Abstract:The abnormality of reverse cholesterol transport (RCT) induces the disorder of lipid metabolism, which is a key factor in the pathogenesis of atherosclerosis. It is important to explore the roles and regulatory mechanisms of key proteins mediated RCT in lipid metabolism. These are also important significance to elucidate the molecular mechanism of atherosclerosis. In this topic collection papers, we discuss the effects of liraglutide, momordicin and growth differentiation factor 11 on cholesterol efflux and the expressions of key proteins ATP-binding cassette transporter A1 (ABCA1), ABCG1 and scavenger receptor class B type Ⅰ in RCT. From different angles, we explain the molecular mechanism of RCT key proteins involved in the regulation of cholesterol efflux by drugs or small molecules.

Abstract:Aim To explore the effect of growth differentiation factor 11 (GDF11) on macrophage reverse cholesterol transport and uncover the potential mechanism. Methods Mouse peritoneal macrophages were treated with oxidized low density lipoprotein (ox-LDL), GDF11 and activin receptor-like kinase 7 (ALK7) inhibitor SB431542 for 24 hours. The lipid accumulation was observed with oil red O staining, and the mRNA and protein expression levels of GDF11, ABCA1 and ABCG1 were determined by real-time PCR and Western blot. Mice were given intraperitoneal injection of exogenous GDF11. Peritoneal macrophages were labeled with ³H-cholesterol, then the labeled macrophages were injected into mice intraperitoneally. The mice feces were collected every 8 hours and the liver and blood samples of mice were gathered after 48 hours since cell injection. The radioactivity of ³H was detected to measure the reverse cholesterol transport level. Results After treated with ox-LDL for 24 hours, ox-LDL induced the accumulation of lipid in macrophage and inhibited the GDF11 mRNA and protein expression level. GDF11 treatment effectively inhibited the lipid accumulation in macrophage induced by ox-LDL. Exogenous GDF11 effectively induced macrophage ABCA1 mRNA expression and increased the cholesterol reverse transport level in vivo. However, after treated with GDF11 and ALK7 inhibitor SB431542, the inhibitory effect of GDF11 on intracellular lipid accumulation induced by ox-LDL was antagonized, and the upregulation of ABCA1 expression was also inhibited. Conclusion GDF11 regulates the expression of ABCA1 through ALK7, thus promoting the reverse transport of cholesterol in macrophage.

Abstract:Aim The previous work showed that momordicin(MD28) can up-regulate the expression of ATP-binding cassette transporter A1 (ABCA1) in THP-1-derived foam cells and reduce the intracellular lipid accumulation, but its mechanism is unclear. This study is to analyze the mechanism of action from the post-transcriptional level. Methods MiRNA chip was used to analyze the microRNA (miRNA) that were down-regulated by 1.5-fold in the miRNA of THP-1-derived foam cells treated with 50 mg/L ox-LDL for 48 h after the intervention of MD28 and compared with the genecards miRNA which regulates the ABCA1 gene expression. Of common miRNA, target gene relationship between this miRNA and ABCA1 was verified by the luciferase reporter gene system. qRT-PCR was used to detect ABCA1 mRNA and miR-23b-3p expression levels, protein was detected by Western blot, oil red O staining was used to detect intracellular lipid accumulation. Results miR-23b-3p was the intersection of miRNA chip and genecards. The target gene validation experiment confirmed that ABCA1 was the target gene of miR-23b-3p. MD28 dose-dependently (0 g/L, 0.4 g/L, 1.2 g/L, 3.6 g/L, 5 g/L) and time-dependently (0 h, 6 h, 12 h, 24 h, 48 h) up-regulated ABCA1 expression, the highest expression level of ABCA1 was at 1.2 g/L for 12 h of MD28. ox-LDL up-regulated the expression of miR-23b-3p and was inhibited by MD28, while MD28 decreased intracellular lipid accumulation, and the inhibitor of miR-23b-3p antagonized the effect of MD28 on ABCA1 expression and intracellular lipid accumulation. Conclusion MD28 can up-regulate the expression of ABCA1 and decrease the intracellular lipid accumulation in THP-1 macrophage-derived foam cells by decreasing the expression of miR-23b-3p.

Abstract:Hyperlipidemia is a common risk factor for cardiovascular disease and diabetes. However, the clinical use of statins to control blood lipids increases the risk of new onset diabetes in patients. Conversely, patients with familial hypercholesterolemia (FH) have higher blood lipids and long-term use of statins, which reduce the risk of diabetes. Considering that FH patient is mainly caused by low density lipoprotein receptor (LDLR) mutation, this makes it possible for LDLR mediated cholesterol metabolism to be involved in the onset of diabetes. Statins can upregulate LDLR expression in liver and some tissues and promote cholesterol influx, which may cause some lipotoxicity in some pancreatic islet cells. Pancreatic β cells are more sensitive to cholesterol, and a small amount of cholesterol accumulation can cause cell dysfunction. The purpose of this paper is to discuss the pathological role of LDLR in statins induced diabetes, and to provide new thinking for reducing the adverse reaction of statins and promoting the prevention and treatment of type 2 diabetes.

Abstract:Aim To investigate the effects of liraglutide on cholesterol efflux in HepG2 cells under high glucose condition, and to detect the expression of related protein and further to explore its mechanism of action. Methods HepG2 cells were cultured in vitro, stimulating with high glucose and intervening with different concentrations of liraglutide.Effect of liraglutide on cholesterol efflux in HepG2 cells under high glucose condition was detected by using BODIPY-cholesterol fluorescence assay. Western blot was used to analyze the protein expressions of ATP-binding cassette transporter A1 (ABCA1), ABCG1 and scavenger receptor B1 and the change of extracellular regulated protein kinase 1/2 (ERK1/2) signaling pathway. Results Liraglutide could significantly reduce the fluorescence density of HepG2 cells in high glucose (50 mmol/L) condition, increase the expression of ABCA1 protein and enhance the phosphorylation level of ERK1/2.Conclusion Liraglutide can promote cholesterol efflux in HepG2 cells under high glucose condition, and its mechanism may be related to the regulation of ERK1/2 signaling pathway and the up-regulation of ABCA1 expression.

Abstract:Aim To investigate the effects of exercise training on the function of the CaL channels in cerebral arterial smooth muscle cells of aging spontaneously hypertensive rats. Methods 18 3-month-old male Wistar-Kyoto (WKY) rats and 18 spontaneously hypertensive rats (SHR) were used, besides, 16-month-old male WKY and SHR were randomly assigned into either a sedentary group or an exercise-trained group, each group contained 18 rats. The rats in exercise-trained groups ran on a motor treadmill (60 min/d, 5 d/w) at 16~18 m/min for 8 weeks. After 8 weeks training, body weight and heart weight among these groups were measured. The CaL channel function and the expression of CaL channel α1C protein in cerebral artery were measured by whole-cell patch-clamp technique and Western blot. Results(1)Aging effects:Both heart rate and systolic blood pressure in SHR were much higher than those of the same age WKY rats, the systolic blood pressure in 16-month-old rats were significantly higher than the same strain in 3-month-old rats; The CaL channel current density and CaL channel α1C protein expression in young SHR were extremely increased compared with WKY, whereas aging markedly decreased CaL channel current density and CaL channel α1C protein expression in both hypertensive and normotensive rats with no significant differences between them. (2) Exercise effects:The heart weight index (heart weight/ body weight) in aging exercise-trained groups were markedly higher than those in sedentary strains; Exercise training greatly lowered the systolic blood pressure, and increased CaL channel current density and CaL channel α1C protein expression in SHR. Conclusion Aging reduced CaL channel function of cerebral artery from SHR, while exercise training attenuated remodeling of CaL channel.

Abstract:Aim To investigate the effect of transfection of recombinant adenovirus vector carrying angiotensin converting enzyme 2 (ACE2) gene (Ad-ACE2) on protecting endothelium, improving insulin resistance and inhibiting liver fibrosis in apolipoprotein E knock-out (ApoE-/-) mice fed with high-fat diet. Methods ApoE-/- mice were fed with high-fat and high-energy food to simulate metabolic syndrome. 30 ApoE-/- mice were randomly divided into control group (Con group), adenovirus empty vector group (EGFP group) and ACE2 gene therapy group (ACE2 group), with 10 mice in each group, and each group was given a high-fat diet. The 3 groups were injected respectively with normal saline, Ad-EGFP and Ad-ACE2 by intravenous injection of the tail vein. At the end of 12 weeks, glucose tolerance test and insulin tolerance test were carried out. After the mice anaesthesia, blood was extracted from heart tip and blood lipid level was measured. Liver tissue was taken with HE staining and oil red O staining. Immunohistochemical staining was used to observe the expressions of ACE2, collagen-Ⅰ (COL-Ⅰ), collagen-Ⅲ (COL-Ⅲ) and interleukin-6 (IL-6) in liver tissue. Results In ACE2 group, Ad-ACE2 transfection significantly increased the expression of ACE2 in ApoE-/- mice liver (P＜0.05). The endothelium-dependent function in ACE2 group was significantly better than that in Con group and EGFP group (P＜0.05), and insulin sensitivity was stronger than that in Con group and EGFP group (P＜0.05), but the level of blood lipid did not change significantly. The COL-Ⅰ, COL-Ⅲ and IL-6 expressions of liver tissue in ACE2 group were lower than those in Con group and EGFP group (P＜0.05). Conclusion ACE2 overexpression can protect the endothelium, improve insulin resistance, inhibit liver fibrosis and thus protect liver function.

Abstract:Aim To investigate the effect of high salt on cardiac fibroblast proliferation and intracellular free Ca²＋ concentration and pH. Methods SD rat cardiac fibroblasts were cultured by adherent method, and vimentin in cardiac fibroblasts was identified by immunofluorescence staining. The third generation cells were divided into control group (Na＋ 122.27 mmol/L) and high salt group (Na＋ 161 mmol/L), treated for 48 hours. Cell proliferation was detected by EdU fluorescent staining, the treated cells were loaded with Fura2-AM and BCECF-AM fluorescent indicators to measure intracellular pH and free Ca²＋concentration. Results The proliferation of cardiac fibroblasts in the high salt group was significantly higher than that in the control group, and the intracellular pH and free Ca²＋ concentration in the high salt group were significantly higher than those in the control group as well. Conclusion High salt cloud induce cardiac fibroblast proliferation and lead to increase intracellular free Ca²＋ concentration and pH value.

Abstract:Aim To investigate the antagonistic effect of endostatin (ENDO)-vascular endothelial growth inhibitor (VEGI) recombinant adenoviruses to the vascular endothelial cell injury caused by homocysteine (Hcy). Methods The Ad-hENDO-VEGI151 from the fusion protein of ENDO and VEGI was used to transfect the human ECV304 caused by Hcy. The expression of fusion protein was detected by Western blot, the leakage of lactate dehydrogenase (LDH) was detected by automatic biochemical analyzer, the cell viability was detected by trypan-blue staining method, and the expression of tumor necrosis factor-α (TNF-α) and monocyte chemoattractant protein-1 (MCP-1) were detected by ELISA. Results The transgenic efficiency of recombinant adenovirus was relatively high. A fusion protein with molecular weight of 41 kDa was expressed in the Ad-hENDO-VEGI151 recombinant adenovirus of ECV304 cells. Hcy (0.1~1.0 mmol/L) increased the leakage of LDH and decreased the cell viability in concentration-dependent manner. The recombinant adenovirus effectively inhibited the injury of Hcy on human vascular endothelial cells (P＜0.05). Conclusion The ENDO-VEGI recombinant adenoviruses had antagonistic effect on vascular endothelial cell injury caused by Hcy, which might provide an idea for future prospecting.

Abstract:Aim To study the preventive effect and mechanism of ligustrazine on doxorubicin-induced cardiotoxicity in mice. Methods The mouse model of cardiotoxicity induced by doxorubicin was established. The mice were randomly divided into three groups:sham group, doxorubicin group (doxorubicin 15 mg/kg), ligustrazine treated group (ligustrazine 60 mg/(kg·d)＋doxorubicin 15 mg/kg), echocardiography was performed to detect cardiac function of doxorubicin and ligustrazine intervention groups in mice. The sections were stained with HE staining or Masson's trichrome staining for histological and collagen analysis, apoptosis of cardiomyocyte were analyzed by TUNEL staining, Western blot was used to analyze the expression of Akt, endothelial nitric oxide synthase (eNOS), the phosphorylation of Akt and eNOS.Results Compared with doxorubicin group, the fractional shortening (FS) and the ventricular ejection fraction (EF) in the ligustrazine treated group were significantly higher (P＜0.01). The ligustrazine treated group significantly decreased apoptosis of cardiomyocyte in mice as compared to the doxorubicin group (P＜0.01), the expression of p-Akt and p-eNOS in the ligustrazine treated group were stronger than those in the the doxorubicin group (P＜0.01). Meanwhile, the ligustrazine treated group significantly inhibited cleaved Caspase-3 as compared to the doxorubicin group. Conclusion Ligustrazine protects doxorubicin-induced myocardial injury by activating Akt/eNOS signaling pathway and inhibiting cardiomyocyte apoptosis, and it may be an underlying mechanism by which ligustrazine can prevent doxorubicin-induced cardiotoxicity.

Abstract:Aim To explore the circulating levels of long intergenic non-coding RNA predicting cardiac remodeling (LIPCAR) in patients with chronic heart failure (CHF) and combined renal insufficiency. Methods Venous blood samples were collected from 80 control subjects, 188 patients with CHF, including 80 patients with renal insufficiency, LIPCAR was evaluated by real-time PCR. Results The expression of LIPCAR was significantly different between the control group and the CHF group (1.08±0.28 vs.0.79±0.31, P＜0.01). In addition, the subgroup analysis of CHF showed that the patients complicated with renal insufficiency exhibited a tendency of lower levels of LIPCAR. Furthermore, LIPCAR was negatively correlated with NT-proBNP, whereas positively correlated with ejection fraction (P＜0.01). AUC was 0.777(95%CI＝0.720～0.835) for LIPCAR to judge CHF. Conclusion Circulating LIPCAR levels were significantly downregulated in patients with CHF, especially in those complicated with renal insufficiency. It may be valuable as a potential biomarker for CHF.

Abstract:Aim To find risk factors of contrast-induced nephropathy (CIN) after percutaneous coronary intervention (PCI) in elderly diabetic patients with acute coronary syndrome (ACS). Methods 134 cases of elderly diabetic patients with ACS (≥65 years old) was involved in this study. In the same period, 145 cases of PCI patients without diabetes were used as controls. The age, preoperation brain natriuretic peptide (BNP), ejection fraction, 25 hydroxyvitamin D, estimated glomerular filtration rate (eGFR), blood albumin and so on were compared between the two groups. Multifactor logistic regression analysis was used to find risk factors of CIN. Results 17 cases of CIN were found in 134 cases of patients with diabetes after PCI, the incidence was 12.6%. In 145 elderly patients without diabetes, there were 13 cases of CIN, with an incidence of 8.9%, there was no statistically significant difference between the two groups (P＞0.05). Statistically difference was found in gender and perioperative use of ACEI/ARB between two groups (P<0.05). Preoperation BNP, fibrinogen, platelet volume distribution of the diabetes group was higher than non-diabetic, 25 hydroxyvitamin D was lower than non-diabetic group (P<0.05). Loistic regression analysis showed that fibrinogen and 25 hydroxyvitamin D were the risk factors of CIN for elderly diabetic patients with acute coronary syndrome after PCI. Preoperative fibrinogen elevation increased the risk of CIN (OR=3.3,5%CI 1.353～6.845, P=0.007), 25 hydroxyvitamin D reduction increased the risk of CIN (OR=0.5,5%CI 0.282～0.833, P=0.009). Conclusion The risk of CIN was increased when preoperative fibrin was elevated and 25 hydroxyvitamin D was decreased in elderly diabetes mellitus patients with acute coronary syndrome after PCI.

Abstract:Aim To investigate therapeutic value and effectiveness evaluation of the continuous renal replacement therapy (CRRT) hemoperfusion combined with continuous veno-venous hemofiltration(HP/CVVH), plasma exchange combined with continuous veno-venous hemofiltration (PE/CVVH) on hyperlipidemic patients with severe acute pancreatitis (HLSAP). Methods 68 patients were randomly divided into CVVH group, HP+CVVH group, PE+CVVH group, herein are SAP and conventional therapy alone CVVH, HP+CVVH, PE+CVVH treatment. The vital signs, APACHE II score, triglyceride (TG),Cü reactive protein (CRP) and important organ function indexes of the three groups were compared before and after treatment for 72 h; The CT severity index (CTSI) change was evaluated by CT examination before and after 1 week of treatment; The incidence of major complications, average length of stay and mortality were recorded within 1 week of treatment. Results There was no significant difference between the three groups before treatment. But all the indexes were improved after treatment in each group(P<0.05). There was no significant difference in APACHE Ⅱ score, serum amylase, and urine amylase between the three groups after treatment for 72 h (P>0.05); Remaining laboratory parameters improved more significantly in HP+CVVH, PE+CVVH groups compared with CVVH group (P<0.05). After 1 week of treatment, the CTSI of three groups decreased compared with before treatment (P<0.05), and the improvement of HP+CVVH group and PE+CVVH group was more obvious than that of CVVH group (P<0.05). Compared with CVVH group, the clinical situation of HP+CVVH group and PE+CVVH group was better, the mortality rate was lower and the length of stay was shorter (P<0.05). Conclusion Compared with CVVH alone, HP, PE combined with CVVH treatment can reduce the serum TG level of patients more quickly, improve clinical symptoms, reduce systemic inflammatory response syndrome (MODS) and multiple organ dysfunction syndrome/multiple organ failure(MOF) occurs, effectively reduce mortality and hospital stay.

Abstract:Aim In order to evaluate the instability of atherosclerotic plaque more comprehensively and objectively, the computational method of plaque vulnerability index based on lipid accumulation, which was put forward by Shiomi was improved in this study. Methods The experimental was divided into the control group (C57BL/6 mice) and the model group (LDLR－/－ mice and ApoE－/－ mice), with 6 mice in each group. The control group was given normal diet, while the model group was given high-fat diet. After 25 weeks, the mice were sacrificed and the aortic root and aortic arch were isolated and sectioned after OCT embedding. Serial sections were stained by hematoxylin and eosin (H&E), picrosirius red, oil red O, alcian blue, and quantitatively studied the amount of endothelial cell and smooth muscle cell by immunofluorescence, as well as matrix metalloproteinase and macrophage with immunohistochemistry. The calculation formula of plaque vulnerability index proposed by Shiomi was perfected through adding the factors affecting the plaque instability to the formula numerator and the stability index of the plaque to the denominator. The stability of aorta root and innominate artery plaque in LDLR－/－ mice was compared by two formulas, and the reliability of the optimized formula was evaluated. Finally, the stability of atherosclerotic plaques in different parts of aorta of ApoE－/－ and LDLR－/－ mice was evaluated by the optimized formula, and the difference of atherosclerosis progression between two genetically engineered mice was compared. Results The plaque damage area, lipid deposition, the distribution of collagen, proteoglycan and matrix metalloproteinase, macrophage infiltration, the endothelial cell coverage and smooth muscle cell coverage of fibrous cap in the model group were significantly changed compared with the control group. On this basis, the formula for calculating plaque instability based on multi index pathological staining was optimized. Through calculation, it was found that Plaque vulnerability index of innominate artery was significantly larger than that of aortic root in ApoE－/－ and LDLR－/－ mice. In the same site, plaque vulnerability index in ApoE－/－ mice was significantly higher than that in LDLR－/－ mice. Conclusion Optimized calculation formula of plaque vulnerability index based on multi index pathological staining covers a number of important factors that affect plaque instability, and can accurately assess plaque instability. The method is simple, comprehensive, reliable and of great practical value.

Abstract:Aim To explore the molecular mechanism of Radix Salviae on atherosclerosis based on the network pharmacology. Methods Radix Salviae’s active compounds were collected from reference and traditional Chinese Medicine databases, and put into PharmMapper to get their potential targets. Then KEGG-pathway analysis and GO enrichment analysis were made by DAVID database. Finally the network was drawn and analyzed by Cytoscape with information above. Results Sixteen compounds of THP acquired 344 compound targets which also got 6 atherosclerosis-related pathways and 28 biological processes. The biological processes can be divided into four modules—protection of endothelial cell function, antioxidant, anti-atherosclerotic plaque formation, conditioning blood lipid levels. Conclusion These findings reflect the molecular mechanism of Radix Salviae on atherosclerosis. It provides not only a theoretical basis for the study of Radix Salviae on atherosclerosis, but also a new paradigm for the study of the mechanism of traditional Chinese medicine such as Radix Salviae.

Abstract:As a newly discovered gas signal molecule after nitric oxide and carbon monoxide, hydrogen sulfide (H₂S) has attracted wide attention from experts and scholars both at home and abroad, and has been carried out a lot of physiological and pathophysiological studies. Most scholars believe that H₂S plays an important role in the development of diabetes and its microvascular disease. However, so far, the physiological and toxic effects of H₂S in the human body are not completely clear, and most of the mechanisms of the H₂S effect have not been fully elucidated. This paper reviews the role and mechanism of H₂S in 3 common diabetic microangiopathy (diabetic retinopathy, diabetic neuropathy and diabetic nephropathy), in order to provide a new theoretical basis for the research and treatment of diabetic microangiopathy.

Abstract:Atherosclerosis is one of the most serious cardiovascular and cerebrovascular diseases threatening human health. It has become a hot topic of medical research to elucidate the mechanism of atherosclerosis and prevent atherosclerosis. Carotid atherosclerosis can reflect the whole body atherosclerosis plaque development in a certain extent, the location of the carotid artery is shallow, and it is easy to detect and examine, which has been proved to be a "window" to understand and evaluate the whole body atherosclerosis. Confirmed by research, endothelial progenitor cells can repair the damaged endometrium, delay the development of atherosclerosis. In this paper, the number and function of endothelial progenitor cells in patients with carotid atherosclerosis and the role of endothelial progenitor cells in the development of the disease are generalized and summarized, providing new strategies for the treatment and prevention of atherosclerosis.

Abstract:Intermedin (IMD) is a new type of cardiovascular regulatory peptide found in recent years. It is a new member of calciton gene related peptide (CGRP). Previous studies have found that the IMD has important protective effect in ischemia reperfusion. Epidemiological and clinical studies have shown that diabetic population of ischemic heart disease (angina and myocardial infarction), congestive heart failure and the incidence of atherosclerotic lesions caused by disease and its severity is much higher than people without diabetes. Therefore, it remains to be discussed whether the IMD has protective effect in diabetic patients with coronary heart disease. This article reviews the biological characteristics of the IMD and its role in diabetic patients combined with coronary heart disease from basic research to clinical research progress.

Abstract:Atherosclerosis is the pathological basis of cardiovascular and cerebrovascular diseases. Chronic inflammatory reaction and endothelial dysfunction play an important role in the occurrence and development of atherosclerosis. In recent years, some new markers of endothelial injury and inflammation have been identified, including Endoglin (CD105), Endocan, Apelin, and chemokines. This article mainly introduces the role of Endoglin in atherosclerosis.