Correlations Among Maternal and Infant Factors, Lead Exposure, and Serum Prolactin Levels During Lactation: A Cross-sectional Study in Indonesia.

OBJECTIVES: Prolactin is vital for breastfeeding and milk production, and its secretion is influenced by factors related to the mother, infant, and environment. To date, no study has concurrently investigated the correlation of these factors with serum prolactin levels during lactation. Therefore, the objective of this study was to investigate the correlations among maternal and infant factors, lead exposure, and serum prolactin levels during lactation. METHODS: A cross-sectional approach was employed in Surabaya, Indonesia, among 110 exclusively lactating mothers. The mothers' daily diets were determined using multiple 24-hour recalls, while blood lead levels were measured with inductively coupled plasma mass spectrometry. Serum prolactin levels were assessed using the electrochemiluminescence immunoassay. For bivariate analysis, we employed the Spearman correlation, Mann-Whitney, and Kruskal-Wallis tests, while for multivariate analysis, we utilized multiple linear regression. RESULTS: The average serum prolactin level of the lactating mothers was 129.19±88.96 ng/mL. Positive correlations were found between serum prolactin levels and breastfeeding frequency (p < 0.001), protein intake (p < 0.001), and calcium intake (p = 0.011) but had negative correlation with blood lead levels (p < 0.001) and vitamin B6 intake (p = 0.003). Additionally, prolactin levels were not significantly associated with maternal age; parity; intake of calories, vitamin D, vitamin E, zinc, folic acid, magnesium, or iron; infant age; or infant sex. CONCLUSIONS: Breastfeeding frequency had a stronger positive relationship with serum prolactin levels than protein and calcium intake. However, lead exposure was associated with reduced serum prolactin levels during lactation. Consequently, specific interventions from policymakers are necessary to manage breastfeeding in mothers exposed to lead.

The Role of Protein Intake on the Total Milk Protein in Lead-Exposed Lactating Mothers.

Protein is an essential macronutrient for the growth and development of infants. Protein levels in lactating mothers are dynamic and influenced by various factors, particularly the environment and maternal characteristics. Therefore, this study aimed to evaluate the complex correlation between maternal blood lead levels (BLLs), maternal diet, and total milk protein. The Kruskal-Wallis test was used to compare total milk protein in the three groups of lead exposure, while Spearman's correlation was used to assess the correlation between maternal diet, BLLs, and total milk protein. The multivariate analysis used multiple linear regression. The results showed that the median of maternal BLLs and total milk protein were 3.3 µg/dL and 1.07 g/dL, respectively. Maternal protein intake and current BMI had a positive correlation with total milk protein, while BLLs had a negative correlation. BLLs ≥ 5 µg/dL had the most significant impact on reducing the total milk protein (p = 0.032). However, increasing maternal protein intake can effectively maintain total milk protein levels in mothers with BLLs under 5 µg/dL (p < 0.001). It is crucial to measure BLLs in lactating mothers residing in areas exposed to lead because high maternal protein intake can only maintain total milk protein levels when the BLLs are <5 µg/dL.

Metabolite profiling, hypolipidemic, and anti-atherosclerosis activity of mixed vegetable fermentation extract.

Although positive association between fermented vegetables intake with the risk of coronary heart disease (CHD) has increased attention nowadays, the metabolite profiling and the mechanism of action are still elusive. This study designed to investigate the secondary metabolites, hypolipidemic, and anti-atherogenic effect of mixed vegetable fermentation extract (MVFE). The metabolite screening of the MVFE was assessed using the Liquid Chromatography Tandem Mass Spectrophotometer (LC-MS/MS) method. The result of LC-MS/MS was used as ligands to inhibit the binding of oxidized LDL (oxLDL) and Cluster Differentiation 36 (CD36), Scavenger Receptor A1 (SRA1), Lectin-type oxidized LDL receptor 1 (LOX1). This work was performed with molecular docking using Discovery Studio 2021, PyRx 0.9, and Autodock Vina 4.2 followed by analyzing Network Pharmacology, Protein Protein Interaction (PPI) using Cytoscape 3.9.1 and String 2.0.0. Finally, the clinical effect of MVFE was evaluated using in vivo study. Twenty rabbits were assigned to normal, negative control, and MVFE group that were fed with standard diet, high fat diet (HFD), HFD supplemented with MVFE 100, 200 mg/kg BW, respectively. The serum level of Total Cholesterol (TC) and Low-Density Lipoprotein (LDL-c) were detected at the end of week 4. The LC-MS/MS analysis identified 17 compounds categorized as peptides, fatty acids, polysaccharides, nucleoside, flavonoids, flavanols, and phenolic compounds. Based on the docking study, more negative binding affinity was observed in the interaction between metabolites with the scavenger receptors (SR) than simvastatin. The number of nodes and edges based on Network Pharmacology analysis were 268 and 482, respectively. The PPI network showed that MVFE metabolites exerts its athero-protective effect by modulating various cellular processes including inflammation, improvement of endothelial function, and modulation of lipid metabolism. Blood TC and LDL-c concentrations in the negative control (458.82 ± 82.03; 191.87 ± 92.16 mg/dL) were higher significantly compared to the normal group (87.03 ± 29.27; 43.33 ± 5.75 mg/dL). The MVFE administration decreased the TC (100, 200 mg/kg BW MVFE: 269.96 ± 85.34; 130.17 ± 45.02 mg/dL) and LDL-c level (100, 200 mg/kg BW MVFE = 87.24 ± 22.85; 41.82 ± 11.08 mg/dL) dose-dependently (p < 0,001). The secondary metabolites derived from fermented mixed vegetables extract might be developed as a potential strategy to prevent CHD by targeting the multiple pathways in atherosclerosis.

Curcuma longa supplementation reduces MDA, TNF-α, and IL-6 levels in a rat model exposed to soot particulates.

Background: Particulate matter (PM) is one of the important components in air pollution that can cause endothelial vascular dysfunction through exacerbation of atherosclerosis and inflammation of the respiratory system. Increased levels of malondialdehyde (MDA) in blood plasma can be an indicator of oxidative stress. Then, macrophages can secrete proinflammatory cytokines that will stimulate immune cells and vascular endothelial cells to release inflammatory cytokines, such as interleukin-6 (IL-6) and tumor necrotic factor-α (TNF-α). Curcuma longa works by scavenging the active free radicals involved in the peroxidation process. Aims: This study aims to prove that the administration of C. longa can reduce MDA, TNF-α, and IL-6 levels in Rattus norvegicus exposed to soot particulates. Methods: The subjects of this study were 30 male rats which were divided into 5 treatment groups with the following: (C0): negative control; (C+): positive control; (T1): Treatment group 2, rats exposed to particulate soot at a concentration of 1,064 mg/m3 for 8 hours and given C. longa at a dose of 1 mg/kg bw; (T2): Treatment group 3 was rats exposed to soot particulates at a concentration of 1,064 mg/m3 for 8 hours and given C. longa at a dose of 2 mg/kg bw; (T3): Treatment group 4 was rats exposed to soot particulates at a concentration of 1,064 mg/m3 for 8 hours and given C. longa at a dose of 3 mg/kg bw.Giving the C. longa extract orally with a probe every day for 30 days after treatment of exposure to soot. Examination of MDA, TNF-, and IL-6 levels with the ELISA method. Results: The administration of C. longa can reduce MDA while the lowest MDA levels were obtained in the T3 treatment with an average of 1.542 ± 0.231. The results of the description of the lowest levels of TNF-α were obtained in the C-treatment with an average of 55.981 ± 4.689. Then, the lowest levels of IL-6 were obtained in the C-treatment with an average of 2.292 ± 0.461. Conclusion: The results stated that the administration of C. longa could reduce MDA levels, TNF-α, and IL-6 levels. Curcuma longa as an anti-inflammatory and anti-oxidant play an effective role in inhibiting inflammation by decreasing IL-6 cytokine and TNF-α. Curcuma longa can inhibit lipid peroxidation initiated by free radicals and then reduce MDA levels.

Anti-atherosclerotic activity of aqueous extract of Ipomoea batatas (L.) leaves in high-fat diet-induced atherosclerosis model rats.

OBJECTIVES: Cardiovascular diseases, especially atherosclerosis, are the leading cause of human mortality in Indonesia. Ipomoea batatas (L.) is a food plant used in Indonesian traditional medicine to treat cardiovascular diseases and related conditions. We assessed the anti-atherosclerotic activity of the aqueous extract of I. batatas leaves in a rat model of high-fat diet-induced atherosclerosis and its mechanism. METHODS: The presence of amino acid content in the I. batatas L. purple variant was determined by liquid chromatography high-resolution mass spectrometry (LC-HRMS). Thirty male Wistar rats were divided into five groups (n=6/group), i.e., standard diet group (SD), high-fat diet group (HF), and HF plus I. batatas L. extracts orally (625; 1,250; or 2,500 mg/kg) groups. The numbers of macrophages and aortic wall thickness were analyzed histologically. Immunohistochemical analyses were performed to assess foam cells-oxidized low-density lipoprotein (oxLDL), endothelial nitric oxide synthase (eNOS), and vascular endothelial growth factor (VEGF) expression in the aorta. RESULTS: LC-HRMS analysis showed nine amino acid content were identified from I. batatas L. In vivo study revealed that oral administration of I. batatas L. leaf extract alleviated foam cells-oxLDL formation and aortic wall thickness caused by high-fat diet atherosclerosis rats. Further, I. batatas L. leaf extract promoted the number of macrophages and modulated VEGF and eNOS expression in the aorta. CONCLUSIONS: I. batatas L. leaf extract shows a positive anti-atherosclerosis effect. Furthermore, the mechanism may promote the macrophages, eNOS, VEGF expressions, and inhibition of foam cells-oxLDL formation and aortic wall thickness with the best dosage at 2,500 mg/kg. This could represent a novel approach to prevent cardiovascular diseases.

The role of methylenetetrahydrofolate reductase C677T gene polymorphism as a risk factor for coronary artery disease: a cross-sectional study in the Sidoarjo Regional General Hospital.

Introduction: hyperhomocysteinemia (HHcy) may contribute to an increased risk of coronary artery disease (CAD). The underlying mechanisms are not well understood, but other than dietary intake factors, hyperhomocysteinemia may genetically result from a methylenetetrahydrofolate reductase (MTHFR) C677T gene polymorphism. A cross-sectional study was performed to assess whether this mutation was a potential genetic risk factor for CAD. Methods: this cross-sectional study was performed on 30 CAD patients and 30 normal healthy controls at Sidoarjo Regional General Hospital. The polymorphisms of the MTHFR C677T gene was assessed by polymerase chain reaction (PCR), and plasma homocysteine was measured by chemiluminescence immunoassay (CLIA) and then compared between CAD patients and control subjects by the multivariate logistical regression model. Results: results from an independent sample t-test analysis showed that plasma homocysteine concentrations were significantly higher in CAD patients compared to the control group individuals (13.91 ± 4.55 µmol/L vs 10.97 ± 3.45 µmol/L; p<0.05). There were no significant correlations between MTHFR C677T gene polymorphism and other risk factors, such as age at diagnosis with acute coronary syndrome, sex, smoking, lipid profile, diabetes, hypertension, C-reactive protein (CRP), creatinine, and homocysteine (p>0.05). In multivariate analysis models, the C677T genotype frequencies were insignificantly different between CAD patients and control subjects (p>0.05). Meanwhile, the results of adjusted odds ratio (aOR), 95% confidence interval (CI), and p-value for homocysteine, age, and smoking were aOR: 1.264, 95% CI : 1.042-1.535, p = 0.018; aOR: 0.916, 95% CI: 0.842-0.997, p = 0.043, and aOR: 5.428, 95% CI 1.532-19.226, p = 0.009, respectively. Homocysteine, age, and smoking were significantly different between CAD patients and control subjects (p<0.05). Conclusion: hyperhomocysteinemiais significantly correlated with an increased risk of CAD, but MTHFR C677T gene polymorphism might not contribute to increased CAD risk.

miR-155-5p predictive role to decelerate foam cell atherosclerosis through CD36, VAV3, and SOCS1 pathway.

MicroRNAs (miRNAs) are noncoding RNA molecules that play a significant role in atherosclerosis pathogenesis through post-transcriptional regulation. In the present work, a bioinformatic analysis using TargetScan and miRdB databases was performed to identify the miRNAs targeting three genes involved in foam cell atherosclerosis (CD36, Vav3, and SOCS1). A total number of three hundred and sixty-seven miRNAs were recognized and only miR-155-5p was selected for further evaluation based on Venn analysis. Another objective of this study was to evaluate the biological process and regulatory network of miR-155-5p associated with foam cell atherosclerosis using DIANA, DAVID, Cytoscape, and STRING tools. Additionally, the comprehensive literature review was performed to prove the miR-155-5p function in foam cell atherosclerosis. miR-155-5p might be related with ox-LDL uptake and endocytosis in macrophage cell by targeting CD36 and Vav3 genes which was showed from the KEGG pathways hsa04979, hsa04666, hsa04145 H, hsa04810, and GO:0099632, GO:0060100, GO:0010743, GO:001745. Furthermore, miR-155-5p was also predicted to increase the cholesterol efflux from macrophage by inhibit SOCS1 expression based on KEGG pathway hsa04120. Eleven original studies were included in the review and strongly suggest the role of miR-155-5p in foam cell atherosclerosis inhibition.

The significance of Agaricus blazei as an immunomodulator of the level of IL-17 in Balb/C mice with atherosclerosis.

Atherosclerosis is a disease caused by an inflammatory response which involved the interaction between endothelial cells, macrophages and lymphocytes, and is closely related to IL-17 regulation. This study is important to investigate the activity of Agaricusblazei in modulating the immunological activity based on the profile of CD4+IL-17+, CD8+IL-17+, and CD11b+ IL-17+ in high-fat diet (HFD)-induced Balb/c mice. Mice in dietary groups were fed with HFD and then fed with A. blazei extract with three different doses including D1 (100 mg/kg BW), D2 (200 mg/kg BW), and D3 (400 mg/kg BW) once a day for 12 weeks. The cells were analyzed using flow cytometry and tested statistically with one-way ANOVA with α = 0.05 by using SPSS 16.00 software. The results showed that mice with HFD treatment had a higher level of Lp-PLA2 (atherosclerosis marker) compared with the control group (data not shown). The level of IL-17 in the atherosclerotic mice in the D1 group was significantly depleted compared to the control group. Of the three doses above, D1 may be an optimal dose to minimize or prevent the damage from atherosclerosis than the other doses.

Polysaccharide peptide (PsP) Ganoderma lucidum: a potential inducer for vascular repair in type 2 diabetes mellitus model.

INTRODUCTION: The increasing blood glucose level due to insulin resistance which occurs in diabetes mellitus (DM) may cause vascular damage. This study aims to prove the effect of the polysaccharide peptide (PsP) Ganoderma lucidum on improving vascular damage through an increase of circulating endothelial cells and circulating endothelial cells (CEC) ratio, decreased H2O2, triglyceride (TG), total cholesterol (TC) and insulin resistance in type 2 DM. METHODS: Our study is a true experimental study with randomized posttest control group design that used 35 Wistar rats divided into five groups: normal, control (+) and three groups of different variant PsP doses 50, 150 and 300 mg/kg BW (n=7). RESULTS: By using one-way ANOVA and post-hoc Duncan test, the results show a significant increase of endothelial progenitor cell (EPC) concentration (p=0.000) and ratio EPC:CEC (0.000) by dose-dependent fashion and also reduced CEC concentration (p=0.001), H2O2 (p=0.03), TG (p=0.001), TC (p=0.01) and insulin resistance (p=0.003). CONCLUSION: In this study, PsP induced endothelial repairing process and reduced the risk factor with 300 mg/kg BW as optimum dose. However, further research on EPC and CEC detection markers is important. Further research on PsP and clinical trial for commercial uses is also needed.

The role of polysaccharide peptide of Ganoderma lucidum as a potent antioxidant against atherosclerosis in high risk and stable angina patients.

OBJECTIVES: Antioxidants can reduce oxidative radicals that affect the early phase of atherogenesis, that is endothelial dysfunction. Polysaccharide Peptide (PsP) derived from Ganoderma lucidum has an active substance in the form of ß-glucan. Previous studies have proven the PsP of Ganoderma lucidum as an effective antioxidant in atherosclerotic rats and shows no toxicity in animal model. This study aims to prove the effect of PsP as potent antioxidant in high risk and stable angina patients. METHOD: This is a clinical trial conducted to 37 high risk and 34 stable angina patients, which were determined based on ESC Stable CAD Guidelines and Framingham risk score, with pre and post test design without control group. The parameters are superoxide dimustase (SOD) and malondialdehyde (MDA) concentration, circulating endothelial cell (CEC) and endothelial progenitor cell (EPC) counts. The patients were given PsP 750mg/day in 3 divided dose for 90days. Paired t-test was performed for normally distributed data, and Wilcoxon test for not normally distributed data, and significant level of p≤0,05. RESULTS: SOD level in high risk patients slightly increased but not statistically significant with p=0,22. Level of SOD in stable angina group significantly increased with p=0,001. MDA concentration significantly reduced in high risk and stable angina patients with p=0.000. CEC significantly reduced both in high risk and stable angina patients, with p=0.000 in both groups. EPC count significantly reduced in high risk and stable angina with p=0.000. CONCLUSION: PsP of Ganoderma lucidum is a potent antioxidant against pathogenesis of atherosclerosis in stable angina and high risk patients.

The reduction of aorta histopathological images through inhibition of reactive oxygen species formation in hypercholesterolemia rattus norvegicus treated with polysaccharide peptide of Ganoderma lucidum.

OBJECTIVES: Atherosclerosis is chronic inflammatory process triggered by oxidative stress. Oxidative stress can increase hydrogen peroxide (H2O2) level, which induce atherosclerosis through the processes such as formation of perivascular adipose tissue (PVAT), foam cells, and atherosclerotic plaque. Antioxidant is needed to control negative effects of oxidative stress. One source of antioxidant, which has potential to be developed, is PsP from Ganoderma lucidum. This study aims to prove the effect of PsP in decreasing H2O2, PVAT, foam cells and atherosclerotic plaque. MATERIALS AND METHODS: This study was experimental randomized post-test with control group design using 25 Rattus norvegicus Wistar strain rats. Rats were divided into 5 groups (negative control, positive control, and 3 high-fat diet group with PsP dose: 50, 150, 300 mg/kgBW). Measured parameters were H2O2, PVAT, foam cell, and atherosclerotic plaques. Analysis of variance (ANOVA) was used for statistical analysis, followed by post hoc test. RESULTS: Mean H2O2 levels, PVAT thickness, foam cell numbers, and atherosclerotic plaque were low in negative control group. ANOVA showed that PsP significantly (P<0.05) reduced H2O2 levels, PVAT thickness, foam cells numbers and atherosclerotic plaque width. CONCLUSION: PsP dose of 300 mg/kgBW has the most significant effect in decreasing H2O2 levels, PVAT thickness, number of foam cells, and atherosclerotic plaque width. Based on the results of this research, PsP can be recommended as antioxidant to control pathogenesis of atherosclerosis.

Endothelial progenitor cells proliferated via MEK-dependent p42 MAPK signaling pathway.

Endothelial progenitor cells (EPCs) clinical applications have been well reported. However, due to low number of EPCs that could be isolated, EPCs expansion study became one of the main focuses. Some optimized mediums to culture EPCs were currently available. However, the proliferation signaling pathway is not clearly disclosed yet. Peripheral blood was collected from eight healthy subjects, followed by mononuclear cells (MNCs) isolation. MNCs were then prepared and cultured for 2 days. After that, non-adherent cells were harvested and further cultured for 3 days. Resulted colony-forming unit (CFU)-Hill colonies were documented and enumerated under an inverted light microscope. To detect membrane markers, immunofluorescence was performed to detect CD34, VEGFR-2, and CD133. Cell documentation was conducted under a fluorescence microscope. To check cell proliferation, XTT Cell Proliferation Assay Kit was used according to kit insert. To detect possible activation of p44/42 MAPK, western blot was performed to detect p44/42 MAPK and phosphorylated p44/42 MAPK. All visualized bands were captured and quantified. Our results showed that EPCs markers (CD34, CD133 and VEGFR-2) were detected in 3 days culture. From XTT cell proliferation assay and CFU enumeration results, we found that EPCs proliferated significantly (p = 0.012) with addition of supplement. Phosphorylated-p42 MAPK expression of EPCs treated with supplement was significantly higher than the one of EPCs without treatment. Significant inhibition of p42 MAPK phosphorylation by U0126 was observed (p = 0.012). By pretreatment of U0126, number of viable cells and CFUs treated with supplement was significantly decreased (p = 0.012). Our results showed that MEK-dependent p42 MAPK pathway might play an important role in EPCs proliferation.

Effect of Scurrula atropurpurea on nitric oxide, endothelial damage, and endothelial progenitor cells of DOCA-salt hypertensive rats.

OBJECTIVES: To know whether Scurrula atropurpurea is able to modulate total plasma nitrate/nitrite levels, decrease endothelial damage, and increase endothelial progenitor cells (EPCs) in hypertensive rats. MATERIALS AND METHODS: The rats were divided in 5 groups: control (normotensive) group, Desoxy cortico sterone (DOCA)-salt hypertensive group, and three DOCA-salt hypertensive groups. All 5 groups received methanolic extract of S. atropurpurea (MESA) at a dosage of 50; 100; and 200 mg/KgBW. Serum nitric oxide (NO) was assayed by colorimetric. Circulating endothelial cells (CECs) and EPCs were assayed using flow cytometry. RESULTS: The administration of MESA100 and MESA200 elevated the total plasma nitrate/nitrite levels but cannot reach the level in control group. MESA100 and MESA200 also elevated the EPCs number compared with hypertensive group. The administration of MESA significantly (P< 0.05) decreased the CECs number compared to hypertensive groups. CONCLUSION: Methanolic extract of S. atropurpurea is able to modulate total plasma nitrate/nitrite levels and diminish endothelial damage via increasing EPCs.

Vasa vasorum anti-angiogenesis through H2O2, HIF-1α, NF-κB, and iNOS inhibition by mangosteen pericarp ethanolic extract (Garcinia mangostana Linn) in hypercholesterol-diet-given Rattus norvegicus Wistar strain.

BACKGROUND: Oxidative stress in atherosclerosis produces H2O2 and triggers the activation of nuclear factor kappa beta (NF-κB) and increase of inducible nitric oxide synthase (iNOS). The formation of vasa vasorum occurs in atherosclerosis. Vasa vasorum angiogenesis is mediated by VEGFR-1 and upregulated by hypoxia-inducible factor-1α (HIF-1α). The newly formed vasa vasorum are fragile and immature and thus increase plaque instability. It is necessary to control vasa vasorum angiogenesis by using mangosteen pericarp antioxidant. This study aims to demonstrate that mangosteen pericarp ethanolic extract can act as vasa vasorum anti-angiogenesis through H2O2, HIF-1α, NF-κB, and iNOS inhibition in rats given a hypercholesterol diet. METHODS: This was a true experimental laboratory, in vivo posttest with control group design, with 20 Rattus norvegicus Wistar strain rats divided into five groups (normal group, hypercholesterol group, and hypercholesterol groups with certain doses of mangosteen pericarp ethanolic extract: 200, 400, and 800 mg/kg body weight). The parameters of this study were H2O2 measured by using colorimetric analysis, as well as NF-κB, iNOS, and HIF-1α, which were measured by using immunofluorescence double staining and observed with a confocal laser scanning microscope in aortic smooth muscle cell. The angiogenesis of vasa vasorum was quantified from VEGFR-1 level in aortic tissue and confirmed with hematoxylin and eosin staining. RESULTS: Analysis of variance test and Pearson's correlation coefficient showed mangosteen pericarp ethanolic extract had a significant effect (P<0.05) in decreasing vasa vasorum angiogenesis through H2O2, HIF-1α, NF-κB, and iNOS inhibition in hypercholesterol-diet-given R. norvegicus Wistar strain. CONCLUSION: Mangosteen pericarp ethanolic extract 800 mg/kg body weight is proven to decrease vasa vasorum angiogenesis. Similar studies with other inflammatory parameters are encouraged to clarify the mechanism of vasa vasorum angiogenesis inhibition by mangosteen pericarp ethanolic extract.

Level of circulating endothelial cells and expression of nuclear factor kappa beta of human's peripheral blood mononuclear cells in subjects with certain conditions.

AIM: to detect the levels of CEC and expression of NFκB in the three groups of subjects with certain conditions. METHODS: this study is an exploratory study using human peripheral blood samples. The study subjects comprised three groups, the group of 23 healthy people, a group of 35 people that has one or more risk factors for vascular disease and a group of 15 vascular disease patients (coronary heart disease, diabetes mellitus, stroke). CEC were isolated from peripheral blood mononuclear cells (PBMC). CEC level is identified through the measurement of CD45 and CD146 by flowcytometry method. NFκB expression is recognized by ELISA method (imgenex, USA). RESULTS: the highest average levels of CEC were found in the sick group (28.6%). The highest average expression of NFκB (924.9495) is found in the group with risk factors. The lowest average expression of NFκB and CEC is found in the healthy group. Statistical analysis of ANOVA at the interval confidence of 95% shows a significant difference (p=0.00) levels of CEC and NFκB expression between the healthy group with the group with the risk of cardiovascular disease (CVD) and patients with known CVD. CONCLUSION: increase of level CEC and NFκB expression has a strong relationship with vascular disease and its risk factors.

Green tea polyphenols inhibit oxidized LDL-induced NF-KB activation in human umbilical vein endothelial cells.

AIM: To examine the anti-inflammatory effects of green tea polyphenols on oxLDL-mediated TNFalpha expression and NF-KB activation in the human umbilical vein endothelial cells (HUVECs). METHODS: We postulate that green tea polyphenols regulate TNF-alpha gene expression by modulating NF-KB activation through their inhibition effect on IKB Kinase (IKK) activity and as scavenger of free radicals. Pretreatment of green tea polyphenols reduced oxLDL-induced production of proinflammatory cytokine TNF-alpha and NF-KB activation in dose dependent manner (p < 0.05). Post hoc comparison test with Mann Whitney between various dosage of green tea polyphenols in inhibition of NF-KB activation showed significant result (p < 0.05). RESULTS: In TNF-alpha expression, there was also declined TNF-alpha productions (p 0.09; 0.2 vs 0.4mg/ml: ns). The effect of green tea polyphenols on TNF-alpha expression were determined by Mann-Whitney test. There is significant difference between the first dose (0.1mg/ml) vs 0.2mg/ml polyphenols (p=0.009); between 0.1 vs 0.4 mg/ml polyphenols (p=0.009). There was no difference when the dose was increased from 0.2 mg/ml to 0,4 mg/ml polyphenols (0.141). In this study, green tea polyphenols showed significant effects on the inhibition of TNF-alpha through NF-KB activation pathway in HUVECs with oxidized LDL. CONCLUSION: Green tea polyphenol can be used to prevent the development of atherosclerosis.