The Indonesian Biomedical Journal

18-36

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: The prevailing view that eukaryotic cells are restrained from intercellular exchange of genetic information has been challenged by recent reports on nanotubes, exosomes, apoptotic bodies, and nucleic acid—binding peptides that provide novel pathways for cell—cell communication, with implications in health and disease. CONTENT: Microparticles (MPs) are a heterogeneous population of small plasma membrane structures that serve as important signaling structures between cells. MPs are composed of a phospholipid bilayer that exposes transmembrane proteins and receptors and encloses cytosolic components such as enzymes, transcription factors, and mRNA derived from their parent cells. Growing evidence suggests that MPs regulate inflammation, stimulate coagulation, affect vascular functions and apoptosis, and can also play a role in cell proliferation or differentiation. MPs circulate in the bloodstream, can be detected in the peripheral blood, and may originate from different vascular cell types (eg, platelets, monocytes, endothelial cells, red blood cells, and granulocytes). SUMMARY: Cells of various types release small membrane vesicles called MP on their activation, as well as during the process of apoptosis. The properties and roles of MP generated in different contexts are diverse and are determined by their parent cell and the pathway of their generation, which affects their content. MP are involved in multiple cellular functions, including immunomodulation, inflammation, coagulation, and intercellular communication. MPs are able to deliver molecular signals in the form of lipids, proteins, nucleic acids, or functional trans-membrane proteins from the parent cell to distantly located targets. From a clinical point of view, MP may serve as biomarkers for disease status and may be found useful for developing novel therapeutic strategies.

43-50

Authors: Andri Hidayat, Andi Wijaya, Harun Alrasyid

BACKGROUND: Many studies have shown that obesity was closely related to insulin resistance via several pathways such as inflammation, oxidative stress, lipolysis, and endothelial dysfunction. This study was carried out to observe the correlation between inflammation (IL-6 and hsCRP), lipolysis process (ET-1), and endothelial dysfunction (ADMA) and insulin resistance (HOMA-IR) in centrally obese men. METHODS: This was a cross sectional study on 62 male subjects aged 30–60 years old with waist circumference (WC) >90 cm. IL-6, ET-1 and ADMA levels were measured using ELISA method, while hsCRP and insulin were measured using chemiluminescence method. All blood testings were conducted in Prodia Clinical Laboratory. RESULTS: The results showed that WC was significantly correlated with hsCRP (r=0.294, p=0.022 ), ET-1 (r=0.257, p=0.047 ) and ADMA (r=0.338, p=0.009). We also found a significant correlation between hsCRP with HOMA-IR (r=0.324, p=0.021), ADMA with HOMA-IR (r=0.280, p=0.045), and IL-6 with hsCRP (r=0.437, p=0.003). CONCLUSIONS: hsCRP and ADMA have significant correlation with HOMA-IR in centrally obese men. HOMA-IR significantly increases in subjects with ADMA above median and either IL-6 or hsCRP above median, as compared to those in the other groups. Inflammation and endothelial dysfunction are important causal pathways of insulin resistance state in centrally obese men.

57-63

Authors: Susana Gunawan, Anwar Santoso, Andi Wijaya

BACKGROUND: γ-GT is known as a primary enzyme that constituent extracellular glutathione catabolism. Various researches have proven that an increase in γ-GT concentration is related to oxidative stress and to components of metabolic syndrome. With individuals having metabolic syndrome as research subjects, our objective is to find the correlation of γ-GT, GPx and SAT. METHODS: This was a cross sectional study done on 36 subjects with metabolic syndrome who met the IDF 2005 criteria. Statistical analysis was carried out using SPSS for Windows ver.11.5 with a significance level of p<0.05. The correlation among biomarkers were assessed using Spearman’s Rho test. RESULTS: γ-GT concentration was correlated negatively with GPx concentration (r=-0.411, p=0.014), but correlated positively with hs-CRP concentration (r=0.385, p=0.022). The increase of γ-GT concentration had a potential risk 8.5 times higher than the increase of hs-CRP concentration, which proved to be more dominant in comparison to its risk to γ-GT/GPx or γ-GT/SAT ratios (p<0.05). CONCLUSIONS: An increase in γ-GT concentration characterizes signs of oxidative stress syndrome and of inflammation in individuals with metabolic syndrome.

92-112

Authors: Anna Meilina, Andi Wijaya

BACKGROUND: Plasma concentration of high density lipoprotein cholesterol (HDL-C) are strongly, consistenly, and independently inversely associated with risk of atheroschlerotic cardiovascular disease (CVD). However, the last decade has seen several observations that do not follow this simple script. CONTENT: A proteomic analysis of HDL has given us an intriguing glimpse into novel components of HDL. HDL isolated from normal humans contains several classes of proteins, including not only apolipoproteins, but also complement regulatory proteins, endopeptidase inhibitors, hemopexin, and acute phase response proteins. These observations raise the possibility of unsuspected roles for HDL. HDL delivery of complement proteins would implicate HDL in innate immunity. Serine proteinase inhibitors would enable HDL to modulate proteolysis of the vessel wall. HDL from patients with coronary artery disease was enriched in apoE, apoC-IV, apoA-IV, Paraoxonase (PON), and complement factor C3. Highlighted additional mechanisms through which HDL protects the vessel wall are: HDL improves vascular function, decreases vascular inflammation, detoxifies radicals, and limits thrombosis. SUMMARY: Both inter- and intra-organ desynchrony may be involved in the pathogenesis of cardiometabolic disease attributable to effects in brain and multiple metabolic tissues including heart, liver, fat, muscle, pancreas, and gut. Efforts to dissect the molecular mediators that coordinate circadian, metabolic, and cardiovascular systems may ultimately lead to both improved therapeutics and preventive interventions.

118-25

Authors: Anna Meiliana, Andi Wijaya, Suryani As'ad

BACKGROUND: The increased prevalence of obesity worldwide is correlated with increasing prevalence of metabolic syndrome. Studies of adipose tissue have been improved from an inert energy storage to a metabolic active endocrine organ. Adipokines secreted by this tissue play a role in maintaining metabolic homeostasis. The large mass of visceral fat tissue causing the imbalance of these adipokines leading to metabolic abnormality known as the metabolic syndrome (MetS). This study was performed to understand relationship of proinflammatory adipokines (resistin, TNF-α, RBP4 and visfatin) and anti-inflammatory adipokines (adiponectin and vaspin) in the development of MetS. METHODS: This was a cross-sectional study using 122 central obesity men with waist circumference >90 cm, age from 30–60 years old. Proinflammatory adipokines (resistin, TNF-α, RBP4 and visfatin) and anti-inflammatory adipokines (adiponectin and vaspin) was measured by ELISA method. RESULTS: The crosstab study showed that subjects who have >2 high proinflammatory adipokines (17.3%) has higher MetS prevalence (OR = 1.16; p = 0.72) compare to subjects with <2 high proinflammatory adipokines (14.8%), subjects with low anti-inflammatory adipokines profile (18.9%) has higher prevalence of MetS (OR=1.38; p=0.22) compare to subjects with high anti-inflammatory adipokines (13.7%) and the prevalence of MetS became 1.49 times higher (p=0.24) when we combine the high RBP4 and low adiponectin profile (21.1%) compare to subjects with low RBP4 and high adiponectin (14%). CONCLUSIONS: This study showed that each adipokine was not strong enough to induce MetS, so the interaction between proinflammatory and antiinflammatory adipokines were needed to induce a systemic metabolic abnormality. Thus, the adipokines equilibrium was important to prevent MetS especially in centrally obese subjects.

131-6

Authors: Waode Nurfina, Irawan Yusuf, Mansyur Arif

BACKGROUND: The low inflammatory state that accompanies the Metabolic Syndrome (MetS) associates with the overexpression of oxidative stress. Ferritin and Transferrin serum are often used to measure iron status and their concentrations are altered in several metabolic conditions. We hypothesized that concentration of Ferritin and Transferrin serum increase in Metabolic Syndrome (MetS) and correlate with the inflammation and oxidative stress. METHODS: We studied 65 male MetS patients, aged 43.26±7.16 years. Iron metabolism was measured by concentration of Ferritin and Transferrin serums, while inflammatory and oxidative stress by high sensitivity C-reactive Protein (hsCRP) and F2-Isoprostane. RESULTS: Concentration of Ferritin 315.70±188.63 ng/L and Transferrin 2.36±0.31 g/L increased along with increasing components of MetS. Concentration of Ferritin serum had a positive correlation with hsCRP (r=0.220) and F2-Isoprostane (r=0.023). CONCLUSION: Serum concentration of Ferritin increased in the MetS and correlates with hsCRP and F2-Isoprostane.

16-32

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Epidemiological data reveal parallel trends of decreasing sleep duration and increases in metabolic disorders such as obesity, diabetes and hypertension. There is growing evidence that these trends are mechanistically related. CONTENT: The circadian system orchestrates the temporal organization of many aspects of physiology, including metabolism, in synchrony with the 24 hours rotation of the Earth. The circadian system is a complex feedback network that involves interactions between the central nervous system and peripheral tissues. Circadian regulation is intimately linked to metabolic homeostasis and that dysregulation of circadian rhythms can contribute to disease. Conversely, metabolic signals also feed back into the circadian system, modulating circadian gene expression and behavior. SUMMARY: Both inter- and intraorgan desynchrony may be involved in the pathogenesis of cardiometabolic disease attributable to effects in brain and multiple metabolic tissues including heart, liver, fat, muscle, pancreas and gut. Efforts to dissect the molecular mediators that coordinate circadian, metabolic, and cardiovascular systems may ultimately lead to both improved therapeutics and preventive interventions.

40-5

Authors: Dwi Yuniati Daulay, Marsetio Donosepoetro, Sutomo Kasiman

BACKGROUND: Many studies have reported that diabetes mellitus correlates with vascular calcification event that increases progressively in uncontrolled diabetes. Osteoprotegerin (OPG) is known to act as a promoter in vascular calcification, contrary to Matrix Gla Protein (MGP), which is an inhibitor in vascular calcification. The aim of this study was to observe the progress of vascular calcification in uncontrolled diabetes patients by assessing biochemical markers OPG as promoter and MGP as inhibitor in vascular calcification. METHODS: This was an observational study with cross sectional design on adult male patients with type 2 diabetes mellitus, defined by DM Consensus Criteria Indonesia, 2006. RESULTS: The results of this study showed that there was a positive significant correlation between OPG and HbA1c (r=0.261, p=0.030), in contrast with MGP that showed no significant correlation with HbA1c. OPG also correlated significantly with Fasting Plasma Glucose (r=0.261, p=0.014). In uncontrolled diabetes group there was positive significant correlation between OPG and HbA1c (r=0.397, p=0.014). There was no significant difference found in the levels of OPG in controlled and uncontrolled diabetes groups (p=0.567), but OPG/MGP index showed higher difference (p=0.259). The OPG/MGP index also had positive significant correlation with HbA1c (r=0.285, p=0.018) and Fasting Plasma Glucose (r=0.313, p=0.009). CONCLUSIONS: This study suggested progress to vascular calcification in uncontrolled type 2 diabetes mellitus. The use of vascular calcification biomarkers are recommended to predict/detect vascular calcification event in type 2 diabetes mellitus patients.

53-8

Authors: Maria Diah Fibriani, Andi Wijaya, Burhanuddin Bahar

BACKGROUND: Obesity has a central role in the metabolic syndrome, which raises the risk for atherosclerotic cardiovascular disease (ASVCD). Apo B-48 and Apo B-100 are the necessary structural proteins required for the assembly and secretion of chylomicron and VLDL which have role in atherogenesis. The key initiating process in atherogenesis is the subendothelial retention of apolipoprotein B-containing lipoproteins. Oxidation of LDL is a hallmark of atherosclerosis development. The aim of this study was to asses the association between Apo B-48 and Apo B-100 with Oxidized-LDL as marker of atherosclerosis risk in central obesity. We hope that the result of this study can help to make a new strategy for the prevention and treatment of vascular disease. RESULTS: There were 68 patients aged 39.6±7.3 years, Apo B-48 concentration was 7.47±5.36 μg/mL, Apo B-100 was 117.26±25.74 mg/dL, and ox-LDL was 137.05±18.88 U/L. This study showed a significant correlation between Apo B-100 and ox-LDL (r=0.608, p<0.05) and correlation between Apo B-48 and ox-LDL (r=0.171, p<0.05). The levels of Apo B-100 were significantly different between obese with Mets and obese without Mets individuals (p<0.05). CONCLUSIONS: This study suggested that Apo B-100 concentration increase in obese in Mets as compared with obese without Mets. Apo B-48 and Apo B-100 were correlated with Oxidized LDL, but correlation between Apo B-100 and ox-LDL more significant that Apo B-48and ox-LDL.

26-44

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: There is probably no single way to age. Indeed, so far there is no single accepted explanation or mechanisms of aging (although more than 300 theories have been proposed). There is an overall decline in tissue regenerative potential with age, and the question arises as to whether this is due to the intrinsic aging of stem cells or rather to the impairment of stem cell function in the aged tissue environment. CONTENT: Recent data suggest that we age, in part, because our self-renewing stem cells grow old as a result of heritable intrinsic events, such as DNA damage, as well as extrinsic forces, such as changes in their supporting niches. Mechanisms that suppress the development of cancer, such as senescence and apoptosis, which rely on telomere shortening and the activities of p53 and p16INK4a may also induce an unwanted consequence: a decline in the replicative function of certain stem cells types with advancing age. This decrease regenerative capacity appears to pointing to the stem cell hypothesis of aging. SUMMARY: Recent evidence suggested that we grow old partly because of our stem cells grow old as a result of mechanisms that suppress the development of cancer over a lifetime. We believe that a further, more precise mechanistic understanding of this process will be required before this knowledge can be translated into human anti-aging therapies.

54-60

Authors: Ida Paulina Sormin, Widjaja Lukito, Andi Wijaya, Suryani As'ad

BACKGROUND: Biogenesis of HDL involves factors that regulate the synthesis, intravascular remodeling, and catabolism of HDL. Disturbance of these factors can lead to low concentration of HDL-C. Metabolic syndrome (MetS) is characterized by low concentration of high-density lipoprotein cholesterol (HDL-C). In MetS occur several pathological conditions including oxidative stress and impaired fibrinolysis, which contribute to the risk of atherosclerosis process. The correlation between oxidative stress and impaired fibrinolysis with HDL biogenesis dysfunction and its correlation with low concentration of HDL-C has not been well understood and therefore needs to be further investigated. METHODS: This study was an observational study with crosssectional design, involving 163 adult men, aged 25-60 years with metabolic syndrome. Concentration of apoA-1, prebeta-1 HDL, CETP, F2-isoprostan, PAI-1, and HDL-C were measured. The apo A1/HDL ratio indicated HDL maturation, whereas the CETP/HDL-C and CETP/TG ratios indicated HDL catabolism. RESULTS: The study showed that there were a positive correlation between PAI-1 with apoA1/HDL-C ratios (r=0.226, p=0.005) and a negative correlation with the CETP/TG ratios (r=-0.215, p=0.007), whereas F2-isoprostan did not have correlation with HDL biogenesis factors. CONCLUSIONS: We concluded that there was correlation between impaired fibrinolysis with decreased HDL maturation and there was increased HDL catabolism leading to low HDL-C concentration in men with metabolic syndrome.

66-72

Authors: Cynthia Retna Sartika, Andi Wijaya, Suryani As'ad

BACKGROUND: Central obesity is closely associated with chronic inflammation, characterized by abnormal cytokine production such as IL-1β, tumor necrosis factor-α (TNF-α) and tumor necrosis factor receptor-2 (TNFR-2). Central obesity and chronic inflammation form a complex link with insulin resistance, leading to the development of type-2 diabetes mellitus (T2DM) and cardiovascular disease (CVD). Knowing the coinciding occurrence of chronic inflammation and central obesity, this study aimed to examine pro-inflammatory cytokine profiles of Indonesian adult men with central obesity. METHODS: This cross-sectional study recruited 80 apparently healthy Indonesian adult men, aged 23-53 years with waist circumference of 64-125 cm. This study was done in Jakarta. Measurements included clinical parameters like systolic blood pressure (SBP), diastolic blood pressure (DBP), fasting blood glucose (FBG), aspartate amino transferase (AST), alanine amino transferase (ALT), creatinine, high sensitivity C-reactive protein (hsCRP), anthropometric parameters, namely weight, height and waist circumference; and pro-inflammatory cytokines TNF-α, soluble TNFR-2 (sTNFR-2) and IL-1β. RESULTS: Basic characteristics of the subjects showed linear increase in values of SBP, DBP and the serum concentrations of AST, ALT, FBG, GFR, hsCRP (p<0.005, respectively) with the degree of obesity. sTNFR-2 and IL-1 β positively correlated with hsCRP (r=0.277, p=0.013 and r= 0.257, p=0.022, respectively), WC (r=0.380, p=0.001 and r=0.400, p<0.001, respectively) and body mass index (BMI) (r=0.364, p=0.001 and r=0.399, p<0.001, respectively). Moreover, TNF-α did not show correlations with hsCRP, WC and BMI. CONCLUSION: There was a linear increase in the serum concentrations of sTNFR-2 and IL-1β in subjects with central obesity. Both pro-inflammatory markers, correlated with hsCRP, WC and BMI, but TNF-α did not. sTNFR-2 and IL-1β were, therefore, considered as valid biomarkers to indicate chronic inflammation in Indonesian adult men with central obesity.

33-56

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: The prevalence of obesity is increasing in both developed and developing countries along with associated diseases such as type 2 diabetes and coronary heart disease. The recent discovery of a number of gut hormones that play a role in appetite regulation and are released or suppressed in response to a meal may offer new targets for the treatments of obesity. CONTENT: In addition to the obvious role of the gut in the digestion and absorption of nutrient, the intestine and associated visceral organs, including the pancreas, liver, and visceral adipose depots, have important sensing and signaling roles in the regulation of energy homeostatis. Signals reflecting energy stores, recent nutritional state, and other parameters are integrated in the central nervous system, particularly in the hypotalamus, to coordinate energy intake and expenditure. SUMMARY: Our understanding of the role of the gut in energy balance and insights into gut-derived signals will stimulate previously unexplored therapeutics for obesity and other disorders of energy balance.

65-70

Authors: Johnson Wijaya, Marsetio Donosepoetro, Syakib Bakri

BACKGROUND: Vascular remodeling was an adaptive process of the vascular wall that occured in response to long-term changes in hemodynamic conditions that contribute to the changes of the vascular structure and the pathophysiology of vascular disease. On the other hand, Endothelial Progenitor Cells (EPC) derived from bone marrow had the capacity to migrate to the peripheral circulation and to differentiate into mature endothelial cells. Therefore, EPC could contribute in the endothelial repairing after endothelial injury. METHODS: This study was a cross sectional design. Analysis was done among 30 subjects with normoalbuminuria normotension, 55 subjects with normoalbuminuria hypertension and 30 subjects with with microalbuminuria hypertension. TGF-β1, MMP-9 and VEGFR-2 testing were performed by ELISA method. All statistical calculations were performed using the SPSS 11.5 statistical software package. We used the Independent sample T test, Mann-Whitney, One Way Annova and Kruskal Wallis to establish the difference among various biochemical measures. RESULTS: TGF-β1 concentration was increased from normoalbuminuria normotension to normoalbuminuria hypertension and to microalbuminuria hypertension (27.63178±12.97246 vs. 38.61193±17.09546 vs. 38.73939±12.63911 ng/mL). TGF-β1 concentration was higher significantly in normotension as compared to hypertension (27.63178±12.97246 vs. 38.65692±15.58950, p<0.001) and to microalbuminuria hypertension (38.73939±12.63911, p<0.001). MMP-9 concentration was increased in normotension to normoalbuminuria hypertension but was decreased in microalbuminuria hypertension (438.1967±156.4268 vs. 564.5873±291.2876 vs. 418.6900±188.3801 ng/mL). MMP-9 concentration was higher significantly in normoalbuminuria hypertension as compared to microalbuminuria hypertension (p=0.028). VEGFR-2 concentration was decreased from normotension to normoalbuminuria hypertension to microalbuminuria hypertension (9.90552±1.85185 vs. 9.39561±1.75413 vs. 9.00506±1.47173 ng/mL). VEGFR-2 concentration was higher significantly in normotension as compared to microalbuminuria hypertension (p=0.042). CONCLUSIONS: The increasing concentration level of TGF-β1 and decreasing concentration level of MMP-9 in microalbuminuria hypertension showed that the remodeling process was getting increased. The decreasing concentration level of VEGFR-2 in microalbuminuria hypertension showed that the repairing process was getting decreased.

76-80

Authors: Yani Lina, Gatot Susilo Lawrence, Andi Wijaya, Suryani As'ad

BACKGROUND: Fibroblast growth factor-21 (FGF21) is known as an important endocrine and paracrine regulator of metabolic homeostasis. Recent studies have shown that FGF21 attenuates lipolysis in human adipocytes, which is suggested as a FGF21's mechanism as anti-hyperlipidemia, anti-hyperglycemia and anti-obesity. The aim of this study was to measure the correlation between FGF21, FFA, hsCRP and HOMA-IR among Indonesian obese non diabetic males. METHOD: The study was observational with cross sectional design. The analysis was done in 137 subjects aged 30-60 years with non diabetic abdominal obesity. We measured the biochemical markers FGF21, FFA, hsCRP, fasting insulin and fasting glucose. We also measured weight, height, waist circumrefence (WC), creatinine, serum glutamin oxaloacetic transaminase (SGOT), and serum glutamic pyruvic transaminase (SGPT), systolic blood pressure (SBP) and diastolic blood pressure (DBP). Correlation between markers was measured using Pearson and Spearman's analysis. RESULT: There were significant positive correlations between FGF21-HOMA-IR (r=0.314, p=0.000); FGF21-WC (r=0.173, p=0.043); FFA=hsCRP (r=0.270, p=0.001); and WC-HOMA-IR (r=0.279, p=0.001). There was significant negative correlation between FGF21-FFA (r=-0.038, p=0.657) and FGF21-hsCRP (r=-0.061, p=0.482). CONCLUSION: In this study we found that although there was no significant correlation, FGF21 might act as an anti-lipolytic and anti-inflammation agent among Indonesian obese non-diabetic males. Our findings agree with results of previous studies that the positive correlation between FGF21-WC and FGF21-HOMA-IR moght occur as a compensatory mechanism or resistance to FGF21 in obesity.

88-96

Authors: Siti Nurul Mubarokah, I Dewa Agung Susilawati, Sumarno Sumarno, I Ketut Gedhe Muliartha, Djanggan Sargowo

BACKGROUND: Periodontitis is caused mostly by Porphyromonas gingivalis (P.gingivalis) and it is related to acute coronary syndrome. P.gingivalis readily invades blood circulation and potentially induces collagenolytic activity of inflammatory cells that results in collagen vascular degradation leading to atherosclerotic plague rupture (APR). APR is responsible for the occurence of fatal cardiovascular events such as acute myocardial infraction (AMI). AIMS: To show that P.gingivalis potentially induces fragmentation of the type IV vascular collagen due to macrophage-activated MMP-9. MATERIAL AND METHODS: The ability of P.gingivalis to induce the type IV collagen fragmentation, shown by digesting type IV collagen with the supernatant of monocyte-derived macrophage activated by exposure to P.gingivalis suspension for 18 hours, 37oC, 5%CO2. The type IV collagen fragments were analyzed by SDS-PAGE and confirmed by Western-blotting. Antibody of type IV collagen produced and confirmed by dot-blotting prior to its being used as primary antibody of Western-blotting. The existence of MMP-9 was detected by Dot-blot and Western-blot technique, while the MMP-9 activity was assessed by SDS-PAGE and zymograms. RESULTS: Our data showed that P.gingivalis induced macrophage to produce MMP-9 as one of collagenolytic components, and interaction with P.gingivalis proteases enhanced the proteolytic activity and resulted in degradation of type IV collagen with molecular weight of 88 kDa into two smaller fragments with molecular weight of 80 kDa and 60 kDa. CONCLUSION: P.gingivalis induced macrophage to activate its MMP-9 that led to fragmentation of vascular type IV collagen in the pathogenesis of atherosclerotic plaque rupture.