The Indonesian Biomedical Journal

123-36

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Prostate cancer (PCa) was the second most common type of cancer and the fifth leading cause of cancer-related death in men. The great challenge for physicians is being able to accurately predict PCa prognosis and treatment response in order to reduce PCa-speciic mortality while avoiding overtreatment by identifying of when to intervene, and in which patients. CONTENT: Currently, PCa prognosis and treatment decision of PCa involved digital rectal examination, Prostate-Speciic Antigens (PSA), and subsequent biopsies for histopathological staging, known as Gleason score. However, each procedure has its shortcomings. Efforts to find a better clinically meaningful and non-invasive biomarkers still developed involving proteins, circulating tumor cells, nucleic acids, and the ‘omics' approaches. SUMMARY: Biomarkers for PCa will most likely be an assay employing multiple biomarkers in combination using protein and gene microarrays, containing markers that are differentially expressed in PCa.

157-62

Authors: Ferry Sandra, Roya Khosravi-Far

BACKGROUND: Our previous report showed that phosphorylated-survivin at Ser81 induces survivin back loop to activate protein kinase A (PKA) in the cytoprotection mechanism. Activated PKA could possibly induce the cytoprotection via Phosphatydilinositol 3-kinase (PI3K). Therefore our current study was conducted to investigate the possibility of survivin-PKA-PI3K signaling pathway. METHODS: Viral productions by BOSC23 cells of Survivin, Antisense Survivin (Survivin-AS) and Ser81Ala mutant (Survivin-S81A) in pMSCV-IRES-GFP vector with cytomegalovirus (CMV) promoter were conducted. L929 cells were pretreated with/without PKI 6-22 amide and infected with viral particle of Survivin, Survivin-AS, Survivin-S81A or vector only. Cells were harvested, lysed and immunoprecipitated with anti-PI3K (p85) antibody and immunoblotted to detect PI3K (p85) and phospho-(Tyr) p85 PI3K. To confirm PI3K activation, PI3K Activity Assay was conducted by using phosphoinositide fraction containing PtdIns(4,5)P2 and [32P]ATP. RESULTS: Immunoblot and PI3K activity results showed similar results. Upon infection of virus with survivin, a markedly increased level of tyrosine phosphorylation of p85 PI3K or PI3K activity in L929 cells was seen. Low levels of tyrosine phosphorylation of p85 PI3K or PI3K activity were observed for Survivin-AS and Survivin-S81A-viral-infected L929 cells. With higher concentrations of Survivin-viral-infection, levels of tyrosine phosphorylation of p85 PI3K or PI3K activity in L929 cells were gradually increased. However, when L929 cells were pretreated with PKI 6-22 amide, prior to Survivin-viral-infection, level of tyrosine phosphorylation level of p85 PI3K or PI3K activity was detected much lower. CONCLUSION: Our result suggest that Ser81 Survivin play role in inducing PI3K activation and the Survivin-PI3K signaling pathway was PKA-dependent.

167-74

Authors: Benyamin Lukito, Ivet Suriapranata, Pendrianto Pendrianto, Ali Sulaiman, Irawan Yusuf, FX Budhianto Suhadi, George Mathew, Ferry Sandra

BACKGROUND: Alpha-fetoprotein (AFP) has been used for hepatocellular carcinoma (HCC) diagnosis and screening, however, AFP has poor specificity. The extensive hypervascularity associated with HCC could be driven in part by the pro-angiogenic factor known as vascular endothelial growth factor (VEGF). Furthermore, invasiveness of certain HCC lesions has recently been linked to high levels of VEGF. Therefore, circulating VEGF levels of patients with liver cirrhosis (LC) and HCC were investigated and analysed. METHODS: An analytical cross sectional study was designed. Diagnosis of HCC and LC was performed using clinical criteria and findings obtained from B-mode ultrasonography (USG), computed tomography (CT) angiography, or magnetic resonance imaging (MRI). Blood were collected intravenously from all subjects. Obtained serum and plasma were stored in -80°C for following analyses: hepatitis B surface antigen (HBSAg), hepatitis C virus (HCV), alanine aminotransferase (ALT), total bilirubin, albumin, VEGF and AFP. RESULTS: Levels of VEGF and AFP were significantly higher in HCC group compared with LC group with p = 3.05 x 10-6 and p = 8.74 x 10-5, respectively. There was a significant positive correlation (p=0.029, r=0.309) between VEGF level and tumor size in HCC group. The area under curve (AUC) for VEGF level in HCC and LC groups was 0.771. In the level of median 435.6 pg/mL VEGF, the sensitivity was 50% and specificity was 86%. In the level of 199.99 pg/mL VEGF the sensitivity was 74% and specificity was 76%. CONCLUSION: The present findings suggested that VEGF level could be a useful marker for the presence of HCC in patients with LC.

79-90

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Over the past 50 years, scientists have recognized that not all adipose tissue is alike, and that health risk is associated with the location as well as the amount of body fat. Different depots are sufficiently distinct with respect to fatty-acid storage and release as to probably play unique roles in human physiology. Whether fat redistribution causes metabolic disease or whether it is a marker of underlying processes that are primarily responsible is an open question. CONTENT: The limited expandability of the subcutaneous adipose tissue leads to inappropriate adipose cell expansion (hypertrophic obesity) with local inflammation and a dysregulated and insulin-resistant adipose tissue. The inability to store excess fat in the subcutaneous adipose tissue is a likely key mechanism for promoting ectopic fat accumulation in tissues and areas where fat can be stored, including the intra-abdominal and visceral areas, in the liver, epi/pericardial area, around vessels, in the myocardium, and in the skeletal muscles. Many studies have implicated ectopic fat accumulation and the associated lipotoxicity as the major determinant of the metabolic complications of obesity driving systemic insulin resistance, inflammation, hepatic glucose production, and dyslipidemia. SUMMARY: In summary, hypertrophic obesity is due to an impaired ability to recruit and differentiate available adipose precursor cells in the subcutaneous adipose tissue. Thus, the subcutaneous adipose tissue may be particular in its limited ability in certain individuals to undergo adipogenesis during weight increase. Inability to promote subcutaneous adipogenesis under periods of affluence would favor lipid overlow and ectopic fat accumulation with negative metabolic consequences.

97-100

Authors: Yesaya Yunus, Julius July, Andi Asadul Islam, Eka Julianta Wahjoepramono, Irawan Yusuf

BACKGROUND: Inflammation following aneurysmal subarachnoid hemorrhage (SAH) is associated with patient outcome. This study aimed to analyze the dynamics pattern of the pro-inflammatory interleukin (IL)-1β and anti-inflammatory cytokine IL-10 in the cerebrospinal fluid (CSF) following aneurysmal SAH. METHODS: This is a prospective observational study. The CSF was collected prior to surgery, and on the 2nd, 4th and 6th days after surgery. The CSF was then analyzed for IL-1β and -10 using enzyme-linked immunosorbent assay. Delayed Ischemic Neurological Deficits (DIND) was determined based on new neurological deficits within the first week after surgery. Data was analysed with T test or Wilcoxon Rank-Sum test to evaluate the pattern of biomarkers between DIND and non-DIND groups. RESULTS: There were 33 patients enrolled in this study, 16 patients (48.48%) who experienced DIND and 17 patients (51.52%) were non-DIND. There were significant increase dynamic levels of IL-10 and -1β in DIND patients (p<0.05). CONCLUSION: Significant increase levels of IL-10 and -1β in CSF after aneurysmal SAH are associated with DIND.

107-12

Authors: Linda Lison, Irawan Yusuf, Bambang Sutrisna, Peter Kabo, Teguh Santoso, Ferry Sandra

BACKGROUND: Syntax score is a lesion-based angiographic scoring system, originally devised to grade the anatomic complexity of coronary artery disease (CAD) and thereby facilitate consensus in the study of a diagnostic angiogram between surgeons and interventional cardiologists. Suitable biology marker to predict the severity of CAD and to have a treatment decision for revascularization by percutaneous coronary intervention (PCI) or bypass surgery, is still unknown. Which biology marker can be a predictor for higher Syntax score remains unknown as well. METHODS: Severity of 105 patients who underwent coronary angiography, were evaluated with Syntax score. Laboratory tests were conducted for routine blood test, serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), high-sensitivity C-reactive protein (hs-CRP), total cholesterol, low-density lipoprotein (LDL) cholesterol, high-density lipoprotein (HDL) cholesterol, triglycerides, kidney function, creatinine and hemoglobin A1c (HbA1c). RESULTS: Ureum had significant correlation with Syntax score, p=0.016 and 95% confidence interval (CI)=0.05-0.55. Result of logistic regression analysis showed that HDL and haematocrit had significant correlation with Syntax score, with cut off point 34 (p=0.02, p=0.0033, respectively). Based on multi-variate analysis, Syntax score 34 = 16 (HDL ≤40 + Ureum ≥23) + 10 (Neutrophil/Lymphocyte ≥3.789) +10 (HbA1c ≥7 ) + 13 ( monocyte ≥9%). CONCLUSION: HDL cholesterol, ureum, neutrophil-lymphocyte ratio, HbA1c and monocyte altogether can predict the higher Syntax score.

17-32

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: The strong inverse association of plasma levels of high-density lipoprotein (HDL) cholesterol with coronary heart disease (CHD) found in human epidemiological studies led to the development of the ‘HDL cholesterol hypothesis’, which posits that intervention to raise HDL cholesterol will result in reduced risk of CHD. A number of recent developments have brought the potential protective role of HDL into question. Several clinical trials of agents that substantially raise HDL-C have been demonstrated to not reduce CHD event rates. CONTENT: For decades, HDL and HDL-cholesterol (HDL-C) levels were viewed as synonymous, and modulation of HDL-C levels by drug therapy held great promise for the prevention and treatment of cardiovascular disease. Nevertheless, recent failures of drugs that raise HDL-C to reduce cardiovascular risk and the now greater understanding of the complexity of HDL composition and biology have prompted researchers in the field to redefine HDL. As such, the focus of HDL has now started to shift away from a cholesterol-centric view toward HDL particle number, subclasses, and other alternative metrics of HDL. Many of the recently discovered functions of HDL are, in fact, not strictly conferred by its ability to promote cholesterol flux but by the other molecules it transports, including a diverse set of proteins, small RNAs, hormones, carotenoids, vitamins, and bioactive lipids. Based on HDL’s ability to interact with almost all cells and deliver fat-soluble cargo, HDL has the remarkable capacity to affect a wide variety of endocrine-like systems. SUMMARY: There is a significant need to redefine HDL and its benefit. HDL transports a diverse set of functional proteins, including many binding proteins. HDL transports and deliver vitamins, carotenoids, and other small molecules. Moreover, HDL transports hormones, steroids and bile acids, and can modulate multiple endocrine pathways. HDLs also transport and deliver microRNAs to recipient cells and control gene expression. Likewise, HDLs carry bioactive lipids and can activate signaling cascades and receptors that control endothelial apoptosis, migration, survival and activation. Many of HDL’s alternative noncholesterol cargo likely confer many of HDL’s alternative functions.

45-50

Authors: Rusli Muljadi, Bachtiar Murtala, Peter Kabo, FX Budhianto Suhadi

BACKGROUND: Cardiovascular disease is one of the atherosclerosis etiologies that can lead to death. Diabetes mellitus increases the risk of atherosclerosis. Screening tool is very beneficial for detecting atherosclerotic plaque, especially in subclinical atherosclerotic cases. Carotid intima-media thickness (CIMT) and coronary artery calcification score (CACS) are two kinds of tools that are widely used, and each of these tools has its own superiority. This study was aimed to investigate the sensitivity and specificity of both of these tools as screening tools. METHODS: The study was conducted with a cross sectional design involving 43 diabetic and 68 non-diabetic male subjects aged above 45 years old. All subjects fulfilled inclusion criteria. Carotid artery ultrasonography and CACS measurement were performed. RESULTS: Fischer exact test was used to show a significant correlation between CIMT and CACS (p<0.05). Diagnostic test was used to assess the sensitivity of CIMT toward CACS in above 75 percentile. The left common carotid artery (LCCA) showed the highest sensitivity either in diabetic (76.4%) or non-diabetic male subjects (90%). CONCLUSION: CIMT has the same sensitivity with CACS. CIMT can be used as the preferred screening tool for high risk patients and as a substitution tool to CACS for low risk patients in subclinical atherosclerosis detection.

57-62

Authors: Ferry Sandra, Janti Sudiono, Elina Ardiani Sidharta, Elisabeth Pricilia Sunata, Dea Jane Sungkono, Yanni Dirgantara, Angliana Chouw

BACKGROUND: Secreted factors contained in conditioned media (CM) of human umbilical cord blood mesenchymal stem cells (hUCB-MSCs) known as secretome, was suspected to have important roles in regulating cells. This study was conducted to investigate the role of CM-hUCB-MSCs-derived secretome in apoptosis and growth of HeLa cells. METHODS: HeLa cells were treated with secretome in various concentrations (0, 0.2, 2 and 20%) for 24 and 48 hours. Trypan blue exclusion assay was performed to detect cell viability. Meanwhile sub-G1 apoptotic assay was performed to detect apoptotic cells. The transition of mitochondrial transmembrane potential (TMP), which occurs in the apoptotic process, was analyzed by mitochondrial membrane potential (ΔΨM) assay. Both sub-G1 and ΔΨM assays were performed using FACSCanto flow cytometer. Statistical analyses were conducted using IBM SPSS Statistics to detect significance level at p<0.05. RESULTS: Secretome significantly induced cell death starting at concentration of 0.2% within a 24-hour period (p<0.05). Secretome significantly induced cell death in concentration and time dependent manner (p<0.05). The cell death was then confirmed as apoptosis through sub-G1 analysis. Due to the underlying apoptotic mechanism, we found distinct decrease of TMP, indicating an increase in mitochondrial membrane permeability of HeLa cells. In addition, we found that HeLa cell growth was inhibited partially by secretome. CONCLUSION: Taken together, we conclude that CMhUCB-MSCs-derived secretome significantly induced apoptosis of HeLa cells in a concentration and time dependent manner through mitochondrial apoptotic pathway. The secretome might also play important role in inhibiting HeLa cell growth.

151-60

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Blood vessels are a source of stem and progenitor cells, which likely contribute to a variety of vascular processes and diseases. Emerging concepts in this field could influence therapeutic approaches to diseases of blood vessels such as atherosclerosis. CONTENT: Vascular Stem Cells (VSCs) field is only beginning to emerge, and thus, many issues regarding VSCs’s identity and function remain poorly understood. In fact, even after decades of intensive research, Mesenchymal Stem Cells (MSC), which is suggested to be VSCs, is still having many outstanding issues of its own. And, on top of this, likewise decades-long intensive pericyte research has not been able resolve the identity issue. While favors Adventitial Progenitor Cells (APCs) over pericytes as the likely VSC candidate, it should be pointed out that currently the opposite view (i.e., pericytes as VSCs) is more prevalent, and many excellent reviews, including a recent one, have discussed this issue extensively. SUMMARY: It has been postulated that, within the vasculature, APCs could differentiate into pericytes (CD34- CD31- CD140b+ SMA-), endothelial cells (CD34+ CD31+ CD140b- SMA-), and smooth muscle cells (SMCs) (CD34- CD31- CD140b- SMA+); and during tissue expansion or repair, APCs could also differentiate into tissue-specific cell types (e.g., muscle and fat) Thus, in vitro, APCs fulfill all criteria for being VSCs. Meanwhile, in vivo evidence is still limited and will require further investigation.

169-72

Authors: Maria Evi Novianti, Syakib Bakri, Mansyur Arif, Ferry Sandra

BACKGROUND: Not only hypertension, prehypertension has been reported to be linked with increased cardiovascular morbidity and mortality risks as well. Prehypertension has three-fold hypertension and two-fold cardiovascular risks. Pathomechanism that links hypertension with cardiovascular is related with endothelial dysfunction and insulin resistance. Endothelial dysfunction occurs when nitric oxide (NO) biological function was impaired, whereas shown by asymmetric dimethylarginine (ADMA). Subjects with prehypertension had higher insulin resistance events than normotension, whereas shown by homeostatic model assessment-estimated insulin resistance (HOMA-IR). This research was conducted to investigate the correlation of HOMA-IR with ADMA in prehyper- and normo-tension. METHODS: A cross-sectional comparative research was designed. Subjects were recruited and divided into prehyper- and normo-tensive groups. ADMA was measured using ELISA method, while HOMA-IR was calculated by the ratio of fasting insulin and glucose. Spearman 1-tail and Mann Whitney statistical analyses were performed. RESULTS: Comparing to normotensive group, elevated levels of HOMA-IR and ADMA in prehypertensive group were shown, but not significant. In prehypertensive group, we found significant correlation between HOMA-IR and ADMA. CONCLUSION: Insulin resistance and endothelial dysfunction was elevated in prehyper-compared to normotension.

179-84

Authors: John Butar Butar, Zola Wijayanti, Beatrix Tjahyana, Veli Sunggono, Hori Hariyanto

BACKGROUND: This study was carried out to investigate the association of Cross Linked C-Telopeptide Type I & II Collagen (CTX-I and II) and hyaluronic acid (HA) with knee osteoarthritis (OA) severity. METHODS: Sixty menopause women with primary knee OA were enrolled in this study during their visits to the Outpatient Department. Patients with knee pain during weight bearing, active or passive range of motion, or tenderness with Kellgren-Lawrence (KL) grade of more than I were included. Patients with injury, inflammatory and metabolic diseases were excluded. Patients were put in a 10-hour fasting prior to withdrawal of morning blood samples for examinations of HA, CTX-I, interleukin 1 beta (IL-1β), and high sensitivity C reactive protein (hs-CRP) level. Second void morning urine specimens were taken for CTXII assessment. HA, CTX-I and II levels were measured by enzyme-linked immunosorbent assay. RESULTS: Sixty menopausal female patients were included in this study, 35 with KL grade II, 17 grade III, and 8 grade IV. Means of CTX-II were significantly different between subjects KL grade IV and III (p=0.021). Correlation of KL grade was significant with CTX-II (p=0.001, r=0.412) and HA (p=0.0411, r=0.269). KL grades were not significantly associated with CTX-I (p=0.8364, r=-0.0272); IL-1β (p=0.5773, r=0.0853) and hs-CRP (p=0.2625, r=0.1470). CONCLUSION: CTX-II and HA were associated with severity of knee OA, suggesting that CTX-II and HA can be used as marker for knee OA severity.

81-90

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Obesity is now recognized as the main cause of the worldwide epidemic of type 2 diabetes. Obesity-associated chronic inflammation is a contributing key factor for type 2 diabetes and cardiovascular disease. Numbers of studies have clearly demonstrated that the immune system and metabolism are highly integrated. CONTENT: Macrophages are an essential component of innate immunity and play a central role in inflammation and host defense. Moreover, these cells have homeostatic functions beyond defense, including tissue remodeling in ontogenesis and orchestration of metabolic functions. Diversity and plasticity are hallmarks of cells of the monocyte-macrophage lineage. In response to interferons (IFNs), toll-like receptor (TLR), or interleukin (IL)-4/IL-13 signals, macrophages undergo M1 (classical) or M2 (alternative) activation. Progress has now been made in defining the signaling pathways, transcriptional networks, and epigenetic mechanisms underlying M1, M2 or M2-like polarized activation. SUMMARY: In response to various signals, macrophages may undergo classical M1 activation (stimulated by TLR ligands and IFN-γ) or alternative M2 activation (stimulated by IL-4/IL-13); these states mirror the T helper (Th)1–Th2 polarization of T cells. Pathology is frequently associated with dynamic changes in macrophage activation, with classically activated M1 cells implicate in initiating and sustaining inflammation, meanwhile M2 or M2-like activated cells associated with resolution or smoldering chronic inflammation. Identification of the mechanisms and molecules that are associated with macrophage plasticity and polarized activation provides a basis for macrophage centered diagnostic and therapeutic strategies.

101-6

Authors: Ritawaty Ritawaty, Indriyanti Rafi Sukmawati, Ilhamjaya Patellongi, Ferry Sandra

ACKGROUND: Gamma glutamyltransferase (GGT) was reported recently to be associated with inflammation, oxidative stress and increased amino acid. However, role of GGT in insulin resistance pathomechanism is not exactly known. Therefore correlation of GGT with inflammation, oxidative stress and elevated amino acid, in men with central obesity need to be confirmed. METHODS: A cross-sectional study was designed. Men with central obesity were recruited and selected. Anthropometric parameters, creatinine, hs-CRP, fasting glucose, fasting insulin, glutathione peroxidase (GPx) activity, GGT, plasma total cysteine (tCys) and fatty liver were measured. Subjects were then divided in 4 groups based on waist circumference (WC) and fatty liver: Group I: WC ≤100 cm, without fatty liver; Group II: WC ≤100 cm, with fatty liver; Group III: WC >100 cm, without fatty liver; Group IV: WC >100 cm, with fatty liver. All biochemical characteristics in each group were then statistically analyzed. RESULTS: Seventy-two men with central obesity were selected. Numbers of subjects in each group were: Group I: n=33; Group II: n=5; Group III: n=17; Group IV: n=17. We found significant difference of HOMA-IR between Group I and IV, significant correlation between GGT and HOMAIR, and significant negative correlation between tCys with HOMA-IR in Group IV. CONCLUSION: GGT was significantly correlated with HOMA-IR in men with WC >100 cm and fatty liver. Further investigation with more subjects is necessary to determine clear GGT cut-off to distinguish subjects with fatty liver and insulin resistance.

115-20

Authors: Adriana Todingrante, Mansyur Arif, Uleng Bahrun, Ferry Sandra

BACKGROUND: There is a prevalence increase of overweight and obesity in Indonesia. Central obesity can lead a variety of chronic diseases through the inflammatory process. There are some markers for low-grade chronic inflammatory, such as cathepsin S, high sensitivity C-reactive protein (hs-CRP), interleukin-1- beta (IL-1β). To our current interest that central obesity can lead to various chronic diseases through the inflammatory process, we conducted a study to investigate correlation of Cathepsin S, hs-CRP, IL-1β in men with central obesity. METHODS: A cross-sectional study was conducted. Seventy-eight selected subjects were examined to collect anthropometric data and prepared for sample collection. Collected samples were processed for the following biochemical analyses: fasting glucose, high density lipoprotein (HDL)-cholesterol, triglyceride, serum glutamic oxaloacetic transaminase (SGOT), serum glutamate pyruvate transaminase (SGPT), cathepsin S, hs-CRP, and IL-1β. Data distribution and variable correlation were then statistically analyzed. RESULTS: There were significant correlations between waist circumference (WC) and cathepsin S (p=0.030; r=0.214), hs-CRP and cathepsin S (p=0.007; r=0.276), triglyceride and IL-1β (p=0.019; r=-0.235), WC and systolic blood pressure (SBP) (p=0.003; r=-0.312), WC and fasting glucose (p=0.000; r=0.380), WC and body mass index (BMI) (p=0.000; r=0.708). CONCLUSION: Our study showed that cathepsin S was correlated with central obesity, suggesting that cathepsin S could be a potential inflammatory marker in central obesity in the future.

129-36

Authors: Wiwik Rositawati, Syakib Bakri, Gatot Susilo Lawrence, Andi Wijaya

BACKGROUND: Vascular calcification may be an important risk factor of cardiovascular disease in Chronic Kidney Disease (CKD). The pathobiology of vascular calcification in CKD is complex and involves some factors including inflammation, oxidative stress and balancing of calcification regulators. The aim of the study was to investigate the interaction between 8-isoprostane with calcification regulators such as matrix gla protein (MGP) and bone morphogenetic protein (BMP)-2 with vascular calciication in CKD. METHODS: A cross-sectional study was performed on 63 subjects undergoing haemodialysis maintenance for more than 3 months. Abdominal aortic calcification (AAC) was measured using a lateral abdominal X-ray for calcification in abdominal aorta, which is related to severity of calcific deposits at lumbar vertebral segment (L)1-L4. Serum levels of 8-isoprostane, MGP and BMP-2 were measured by enzyme-linked immunosorbent assay method. RESULTS: Results showed that 8-isoprostane levels were correlated with BMP-2 (r=0.266, p=0.018) and MGP levels (r=0.410, p≤0.001). MGP/BMP-2 levels ratio was correlated with AAC score (r=0.279, p=0.013). Subjects were then stratified into 3 groups based on AAC score: 0, 1-6 and ≥7. The highest mean of MGP levels was in AAC score 1-6 group, and the highest mean of 8-isoprostane levels was in AAC score ≥7. In the group of AAC score 0, 8-isoprostane levels were correlated with MGP levels (r=0.499, p=0.001) and MGP/BMP-2 levels ratio (r=0.291, p=0.034). In AAC score 1-6 group, 8-isoprostane levels were correlated with BMP-2 (r=0.661, p=0.005) and MGP levels (r=0.569, p=0.017). In AAC score ≥7 group, MGP levels were positively correlated with AAC score (r=0.608, p=0.041). With multivariate logistic regression analyses, we identified that increased MGP/BMP-2 levels ratio (OR=12.436; 95% CI=1.595-96.971) was an independent predictor in aortic calcification event. CONCLUSION: We concluded that regulators of calcification including calcification inhibitor and promoter related to oxidative stress, were associated with vascular calcification in CKD. MGP levels were increased in the early of calcification and MGP/BMP-2 levels ratio was a strong predictor for vascular calcification in CKD.