Response of genes involved in lipid metabolism in rat epididymal white adipose tissue to different fasting conditions after long-term fructose consumption.

There has been much concern regarding the dietary fructose contributes to the development of metabolic syndrome. High-fructose diet changes the expression of genes involved in lipid metabolism. Levels of a number of hepatic lipogenic enzymes are increased by a high-carbohydrate diet in fasted-refed model rats/mice. Both the white adipose tissue (WAT) and the liver play a key role in the maintenance of nutrient homeostasis. Here, the aim of this study was to analyze the expression of key genes related to lipid metabolism in epididymal WAT (eWAT) in response to different fasting condition after long-term chronic fructose consumption. Rats were fed standard chow supplemented with 10% w/v fructose solution for 5 weeks, and killed after chow-fasting and fructose withdrawal (fasting) or chow-fasting and continued fructose (fructose alone) for 14 h. Blood parameters and the expression of genes involved in fatty acid synthesis (ChREBP, SREBP-1c, FAS, SCD1), triglyceride biosynthesis (DGAT-1, DGAT-2) and lipid mobilization (ATGL, HSL) in eWAT were analyzed. In addition, mRNA levels of PPAR-γ, CD36 and LPL were also detected. As expected, fructose alone increased the mRNA expression of FAS, SCD1, and correspondingly decreased ATGL and HSL mRNA levels. However, ChREBP, DGAT-2, ATGL and HSL mRNA levels restored near to normal while FAS and SCD1 tend to basic level under fasting condition. The mRNA expression of SREBP-1c, PPAR-γ and LPL did not changed at any situations but CD36 mRNA decreased remarkably in fructose alone group. In conclusion, these findings demonstrate that genes involved in lipid metabolism in rat eWAT are varied in response to different fasting conditions after long-term fructose consumption.

Long-term fructose consumption prolongs hepatic stearoyl-CoA desaturase 1 activity independent of upstream regulation in rats.

Dietary fructose is considered a risk factor for metabolic disorders, such as fatty liver disease. However, the mechanism underlying the effects of fructose is not well characterized. We investigated the hepatic expression of key regulatory genes related to lipid metabolism following fructose feeding under well-defined conditions. Rats were fed standard chow supplemented with 10% w/v fructose solution for 5 weeks, and killed after chow-fasting and fructose withdrawal (fasting) or chow-fasting and continued fructose (fructose alone) for 14 h. Hepatic deposition of triglycerides was found in rats from both groups. As expected, fructose alone increased mRNA levels of lipogenesis-related genes and correspondingly decreased mRNA levels of lipid oxidative genes in the liver. Interesting, hepatic levels of stearoyl-CoA desaturase (SCD)1 mRNA remained elevated under fructose withdrawn conditions, although expression levels of other genes, including two key transcription factors (carbohydrate response element binding protein (ChREBP) and sterol regulatory element-binding protein (SREBP)-1c) fell to normal levels, indicating that long-term fructose intake increased SCD1 activity, independent of upstream regulatory genes, such as ChREBP and SREBP-1c. In conclusion, SCD1 overexpression in fatty liver disease is not affected by fasting after long-term fructose consumption in rats. Regulation of SCD1 plays an important role in fructose-induced hepatic steatosis.

[Ghrelin stimulates in vitro angiogenic capacity of rat cardiac microvascular endothelial cells].

OBJECTIVE: To clarify whether ghrelin could promote in vitro rat cardiac microvascular endothelial cells (CMECs) angiogenesis and related mechanisms. METHODS: CMECs were isolated from myocardial tissue of adult male SD rats and characterized by the immunocytochemistry staining with Factor VIII and the capacity of in vitro capillary tube-like formation. The mRNAs and protein expressions of ghrelin and its receptor (growth hormone secretagogue receptor, GHS-R) of CMECs were determined by RT-PCR, Immunofluorescence, ELISA and Western blot. Proliferation, migration and in vitro angiogenesis as well as ERK2 phosphorylation of CMECs were tested in the presence of ghrelin (10(-9) - 10(-7) mol/L) with or without pretreatment with specific MAPK/ERK2 inhibitor PD98059. RESULTS: Purity of CMECs characterized by immunocytochemistry staining with Factor VIII was about 95%, and the cells showed a high ability to form the capillary tube-like structures on Matrigel. Ghrelin and GHS-R were constitutively expressed in CMECs. Proliferation, migration and in vitro angiogenesis capacities of CMECs (72.20 ± 5.72 vs. 28.60 ± 5.13, P < 0.001; 71.00 ± 7.78 vs. 28.60 ± 5.13, P < 0.001) as well as ERK2 phosphorylation (0.92 ± 0.13 vs. 0.29 ± 0.04, P < 0.001; 1.15 ± 0.16 vs. 0.29 ± 0.04, P < 0.001) were significantly enhanced by exogenous ghrelin (10(-8) - 10(-7) mol/L). PD98059 abolished ghrelin-induced ERK2 phosphorylation and in vitro angiogenesis. CONCLUSIONS: Ghrelin and its receptor are expressed in CMECs and ghrelin could stimulate CMECs in vitro angiogenesis through activation of MAPK/ERK2 signaling pathway.

Suppression of NHE1 by small interfering RNA inhibits HIF-1α-induced angiogenesis in vitro via modulation of calpain activity.

Hypoxia-inducible factor-1 (HIF-1) orchestrates angiogenesis under hypoxic conditions mainly due to increased expression of such target genes as vascular endothelial growth factor (VEGF). Na+/H+exchanger-1 (NHE1), a potential HIF target gene product, plays a pivotal role in proliferation, survival, migration, adhesion and so on. However, it is unknown whether NHE1 is involved in HIF-1α-induced angiogenesis. This present study demonstrated that the expression of NHE1 was much higher in human umbilical vein endothelial cells (HUVECs) infected with adenovirus encoding HIF-1α (rAd-HIF) than with vacuum adenovirus (vAd). HIF-1α also increased the expression of VEGF, the expression and activity of calpains, and the intracellular pH. Moreover, small interfering RNA targeting NHE1 (NHE1 siRNA) dramatically decreased the expression of NHE1 and thus lowered the intracellular pH, and it also attenuated the protein expression of calpain-2 but not calpain-1, resulting in the lower calpain activity. Furthermore, HIF-1α enhanced the proliferation, migration and Matrigel tube formation, which were inhibited by NHE1 siRNA. Finally, the inhibitory effect of NHE1 siRNA was reversed by VEGF and the reversibility of the later was abrogated by the calpain inhibitor ALLM. In conclusion, the findings have revealed that NHE1 might participate in HIF-1-induced angiogenesis due, at least in part, to the alteration of the calpain activity, suggesting that NHE1 as well as calpains might represent a potential target of controlling angiogenesis in response to the hypoxic stress under various pathological conditions.

Bone marrow mesenchymal stromal cells with support of bispecific antibody and ultrasound-mediated microbubbles prevent myocardial fibrosis via the signal transducer and activators of transcription signaling pathway.

BACKGROUND AIMS: This study was initiated to investigate the efficacy of myocardial fibrosis intervention via signal transducer and activators of transcription (STAT) signaling using bone marrow (BM) mesenchymal stromal cells (MSC) with the aid of bispecific antibody (BiAb) and ultrasound-mediated microbubbles (MB). METHODS: BiAb (anti-CD29 × anti-myosin light chain antibody; AMLCA) was prepared and combined with isolated MSC from male mice and transfused into female mice with isoproterenol-induced myocardial fibrosis via the tail vein, followed by MB (MSC + BiAb + MB). This study included seven groups: MSC + BiAb + MB; MSC; BiAb; MB; MSC + BiAb; untreated; and control. Five weeks after treatment, expression levels of the sex-determining region of Y-chromosome (SRY), matrix metalloproteinases (MMP)-9, tissue inhibitor of metalloproteinase (TIMP)-1 and vascular endothelial growth factor (VEGF) in myocardium were detected by fluorescent quantitative real-time polymerase chain reaction (qRT-PCR). Collagen distribution was observed using Sirius Red staining. The protein expression of signal transducer and activators of transcription (STAT)1 and STAT3 was detected by Western blot. RESULTS: The highest homing number of MSC was in the MSC + BiAb + MB group, second highest in the MSC + BiAb group, and lowest in MSC alone. Compared with the untreated group, MSC + BiAb + MB, MSC + BiAb and MSC groups had decreased levels of MMP-9, TIMP-1, STAT1 and collagen deposition, and increased levels of STAT3. Upregulated STAT3 and downregulated TIMP-1 were significantly different in MSC + BiAb + MB compared with MSC alone or MSC + BiAb. CONCLUSIONS: The homing rate and repairing efficacy of MSC improved with treatment utilizing a combination of BiAb and MB. MSC can improve MMP-TIMP expression in injured myocardium and interfere with myocardial fibrosis after homing, a mechanism that may be related to the STAT-mediated signaling pathway.

[Cytokines mechanism of shenfu injection in treatment of cardiogenic shock in canine].

OBJECTIVE: To investigate the effect of shenfu injection on canine with cardiogenic shock and the possible mechanism. METHOD: Cardiogenic shock model of canine was established by ligating left anterior descending (LAD) of coronary artery. The 15 canines with cardiogenic shock were randomly divided in to glucose injection group, shenfu injection group and sham-operated group. The hemodynamics parameters were monitored. Plasma TNF-alpha and IL-1beta levels were measured by radioimmunoassay. Expression of TNF-alpha mRNA and IL-1beta mRNA in myocardium were detected by RT-PCR. RESULT: Following cardiogenic shock, the mean artery pressure (MAP), left ventricular systolic pressure (LVSP), ventricular pressure rise ratio during systolic period (+ dp/dt(max)), and ventricular pressure decay ratio during diastolic period (- dp/dt(max)) decreased significantly; the plasma TNF-alpha and IL-1beta levels and the expression of TNF-a mRNA and IL-1beta mRNA in myocardium increased significantly. In shenfu injection group, MAP, LVSP and +/- dp/dt(max) increased significantly and plasma TNF-alpha and IL-1beta levels decreased significantly. In glucose injection group, MAP, LVSP, +/- dp/dt(max) and plasma TNF-alpha and IL-1beta levels had not changed significantly. The expression of TNF-alpha mRNA and IL-1beta mRNA in myocardium were significantly lower in shenfu injection group than those in glucose injection group. CONCLUSION: Shenfu injection probably can decrease over-exprssion of TNF-alpha mRNA and IL-1beta mRNA on transcription platform. Shenfu injection counteract cardiogenic shock, relieve myocardium damage and improve hemodynamics through inhibiting overproduction of TNF-alpha and IL-1beta.

[Predictive value of fibrinogen and high-sensitivity C-reaction protein for cardiovascular events in patients with stable coronary artery disease].

OBJECTIVE: To investigate the predictive value of elevated fibrinogen and high-sensitivity C-reaction protein (hs-CRP) level on cardiovascular events in patients with stable coronary artery disease (CAD). METHODS: From January 2002 to November 2002, 185 patients (aged 47 - 85 years) with stable CAD referred for coronary angiography were enrolled and divided into control-F (fibrinogen level < or = 4.0 g/L, n = 104) and elevated-F (fibrinogen level > 4.0 g/L, n = 81), or control-hs (hs-CRP < or = 3.0 mg/L, n = 99) and elevated-hs (hs-CRP> 3.0 mg/L, n = 86). Exclusion criteria included cardiomyopathy, New York Heart Association class IV congestive heart failure, recent myocardial infarction or coronary artery revascularization and cancer. During three years follow-up, cardiovascular death, myocardial infarction, congestive heart failure, stroke and other vascular events were assessed. RESULTS: A total of 21 cardiovascular nonfatal events and 10 cardiovascular deaths were observed. Cardiovascular events was significantly higher in patients in elevated-F group than that in control-F group [23.46% vs. 11.54%, cholesterol-, body mass index-, smoking-, and hypertension-adjusted relative risk 1.97, 95% CI (1.68 to 2.40), P < 0.05] and in elevated-hs group than in control-hs group [24.42% vs. 10.10%, adjusted relative risk 2.32, 95% CI (1.76 to 2.89), P < 0.05]. The relative risk of cardiovascular events for patients with fibrinogen > 4.0 g/L and hs-CRP > 3.0 mg/L was 3.84 (P < 0.05), 95% CI (2.80 to 4.99) compared with patients with fibrinogen < or = 4.0 g/L and hs-CRP < or = 3.0 mg/L. CONCLUSION: Both fibrinogen and hs-CRP are independent important predictors of cardiovascular nonfatal and fatal events in patients with stable CAD. Combination of elevated fibrinogen and hs-CRP increased their predictive value for cardiac events.

Bone marrow mesenchymal stromal cells with CD47 high expression via the signal transducer and activators of transcription signaling pathway preventing myocardial fibrosis.

UNLABELLED: This study was initiated to investigate the efficacy of myocardial fibrosis intervention via signal transducer and activators of transcription (STAT) signaling using bone marrow (BM) mesenchymal stromal cells (MSC) in which being over-expressed with the aid of bispecific antibody (BiAb) and ultrasound-mediated microbubbles (MB). BiAb was prepared and combined with isolated MSC with CD47 overexpression from male mice and trans-fused into female mice with isoproterenol-induced myocardial fibrosis via the tail vein, followed by MB. This study included five groups. Five weeks after treatment, expression levels of the sex-determining region of Y-chromosome (SRY), matrix metalloproteinases (MMP)-9, tissue inhibitor of metalloproteinase (TIMP)-1 and vascular endothelial growth factor (VEGF) in myocardium were detected by fluorescent quantitative real-time polymerase chain reaction (qRT-PCR). The protein expression of signal transducer and activators of transcription (STAT) 1 and STAT 3 was detected by Western blot. RESULTS: The highest homing number of MSC was in the CD47 + MSC + BiAb + MB group, second highest in the CD47 + MSC + BiAb group, and lowest in MSC alone. Compared with the Control group, CD47 + MSC + BiAb + MB, CD47 + MSC + BiAb, CD47 + MSC and MSC groups had decreased levels of MMP-9, TIMP-1, STAT 1 and collagen deposition, and increased levels of STAT 3. Up regulated STAT 3 and down regulated TIMP-1 were significantly different in CD47 + MSC + BiAb + MB compared with CD47 + MSC or CD47 + MSC + BiAb. CONCLUSION: CD47 can enhance the homing rate and repairing efficacy of MSC. MSC can improve MMP-TIMP expression in injured myocardium and interfere with myocardial fibrosis after homing, a mechanism that may be related to the STAT-mediated signaling pathway.

Association of circulating endothelial progenitor cells (CD14+-EPC) with renal function in patients with coronary artery disease.

BACKGROUND: The relationships between the endothelial progenitor cells (EPCs)-CD34(+) and CD14(+) and coronary artery disease (CAD) were reported and the association of CD34(+) cells with renal function was studied previously. Another kind EPC-CD14(+) cell and its association with renal function in patients with CAD have not been reported yet. Our aim was to assess CD14(+) cell counts versus renal function in CAD. METHODS AND RESULTS: We studied 242 patients with severe angiographic CAD and 30 healthy control participants. The CD14(+) cells were enumerated by flow cytometry. With lowering glomerular filtration rate (GFR), CD14(+) cell numbers (percentage of lymphocytes, median and interquartile range) decreased: 0.04 (0.03-0.06), 0.03 (0.02-0.05), 0.02 (0.01-0.03) for estimated glomerular filtration rate (eGFR) ≥90, 60 to 89, and 30 to 89 mL/min per 1.73 m(2), respectively (P < .001 for trend). The CD14(+) cell counts correlated with eGFR (r = .27, P = .03). By multivariate liner regression analysis, the difference remains significant (P = .02). CONCLUSIONS: The CD14(+) cell depletion is associated with renal dysfunction in CAD.

Procollagen III N-terminal peptide predicts short-term prognosis and cardiac remodeling in coronary heart disease patients with metabolic syndrome.

INTRODUCTION: Many patients with coronary heart disease (CHD) also have metabolic syndrome (MS); however, little is known about the condition of cardiovascular remodeling in these patients. The objective of this study to explore the role of plasma procollagen III N-terminal peptide (PIIINP) in predicting the prognosis and cardiac remodeling in patients with CHD with MS. METHODS: One hundred eight patients were classified into high and low PIIINP groups according to the median value of plasma PIIINP. Cardiovascular examinations including echocardiogram, carotid color ultrasound examination, coronary angiography and the 6-minute walking test (6MWT) were performed before and after a 1-year follow-up. Readmission for cardiac and cerebrovascular events was assessed during the follow-up period. RESULTS: Plasma PIIINP level was significantly correlated with age, high-sensitivity C-reactive protein (hs-CRP) and body mass index in a multiple stepwise regression model. There was a positive correlation between the LnPIIINP and an increased left ventricular mass index in partial correlation analysis. The Cox proportional hazard model analysis indicated that the level of PIIINP, left ventricular ejection fraction and hs-CRP were independent predictors of readmission owing to cardiac and cerebrovascular events during the follow-up. A PIIINP value of 4.0 µg/L was the best threshold value for determining the need for readmission. CONCLUSIONS: PIIINP levels rise with increases in age, hs-CRP and body mass index in patients with CHD with MS, and a high level of PIIINP indicates recent deterioration of cardiac remodeling and exercise tolerance and a poor prognosis.