dysfunction and induces a significant increase in the expression of inflammatory mediators, which can be used to evaluate the degree of renal injury. Baicalin is widely used in traditional Chinese herbal medicine and has multiple pharmacological effects. The present study investigated whether baicalin can attenuate the expression of vascular cell adhesion molecule 1 (VCAM1) via a reduction in the expression of monocyte chemoattractant protein1 (MCP1) and interleukin6 (IL6) in the kidney of apolipoprotein E (ApoE)knockout (KO) mice fed a high cholesterol diet. These mice were used as a model of atherosclerosis and were treated with baicalin (100 mg/kg/day) daily by gavage for a period of 12 weeks. By contrast, wildtype male C57BL/6J mice were fed a standard diet. Blood samples were obtained from the angular veins of the mice to measure the total cholesterol (TC) and the expression levels of VCAM1, MCP1 and IL6 in the kidney tissues of the mice were analyzed using reverse transcription quantitative polymerase chain reaction and western blot analysis. Following oral administration of baicalin, no significant difference was observed in the TC in the baicalin group compared with the high cholesterol diet control group. The TC was significantly higher in the AopEKO mice compared with the wildtype male C57BL/6J mice. The expression levels of VCAM1, MCP1 and IL6 in the kidney tissues of the baicalin group were lower compared with those in the high cholesterol diet control group. The results suggested that baicalin decreased the expression levels of proinflammatory mediators and prevented kidney dysfunction in the ApoEKO mice fed a high cholesterol diet.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Apolipoproteins E/deficiency , Chemokine CCL2/genetics , Diet, High-Fat , Flavonoids/pharmacology , Gene Expression Regulation/drug effects , Interleukin-6/genetics , Animals , Chemokine CCL2/metabolism , Cholesterol/blood , Interleukin-6/metabolism , Kidney/metabolism , Male , Mice , Mice, Knockout , RNA, Messenger/genetics , RNA, Messenger/metabolism , Vascular Cell Adhesion Molecule-1/genetics , Vascular Cell Adhesion Molecule-1/metabolismABSTRACT
Vascular calcification significantly affects the health of the elderly. Increasing evidence proved that vascular calcification is an actively regulated osteogenic process. The osteochondrocytic differentiation of mesenchymal stem cells (MSCs) is a significant step of osteogenic processes. The Wnt pathways has been identified as contributing to the regulation of osteogenic mineralization during development and disease. However, it remains unknown whether these MSCs in the vascular calcification differentiate into normal vascular smooth muscle cells (VSMCs) in vivo in order to treat damaged vascular tissue or into calcified VSMCs to aggravate calcification correlated to the Wnt pathways. Thus, it is necessary to analyze the mechanisms of MSC differentiation in detail. In the present study a cellcell coculturing in vitro system was used to observe MSCs that directly interact with normal or calcified VSMCs during calcification and to investigate the gene expression of the Wnt pathways during the process. Direct cocultures were established by seeding two different cell types, VSMCs or calcified VSMCs, or a mixture of both at ratios of 5,000:5,000 cells/1.7 cm2 onto either gelatincoated 1.7cm2 chamber slides for immunohistochemical analysis or gelatincoated 75cm2 tissue culture flasks for protein or RNA isolation. Osteoblastic differentiation was evaluated by examining the cell morphology and assessing the activity of alkaline phosphatase in the cell lysates by alkaline phosphatase staining. Additionally, the mRNA expression levels of the genes encoding for proteins involved in the Wnt signaling proteins, Wnt5A, LRP6, Ror2, cJunNterminal kinase and ßcatenin, were assessed in each group. The present study demonstrated that Wnts are expressed in the progress of differentiation of MSCs during calcification. MSCs can differentiate into different cell phenotypes when there is direct cellcell contact with VSMCs or calcified VSMCs, and the Wnt5a/Ror2 signaling pathway may be associated with the determination of differentiation of MSCs in this process.
Subject(s)
Bone Marrow Cells/cytology , Mesenchymal Stem Cells/cytology , Vascular Calcification , Wnt Proteins/metabolism , Animals , Cell Differentiation , Cells, Cultured , Coculture Techniques , Humans , JNK Mitogen-Activated Protein Kinases/genetics , JNK Mitogen-Activated Protein Kinases/metabolism , Low Density Lipoprotein Receptor-Related Protein-6/genetics , Low Density Lipoprotein Receptor-Related Protein-6/metabolism , Male , Mesenchymal Stem Cells/metabolism , Myocytes, Smooth Muscle/cytology , Myocytes, Smooth Muscle/metabolism , RNA, Messenger/metabolism , Rats , Receptor Tyrosine Kinase-like Orphan Receptors/genetics , Receptor Tyrosine Kinase-like Orphan Receptors/metabolism , Wnt Proteins/genetics , Wnt Signaling Pathway , Wnt-5a Protein , beta Catenin/genetics , beta Catenin/metabolismABSTRACT
Baicalin and geniposide, which are respectively isolated from Scutellariae radix and Gardenia jasminoides, have been known to exhibit a number of pharmacological effects, including anti-inflammatory and anti-oxidant. Here, we primarily aimed to observe the protective effects of these two Chinese herbs on inhibiting the development of atherosclerosis in apolipoprotein E knockout mice via lipids regulation and immunoregulation. After the ApoE-/- mice with high-cholesterol diet had received 12-weeks׳ oral administration of either baicalin or geniposide (100 mg/kg), atherosclerotic plaque areas in aorta were measured and exhibited a prominent decrease in the treated mice. We then assayed serum lipids levels, serum Treg-cell-associated cytokines (TGF-ß1 and IL-10) and the frequency of splenic Treg cells. We found that geniposide notably decreased serum TC and LDL-c. Both baicalin and geniposide treated mice showed much more splenic Treg cells and the correlated cytokines (TGF-ß1 and IL-10). Foxp3, as the marker of Treg cell, was detected in atherosclerotic lesions, and we found that Foxp3 expression at both mRNA and protein levels was up-regulated in addition to increased Foxp3 positive Treg cells detected by immunohistochemistry in baicalin or geniposide treated mice. In conclusion, baicalin and geniposide up-regulated the expression of foxp3, promoted the number and function of Treg cells and ameliorated the atherosclerotic lesions progression partly through lipids regulation and immunoregulation.
Subject(s)
Atherosclerosis/drug therapy , Flavonoids/pharmacology , Flavonoids/therapeutic use , Iridoids/pharmacology , Iridoids/therapeutic use , Animals , Aorta/immunology , Aorta/pathology , Apolipoproteins E/genetics , Atherosclerosis/blood , Atherosclerosis/immunology , Atherosclerosis/pathology , Forkhead Transcription Factors/genetics , Forkhead Transcription Factors/immunology , Interleukin-10/blood , Lipids/blood , Male , Mice, Inbred C57BL , Mice, Knockout , Myocardium/pathology , Phytotherapy , Spleen/cytology , T-Lymphocytes, Regulatory/drug effects , T-Lymphocytes, Regulatory/immunology , Transforming Growth Factor beta1/bloodABSTRACT
Atherosclerosis is a systemic inflammatory disease characterized by the accumulation of dendritic cells (DCs) and other types of immune cells in atherosclerotic plaque. In this study, baicalin and geniposide were isolated from Scutellaria baicalensis Georgi and Gardenia jasminoids Ellis, which are the plants used in traditional Chinese medicine to treat a variety of inflammatory diseases. We then investigated whether baicalin and geniposide could induce regression of atherosclerotic lesions in ApoE-/- mice fed a high cholesterol diet and used as a model of atherosclerosis. Following model induction, these mice were treated with baicalin (100mg/kg), geniposide (100mg/kg), and then a mixture containing baicalin (100mg/kg) and geniposide (100mg/kg) administered daily by gavage for a period of 12weeks. The combined administration of baicalin and geniposide significantly reduced atherosclerotic lesions, and modulated the phenotype of dendritic cells in bone marrow and atherosclerotic plaque. Geniposide lowered both plasma lipid levels and DC numbers, while baicalin administered either alone or in combination with geniposide did not decrease plasma lipids. Our results suggest that baicalin and geniposide may have immune-regulatory effects and prevent the formation of atherosclerotic lesions by decreasing the DC numbers, and inhibit DC maturation in bone marrow and infiltration into lesions.
Subject(s)
Atherosclerosis/immunology , Dendritic Cells/drug effects , Flavonoids/pharmacology , Iridoids/pharmacology , Administration, Oral , Animals , Aorta, Abdominal/drug effects , Aorta, Abdominal/immunology , Aorta, Thoracic/drug effects , Aorta, Thoracic/immunology , Apolipoproteins E/genetics , Atherosclerosis/blood , Atherosclerosis/drug therapy , Dendritic Cells/immunology , Diet, High-Fat , Flavonoids/therapeutic use , Iridoids/therapeutic use , Lipids/blood , Male , Mice, Inbred C57BL , Mice, KnockoutABSTRACT
OBJECTIVE: Vascular calcification is a common pathobiological process which occurs among the elder population and in patients with diabetes and chronic kidney disease. Osteoprotegerin, a secreted glycoprotein that regulates bone mass, has recently emerged as an important regulator of the development of vascular calcification. However, the mechanism is not fully understood. The purpose of this study is to explore novel signaling mechanisms of osteoprotegerin in the osteoblastic differentiation in rat aortic vascular smooth muscle cells (VSMCs). METHODS AND RESULTS: VSMCs were isolated from thoracic aorta of Sprague Dawley rats. Osteoblastic differentiation of VSMCs was induced by an osteogenic medium. We confirmed by Von Kossa staining and direct cellular calcium measurement that mineralization was significantly increased in VSMCs cultured in osteogenic medium; consistent with an enhanced alkaline phosphatase activity. This osteoblastic differentiation in VSMCs was significantly reduced by the addition of osteoprotegerin in a dose responsive manner. Moreover, we identified, by real-time qPCR and western blotting, that expression of Notch1 and RBP-Jκ were significantly up-regulated in VSMCs cultured in osteogenic medium at both the mRNA and protein levels, these effects were dose-dependently abolished by the treatment of osteoprotegerin. Furthermore, we identified that Msx2, a downstream target of the Notch1/RBP-Jκ signaling, was markedly down-regulated by the treatment of osteoprotegerin. CONCLUSION: Osteoprotegerin inhibits vascular calcification through the down regulation of the Notch1-RBP-Jκ signaling pathway.
Subject(s)
Homeodomain Proteins/metabolism , Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism , Muscle, Smooth, Vascular/metabolism , Myocytes, Smooth Muscle/metabolism , Osteoprotegerin/pharmacology , Receptor, Notch1/metabolism , Signal Transduction/drug effects , Vascular Calcification/drug therapy , Alkaline Phosphatase/metabolism , Animals , Male , Myocytes, Smooth Muscle/cytology , Osteoblasts/cytology , Osteoblasts/drug effects , RatsABSTRACT
Cardiovascular disease and osteoporosis are major causes of mortality in the elderly. Alendronate, a bisphosphonate, is widely used in the treatment of osteoporosis and may be used to inhibit vascular calcification. However, its mechanisms are not completely understood. The present study aimed to explore novel signaling mechanisms behind the action of alendronate in the osteoblastic differentiation of rat aortic vascular smooth muscle cells (VSMCs). The osteoblastic differentiation of VSMCs was induced by an osteogenic medium. Using von Kossa staining and direct cellular calcium content determination, mineralization was found to be significantly increased in VSMCs induced with osteogenic medium, consistent with an enhanced alkaline phosphatase activity. Osteoblastic differentiation in VSMCs was significantly reduced by the action of alendronate in a dosedependent manner. In addition, the expression of Notch1 and RBPJκ was significantly upregulated in VSMCs cultured with osteogenic medium at the mRNA and protein levels. The effects of Notch1RBPJκ were inhibited by treatment with alendronate in a dosedependent manner. In summary, results of the current study indicate that alendronate inhibits vascular calcification through downregulation of the Notch1RBPJκ signaling pathway.
Subject(s)
Alendronate/pharmacology , Cell Differentiation/physiology , Immunoglobulin J Recombination Signal Sequence-Binding Protein/metabolism , Muscle, Smooth, Vascular/cytology , Myocytes, Smooth Muscle/cytology , Osteogenesis/drug effects , Receptor, Notch1/metabolism , Signal Transduction/drug effects , Alkaline Phosphatase/metabolism , Animals , Calcium/metabolism , Gene Expression Regulation/drug effects , Homeodomain Proteins/genetics , Homeodomain Proteins/metabolism , Male , Myocytes, Smooth Muscle/drug effects , Myocytes, Smooth Muscle/metabolism , Myocytes, Smooth Muscle/pathology , Osteogenesis/genetics , Rats , Vascular Calcification/metabolismABSTRACT
Accumulating evidence have demonstrated that mesenchymal stem cells (MSCs) are involved in the initiation and progression of various vascular diseases. Canonical Wnt signaling controls the fate of MSCs, and plays an important role in vascular calcification. However, vascular calcification can be inhibited by the non-canonical Wnt signaling pathway Wnt5a/Ror2. This study aimed to investigate whether the Wnt5a/Ror2 pathway is associated with determination of the differentiation fate of MSCs in vascular calcification. Direct co-cultures were established by seeding smooth muscle cells (SMCs) or calcified SMCs and MSCs together at ratios of SMCs or calcified SMCs 15x104; SMCs or calcified SMCs 5x104: MSCs 10x104, SMCs or calcified SMCs 10x104: MSCs 5x104. Osteosynthesis-inducing medium (OS) was added to the culture medium in the groups of MSCs with non-calcified SMCs. Cells were cultured for nine days. Osteoblastic differentiation was evaluated by cell morphology and the activity of alkaline phosphatase (ALP) in cell lysates and ALP staining. Furthermore, we investigated the inhibition of Wnt signaling, and observed that the members of the non-canonical signaling pathway Wnt5a/Ror2 were expressed in each group. Additionally, MSCs cultured in culture media with OS did not differentiate into an osteoblast phenotype when in direct contact with non-calcified SMCs, irrespective of the number of MSCs. However, an osteoblast phenotype was observed when MSCs were cultured in media without OS differentiation towards direct contact with calcified SMCs, and the levels of osteoblastic markers had a direct correlation with the number of MSCs. Of note, the Wnt5a protein was associated with the levels of calcification, thus, although rarely detected in non-calcification, Ror2 mRNA in the non-calcified groups was significantly higher compared to that in the calcified groups. Therefore, the Wnt5a/Ror2 pathway is associated with determination of the differentiation fate of bone marrow mesenchymal stem cells in vascular calcification.
Subject(s)
Mesenchymal Stem Cells/metabolism , Myocytes, Smooth Muscle/metabolism , Receptor Tyrosine Kinase-like Orphan Receptors/metabolism , Vascular Calcification/metabolism , Wnt Proteins/metabolism , Alkaline Phosphatase/metabolism , Animals , Bone Marrow Cells/metabolism , Calcification, Physiologic , Cell Communication , Cell Differentiation , Cells, Cultured , Coculture Techniques , Male , Osteoblasts/cytology , Osteoblasts/metabolism , Osteogenesis , RNA, Messenger/biosynthesis , Rats , Receptor Tyrosine Kinase-like Orphan Receptors/genetics , Wnt Signaling Pathway , Wnt-5a ProteinABSTRACT
Recent studies showed that activation of Wnt/ß-catenin pathway promoted the differentiation of osteoblast-like cells in the arterial calcification, but its mechanism remains unknown. In this study, the hypothesis that Wnt/ß-catenin pathway promotes the differentiation of osteoblast-like cells by upregulating the expression of receptor activator of NF-κB ligand (RANKL) was examined. LiCl was used to activate the Wnt/ß-catenin pathway. The differentiation of osteoblast-like cells was observed by Von Kossa staining, calcium content assay, alkaline phosphatase (ALP) activity assay, and detection of osteocalcin expression. Real-time PCR was performed to detect the expression of RANKL and osteoprotegerin (OPG, the decoy receptor of RANKL) during the osteoblast-like cell differentiation. Different concentrations of OPG were added to the culture media respectively to inhibit the function of RANKL, and the change in the differentiation of osteoblast-like cells was evaluated. The results showed that when the Wnt/ß-catenin pathway was activated by LiCl, the expression of RANKL was significantly increased, which coincided with the differentiation of osteoblast-like cells (P<0.05), and the OPG treatment could partly attenuate the promoting effect of Wnt/ß-catenin pathway on the differentiation of osteoblast-like cells (P<0.05), but it failed to completely abolish such effect. It was concluded that activation of Wnt/ß-catenin pathway promotes the differentiation of osteoblast-like cells by both RANKL-dependent and RANKL-independent mechanisms.
Subject(s)
Cell Differentiation/drug effects , Osteoblasts/drug effects , RANK Ligand/metabolism , Signal Transduction/drug effects , Wnt Signaling Pathway/drug effects , beta Catenin/metabolism , Animals , Cells, Cultured , Osteoblasts/metabolism , Osteogenesis/drug effects , RatsABSTRACT
This study investigates the effects of beriberine on the expression of lectin-like ox-LDL receptor-1 (LOX-1), scavenger receptor A (SR-A), SR class B type I (SR-BI) and ATP-binding cassette transporter A1 (ABCA1) in human macrophage-derived foam cells induced by ox-LDL. Different concentrations of Berberine were co-cultured with THP-1 derived foam cells. The mRNA and protein expressions of LOX-1, SR-A, SR-BI and ABCA1 were determined by RT-PCR and Western blot analysis, respectively. Ox-LDL significantly increased the expression of LOX-1 and inhibited the expression of SR-BI in a dose- and time-dependent manner. Berberine significantly inhibited the effects of ox-LDL in a dose- and time-dependent manner. Moreover, ox-LDL significantly promoted ABCA1 expression. However, berberine had no effect on SR-A or ABCA1 expression. Berberine can inhibit the expression of LOX-1 and promote the expression of SR-BI in macrophage-derived foam cells. Therefore, berberine could be used to treat atherosclerotic diseases.
Subject(s)
Atherosclerosis/drug therapy , Berberine/pharmacology , Drugs, Chinese Herbal/pharmacology , Foam Cells/drug effects , Macrophages/drug effects , Receptors, Oxidized LDL/metabolism , Scavenger Receptors, Class B/antagonists & inhibitors , ATP-Binding Cassette Transporters/genetics , ATP-Binding Cassette Transporters/metabolism , Atherosclerosis/metabolism , Atherosclerosis/pathology , Berberine/therapeutic use , Blotting, Western , Cell Line , Coptis/chemistry , Dose-Response Relationship, Drug , Foam Cells/metabolism , Humans , Lipoproteins, LDL/metabolism , Lipoproteins, LDL/pharmacology , Macrophages/metabolism , Phytotherapy , RNA, Messenger/metabolism , Receptors, Oxidized LDL/genetics , Reverse Transcriptase Polymerase Chain Reaction , Rhizome , Scavenger Receptors, Class A/genetics , Scavenger Receptors, Class A/metabolism , Scavenger Receptors, Class B/geneticsABSTRACT
Congenital long QT syndrome is characterized by a prolongation of ventricular repolarization and recurrent episodes of life-threatening ventricular tachyarrhythmias, often leading to sudden death. We previously identified a missense mutation F275S located within the S5 transmembrane domain of the KCNQ1 ion channel in a Chinese family with long QT syndrome. We used oocyte expression of the KCNQ1 polypeptide to study the effects of the F275S mutation on channel properties. Expression of the F275 mutant, or co-expression with the wild-type S275 polypeptide, significantly decreased channel current amplitudes. Moreover, the F275S substitution decreased the rates of channel activation and deactivation. In transfected HEK293 cells fluorescence microscopy revealed that the F275S mutation perturbed the subcelluar localization of the ion channel. These results indicate that the F275S KCNQ1 mutation leads to impaired polypeptide trafficking that in turn leads to reduction of channel ion currents and altered gating kinetics.
Subject(s)
Endoplasmic Reticulum/metabolism , Long QT Syndrome/metabolism , Animals , Cell Line , Humans , KCNQ1 Potassium Channel/genetics , KCNQ1 Potassium Channel/metabolism , Long QT Syndrome/genetics , Mutation, Missense , Phenylalanine/genetics , Phenylalanine/metabolism , Protein Transport/genetics , Serine/genetics , Serine/metabolismABSTRACT
OBJECTIVE: To investigate the effects of the micrometer compound rhizoma coptidis on inflammatory factors and its possible mechanism in rabbit fed with high-lipid food. METHOD: The levels of CRP, IL-1 and TNF-alpha were all determinated by ELISA method. The mRNA and activity of NF-kappaB were determinated by RT-PCR and EMSA, respectively. RESULT: The level of CRP, IL-1 and TNF-alpha were significantly increased by feeding for 16 weeks with high-lipid diet in rabbit. It was significantly increased that the mRNA and the binging activity with DNA of NF-kappaB in thorax aorta of rabbits fed by high-lipid diet, too. The micrometer compound rhizoma coptidis can reverse the effects of high-lipid diet on CRP, IL-1, TNF-alpha and NF-kappaB. CONCLUSION: The micrometer compound rhizoma coptidis can inhibit the expression of inflammatory factor possibly through inhibitting the expression and activity of NF-kappaB.