Obestatin Plays Beneficial Role in Cardiomyocyte Injury Induced by Ischemia-Reperfusion In Vivo and In Vitro.

BACKGROUND Obestatin, primarily recognized as a peptide within the gastrointestinal system, has been shown to benefit the cardiovascular system. We designed this experiment to study the protective role and underlying mechanism of obestatin against ischemia-reperfusion(I/R) injury in myocardial cells. MATERIAL AND METHODS In an In vivo experiment, LAD was ligated for 0.5 h and then opened for reperfusion with obestatin for 24 h. Then, the infarction area was shown with TTC staining, and inflammation factors in serum were analyzed by qRT-PCR. In primary cultured cardiomyocytes, we measured the level of LDH, MDA, GSH, and SOD. Finally, we assessed cells apoptosis using flow cytometry and detected the concentrations of caspase-3, Bax, and Bcl-2 using Western blot analysis. RESULTS TTC staining showed that in the 3 obestatin groups, the infarct area became smaller with the increase of obestatin concentration. Obestatin also inhibited LDH expression in rat serum and decreased mRNA levels of TNF-α, IL-6, ICAM-1, and iNOS in rat cardiomyocytes after reperfusion. In primary cultured cardiomyocytes, obestatin decreased LDH content and increased GSH level after I/R injury. Obestatin was also found to antagonize the apoptosis of cardiomyocytes in a dose-dependent manner. Western blot analysis showed that obestatin downregulated the expression of caspase-3 and Bax and upregulated the expression of Bcl-2. CONCLUSIONS Obestatin can protect cardiomyocyte from I/R-induced injury in vitro and in vivo. This beneficial effect is closely related with its properties of anti-inflammation, anti-cytotoxicity, and anti-apoptosis. The protective effect of obestatin might be associated with activation of Bcl-2 and inhibition of caspase-3 and Bax.

Clinical Outcomes of Coronary Artery Bypass Grafting vs Percutaneous Coronary Intervention in Octogenarians With Coronary Artery Disease.

BACKGROUND: The number of elderly people receiving treatment for coronary artery disease (CAD) is increasing, and there are few studies that compared the outcomes of coronary artery bypass grafting (CABG) and percutaneous coronary intervention (PCI) in the elderly. The objective of this study was to compare outcomes of CABG and PCI in octogenarians with CAD. METHODS: We conducted a search to identify articles that reported the results of 2-arm studies that compared CABG with PCI in octogenarians. The primary outcomes were short-term mortality and overall survival, and secondary outcomes included length of hospital stay and cerebrovascular accident (CVA) and myocardial infarction (MI) rates. RESULTS: Seven studies that enrolled 1879 patients who received CABG and 1432 treated with PCI were included. Short-term mortality was significantly less for patients in the PCI arms (odds ratio [OR], 1.47; 95% confidence interval [CI], 1.05-2.06; P = 0.02), as was duration of hospital stay (difference in means, 6.07; 95% CI, 2.81-9.34; P < 0.001). Patients in the CABG arms had longer overall survival (hazard ratio, 0.81; 95% CI, 0.73-0.89; P < 0.001). CVA and MI rates were similar (CVA: OR, 1.06; 95% CI, 0.57-1.95; P = 0.86; MI: OR, 0.70; 95% CI, 0.42-1.17; P = 0.17). CONCLUSIONS: The results suggest that physicians should consider not only the clinical features of CAD, but also the elderly patients future health outlook when choosing a revascularization procedure.

Apple Polyphenols Decrease Atherosclerosis and Hepatic Steatosis in ApoE-/- Mice through the ROS/MAPK/NF-κB Pathway.

In this study, we examined the effects of apple polyphenols (APs) on hyperlipidemia, atherosclerosis, hepatic steatosis and endothelial function and investigated the potential mechanisms. ApoE(-/-) mice were fed a western-type diet and orally treated with APs (100 mg/kg) or atorvastatin (10 mg/kg) for 12 weeks. Hyperlipidemia and atherosclerosis in the aortic sinuses and, and hepatic lipidosis were measured. The treatment with APs or atorvastatin induced a remarkable reduction in the atherosclerotic lesions and hepatic steatosis and decreased the levels of low-density lipoprotein, triglyceride, CCL-2 and VCAM-1 levels in the plasma. Conversely, the APs significantly increased the plasma levels of high-density lipoprotein (HDL) cholesterol and markedly up-regulated the glutathione peroxidase (GPx), catalase (CAT) and superoxide dismutase (SOD) levels in liver tissues. Moreover, the APs treatment modulated lipid metabolism by up-regulating the transcription of associated hepatic genes including PPARα, while down-regulating the transcription of SCAP and its downstream genes associated with lipid synthesis in the liver. Histological assessment showed that the APs treatment also reduced the macrophage infiltration in the aortic root plaque and the inflammatory cells infiltrations to the liver tissues. Moreover, we confirmed that the APs treatment greatly reduced the ox-LDL-induced endothelial dysfunction and monocyte adhesion to rat aortic endothelial cells (RAECs). Mechanistically, the APs treatment suppressed the ROS/MAPK/NF-κB signaling pathway, and consequently, reduced CCL-2, ICAM-1 and VCAM-1 expression. Our results suggest that the APs are a beneficial nutritional supplement for the attenuation of atherosclerosis.

[Plasma gamma-glutamyl transpeptidase level as a cardiovascular risk factor in elderly patients with hypertension or hypertension with diabetes mellitus].

OBJECTIVE: To investigate plasma gamma-glutamyl transpeptidase (γ-GGT) level as a cardiovascular risk factor in elderly patients with hypertension or hypertension with diabetes mellitus. METHODS: Forty-nine elderly patients of hypertension and 42 elderly patients of hypertension with diabetes mellitus and 39 healthy elderly subjects were enrolled in the study. The height, weight and blood pressure of patients were measured, serum C reactive protein and other biochemical indicators were detected. The relation between plasma γ-GGT and cardiovascular risk factors in three groups were analyzed. RESULTS: There was no significant difference in plasma γ-GGT levels among three groups. There was a positive correlation of plasma γ-GGT levels with systolic pressure, pulse pressure, hemoglobin A1c and CRP in control group. While in hypertension with diabetes mellitus group, plasma γ-GGT levels were correlated with systolic pressure, mean arterial pressure, fasting blood sugar and cystatin. CONCLUSION: Plasma γ-GGT might be a risk factor for cardiovascular diseases, and may be used as a predictive indicator for kidney injury in early patients with hypertension with diabetes mellitus.

Simvastatin protects human osteosarcoma cells from oxidative stress-induced apoptosis through mitochondrial-mediated signaling.

Apoptosis of osteoblasts has been proposed as the common basis of osteoporosis, with oxidative stress as the major cause. This study was performed to investigate the protective effect of simvastatin (0.001-0.1 µM) on 100 µM hydrogen peroxide (H2O2)-mediated oxidative stress-induced apoptosis in human osteosarcoma (MG63) cells and the molecular mechanisms involved. Cell apoptosis was evaluated by observation of morphological changes and Annexin V-propidium iodide double staining followed by flow cytometric analysis. MTS assays were used to evaluate cell viability. To investigate the underlying molecular mechanisms, the expression of caspase-3, caspase-9 and Bcl-2 were analyzed by Western blotting. Following stimulation with H2O2 for 24 h, a high proportion of MG63 cells underwent apoptosis, while a dose-dependent inhibition of apoptosis was observed in the presence of simvastatin. A significant, dose-dependent reduction in the expression of caspase-3 and caspase-9 protein induced by H2O2 in MG63 cells was observed in response to simvastatin and the Bcl-2 levels were increased. In conclusion, simvastatin protects MG63 cells from oxidative stress-induced apoptosis through downregulation of caspase-3 and caspase-9 activation and upregulation of Bcl-2 expression, suggesting a protective effect in osteoporosis.

Chewing substances with or without tobacco and risk of cardiovascular disease in Asia: a meta-analysis.

OBJECTIVE: To assess whether people who ever use any form of chewing substance in Asia are at increased risk of cardiovascular disease (CVD). METHODS: PubMed and ISI Web of Science were searched for relevant studies, with no limitation on language or study year. Studies were included if they provided quantitative estimate of the association between ever use of chewing substance and the occurrence of CVD. Two authors independently implemented inclusion criteria, abstracted study characteristics, and performed meta-analysis. Summary relative risks were estimated on the basis of a random effect model. We used Q statistic and Egger's test to examine heterogeneity across studies and potential publication bias, respectively. RESULTS: Eight eligible studies were included. The relative risk of CVD for ever using chewing substances with or without tobacco was 1.26 (95% confidence interval (CI) 1.12-1.40), which was unchanged when restricted to cohort studies [1.25 (1.08-1.42)] or cohort studies in Taiwan [1.31 (1.12-1.51)]. The summary relative risk for ischemic heart disease was 1.27 (1.02-1.52), and was lowered to 1.26 (0.85-1.67) after exclusion of a cross-sectional study. The overall relative risk for cerebrovascular disease was 1.32 (1.08-1.56). On the basis of the Taiwan data, the summary relative risk of CVD for betel (Areca catechu) chewing was 1.30 (1.17-1.44). Data on dose-response were limited to betel chewing in Taiwan, suggesting a relationship between risk of CVD and cumulative exposure. Two large cohorts in Taiwan reported a greater risk of CVD with betel chewing than with smoking. CONCLUSIONS: An association was detected between betel chewing with or without tobacco and the risk of CVD. Betel chewing may impose a greater CVD risk than smoking. More effort is needed in developing betel chewing cessation programmes. The relationship between betel chewing and subgroups of CVD requires further investigation.

Construction of a recombinant adenovirus vector expressing IL-18BP/IL-4 fusion gene and the anti-inflammatory effect induced by this gene on lipopolysaccharide-stimulated synovial fibroblasts.

OBJECTIVE: To construct a recombinant adenovirus vector for expressing the IL-18 binding protein (IL-18BP)/IL-4 fusion gene and confirm the anti-inflammatory effect of this gene. MATERIALS AND METHODS: The recombinant virus expressing IL-18BP/IL-4 fusion protein (AD-IL-18BP/IL-4) was constructed. AD-IL-18BP/IL-4 was used to infect synovial fibroblasts (SF). ELISA and Western blot analysis were used to determine the expressions of the proteins IL-4 and IL-18BP. To investigate the protective effects of this vector on rheumatoid arthritis, SF were infected with AD-IL-18BP/IL-4 and stimulated by LPS (1 microg/ml) 4 h later. The expression levels of TNF-alpha, IL-6, IL-8, and IL-18 in the culture supernatant were detected by ELISA and production of PGE2 and NO was estimated. The protein expression of COX-2, iNOS, and NF-kappaB p50 in treated SF was analyzed by Western blot. RESULTS: AD-IL-18BP/IL-4 can effectively express the IL-18BP/IL-4 fusion protein. The expressions of TNF-alpha, IL-6, IL-8, and IL-18 were significantly inhibited in LPS-stimulated SF after treatment with AD-IL-18BP/IL-4. The production of PGE2 and NO was significantly decreased. Moreover, NF-kappaB p50, COX-2, and iNOS levels in SF were markedly suppressed by AD-IL-18BP/IL-4. CONCLUSION: AD-IL-18BP/IL-4 can suppress the production and expression of inflammatory cytokines such as COX-2, iNOS, and NF-kappaB in LPS-stimulated SF.

Resistin does not down-regulate the transcription of insulin receptor promoter.

OBJECTIVE: To detect the effect of resistin on the transcription of insulin receptor promoter. METHODS: Luciferase reporter gene was fused downstream of human insulin receptor promoter and the enzymatic activity of luciferase was determined in the presence or absence of resistin. The resistin expressed with plasmid was stained with antibody against Myc tag which was in frame fused with resistin coding sequence, and then imaged with confocal microscopy. RESULTS: The treatment of pIRP-LUC transfected cells with recombinant resistin did not result in significant difference in the enzymatic activity of luciferase compared to the untreated cells. Cell staining showed that green fluorescence could be observed in the cytoplasm, but not in the nucleus. CONCLUSION: The results suggest that the endogenous resistin may functionally locate in the cytoplasm, but does not enter the nucleus and not down-regulate the transcription of insulin receptor promoter.

Study of Resistin gene expression in peripheral blood mononuclear cell and its gene polymorphism in a small range population.

OBJECTIVE: To observe the expression of Resistin mRNA in peripheral blood mononuclear cells and its gene polymorphism in coding region in a small range population in Zhejiang Province of China. METHODS: Eighty-three cases of type 2 diabetes mellitus and 53 healthy people were included. The expression of Resistin mRNA in peripheral blood mononuclear cells was detected by RT-PCR and semi-quantitative PCR assay. The sequencing work was done in Resistin cDNA and gene polymorphism was analyzed. RESULTS: At the same condition, in 83 diabetes patients, Resistin mRNA was detected in 23 cases (11 males and 12 females). There was no Resistin mRNA expression in 53 healthy people. The ratio of PCR products between Resistin and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) was from 0.564 to 1.238, averaging 0.804+/-0.436. The sequence of Resistin cDNA is almost identical with each other and with that in GenBank with no single nucleotide polymorphism being found. CONCLUSION: Resistin mRNA is expressed in human peripheral blood mononuclear cells in some type 2 diabetes mellitus, but its expression is at a low level. Among the experiment population we did not find polymorphism phenomenon in Resistin coding region. The different individual's Resistin coding region is highly coincident.

Relationship between resistin level in serum and acute coronary syndrome or stable angina pectoris.

OBJECTIVE: To investigate the relationship between serum resistin level and acute coronary syndrome (ACS) or stable angina pectoris (SAP). METHODS: Sixty-five patients, with coronary artery disease, were enrolled and divided into three subgroups: acute myocardial infarction (AMI), unstable angina pectoris (UAP) and SAP, and 26 healthy people were recruited as controls in the cross-sectional study. Serum resistin levels were determined by ELISA (enzyme-linked immunosorbent assay), and WBC (white blood cell count), hsCRP (high sensitive C-reaction protein), CK(max) (maximum of creatinkinase), CK-MB(max) (maximum of isozyme of creatinkinase) and cTnI(max) (maximum of troponin) were measured by standard laboratory methods. RESULTS: The serum resistin levels were 4 folds higher in AMI patients, 2.43 folds in UAP patients and 1.12 folds in SAP patients than in the healthy controls (P<0.05). The resistin levels were also significantly different between AMI [(8.16+/-0.79) ng/ml], UAP [(5.59+/-0.75) ng/ml] and SAP [(3.45+/-0.56) ng/ml] groups (P<0.01); WBC, hsCRP, CK(max), CK-MB(max) and cTnI(max) were significantly increased in AMI patients over UAP and SAP patients. Spearman analysis showed that serum resistin levels were positively correlated with WBC (r=0.412, P=0.046), hsCRP (r=0.427, P=0.037), CK(max), CK-MB(max) and cTnI(max) (r=0.731, 0.678, 0.656; P<0.01). CONCLUSION: Serum resistin levels increased with inflammatory factors and myocardial impairment. The results suggest that human resistin might play an important role in the pathogenesis of atherosclerosis and AMI as an inflammatory factor.

[Atherosclerosis and androgen levels in elderly males].

OBJECTIVE: To observe the relation of androgen levels and atherosclerosis (AS) in elderly males. METHODS: Both carotid arteries and arteries of lower extremity were examined with Doppler ultrasonography. Those with arteriosclerosis and much atheromatous plaque were designated as case group, and those with normal results formed control group. Total testosterone (TT), free testosterone (FT) and estradiol (E2) were measure by radioimmunoassay, HDL-C, LDL-C, TC and TG were assayed by colorimetry, vascular endothelium growth factor (VEGF), tumor necrosis factor-alpha (TNF-alpha), interleukin-6 (IL-6), intercellular adhesion molecule (sICAM-1) were determined by ELISA. RESULT: FT was significantly lower in case group than in control group (P<0.01), no differences were found in TT, E2. HDL-C in control group was higher than that in case group (P<0.01), TC and TG were higher in case group than those in control group (P<0.05). HDL-C was correlated positively and LDL-C was negatively with FT level, while both TC and TG in case group had negative relation with FT. VEGF was higher in case group (P<0.05), and it had negative relation with FT in both groups. TNF-alpha and IL-6 were significantly higher in case group (P<0.05), and they had negative relation with FT. sICAM-1 was significantly lower in control group than it in case group (P<0.01). CONCLUSION: The normal androgen levels, especially FT, have beneficial effect in AS development in elderly males. Low FT level may be an independent risk factor in AS development.

Study of androgen and atherosclerosis in old-age male.

OBJECTIVE: To observe the difference of androgen and inflammatory cytokines level in atherosclerosis and analyse their relations. METHOD: Both carotid arteries and arteries of lower extremity were subjected to ultrasonic examination by Doppler's method. Those with much atheromatous plaque formation were ranged into case group, and those with normal result formed control group. Total, free testosterone and estradiol were assayed by radioimmunoassay. C reactive protein (CRP) was assayed by nepheloturbidity. Tumor necrosis factor-alpha (TNF-alpha), Interleukin-6 (IL-6), Interleukin-8 (IL-8), Interleukin-10 (IL-10), Interleukin-18 (IL-18), soluble intercellular adhesion molecule-1 (sICAM-1) and soluble vascular cell adhesion molecule-1 (sVCAM-1) were assayed by ELISA. The mean difference between two groups and the correlation between free testosterone and cytokines were analysed. RESULTS: Free testosterone was (6.337+/-3.371) pg/L in case group and (11.375+/-4.733) pg/L in control group, P<0.01. No differences were found in total testosterone and estradiol. CRP was (27.294+/-10.238) mg/L in case group and (12.843+/-6.318) mg/L in control group, P<0.01. IL-6 was (41.700+/-31.385) pg/L in case group and (25.396+/-20.772) pg/L in control group, P<0.05. IL-8 was (89.249+/-58.357) pg/L in case group and (67.873+/-31.227) pg/L in control group, P<0.05. sICAM-1 was (470.491+/-134.078) pg/L in case group and (368.487+/-97.183) pg/L in control group, P<0.01. sVCAM-1 was (537.808+/-213.172) pg/L in case group and (457.275+/-157.273) pg/L in control group, P<0.05. There were no differences in TNF-alpha, IL-10 and IL-18. Correlation analysis showed that FT (free testosterone) had negative correlation with CRP, IL-6 and sICAM-1. Among them FT had well correlation with CRP, correlation index was -0.678. CONCLUSION: Free testosterone was in negative correlation with atherosclerosis in old-age male. Free testosterone may have the role of anti-atherosclerosis, and this effect was not achieved by its transformation to estradiol. Low free testosterone level was followed by increased level of inflammatory cytokines. Low free testosterones coexist with inflammation and they both affect the process of atherosclerosis in old-age male.