The Indonesian Biomedical Journal

168-84

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: Increasing prevalence of obesity gives rise to many problems associated with multiple morbidities, such as diabetes, hypertension, heart disease, sleep apnea and cancer. The mechanism of obesity is very complex, thus its link to various disease is poorly understood. This review highlights important concepts in our understanding of the pathogenesis of obesity and related complications. CONTENT: Many studies have tried to explore the exciting and puzzling links between metabolic homeostasis and inflammatory responses. A form of subclinical, low-grade systemic inflammation is known to be associated with both obesity and chronic disease. This, later called as "metaflammation", refers to metabolically triggered inflammation. The nutrient-sensing pathway and the immune response coordination are facilitated by these molecular sites in order to maintain homeostasis under diverse metabolic and immune conditions. Recent studies have found that the NLRP3 inflammasome during metabolic stress forms a tie linking TXNIP, oxidative stress, and IL-1β production. This provides new opportunities for research and therapy for the disease often described as the next global pandemic: type 2 diabetes mellitus (T2DM). SUMMARY: The crucial role of metaflammation in many complications of obesity shown by the unexpected overlap between inflammatory and metabolic sensors and their downstream tissue responses. Then great interest arose to explore the pathways that integrate nutrient and pathogen sensing, give more understanding in the mechanisms of insulin resistance type 2 diabetes, and other chronic metabolic pathologies. A family of intracellular sensors called NLR family is a critical component of the innate immune system. They can form multiprotein complexes, called inflammasome which is capable of responding to a wide range of stimuli including both microbial and self molecules by activating the cysteine protease caspase-1, leading to processing and secretion of the proinflammatory cytokines IL-1β and IL-18, which play crucial roles in host defense. Inflammasome dysregulation has been linked to some autoinflammatory and metabolic diseases. These provide opportunities to continue to improve our understanding of the nature of metaflammation in the hope of modifying it to prevent and treat diseasese.

191-9

Authors: Ika Kustiyah Oktaviyanti, Djanggan Sargowo, Mohammad Aris Widodo, Karyono Mintaroem

BACKGROUND: NSAIDs can cause gastric ulcer or may delay the healing of it. Upon exposure to indomethacin, gastric ulcer can occur due to oxidants. Mangosteen rind contains xanthone, which is a natural antioxidant. Administration of this antioxidant may increase angiogenesis that can accelerate healing of gastric ulcer. METHODS: This study used an experimental method with randomized post test control only design using Wistar rats. The rats were put on fasting for 24 hours, then a single dose of 30mg/kg body weight (BW) Indomethacine was given. The rats were divided into control group and treatment group. The treatment group was further divided into two subgroups: one group was given a daily 200 mg/kg BW mangosteen pericap extract, and the other group was given 35 mg/kg BW Xanthone. Both the control group and treatment group were decapitated on the 3rd day, 6th day and 12th day, respectively. After decapitation, the stomach of each rat was taken and divided into two portions, one portion was used for NO examination by ELISA, and the other portion for hispathological examination and immunohistochemical analysis for assessing CD 31 and VEGF expressions. RESULTS: Administration of mangoosteen pericap and xanthone could accelerate healing of gastric ulcers as compared with the control, as shown by the decrease in the severity level of the ulcers. Mangoosteen pericap and xanthone could also increase NO, VEGF expression, and CD-31 as compared with the control, especially on the 3rd day of treatment. Explanation of this finding might be that the antioxidants contained in the mangoosteen pericap or in xanthone could bind with radical superoxide and accelerate release of free NO. The increase of NO caused increase of VEGF and CD-31 that could accelerate angiogenesis, which eventually could accelerate healing of the gastric ulcers. CONCLUSIONS: The effect of mangosteen pericap's extract and xanthone can improve healing of gastric ulcers by increasing nitric oxide and also can increase angiogenesis as shown by the increase of VEGF and CD-31 expressions.

204-15

Authors: Djanggan Sargowo, Adeodatus Yuda Handaya, Mohammad Aris Widodo, Diana Lyrawati, Askandar Tjokroprawiro

BACKGROUND: Diabetic micro and macroangiophathy lead to the incident of diabetic foot ulcers characterized by an increased number of circulating endothelial cells (CECs) and decreased function of endothelial progenitor cells (EPCs). This fact is correlated with ischemia and diabetic wound healing failure. Aloe vera gel is known to be able to stimulate vascular endothelial growth factor (VEGF) expression and activity by enhancing nitric oxide (NO) production as a result of nitric oxide synthase (NOS) enzyme activity. Aloe vera is a potential target to enhancing angiogenesis in wound healing. OBJECTIVE: The objective of this study was to explore the major role of Aloe vera gel in wound healing of diabetic ulcers by increasing the level of EPCs, VEGF, and endothelial nitric oxide synthase (eNOS), as well as by reducing the level of CECs involved in angiogenesis process of diabetic ulcers healing. METHODS: The experimental groups was divided into five subgroups consisting of non diabetic wistar rats, diabetic rats without oral administration of aloe gel, and treatment subgroup (diabetic rats) with 30, 60 and 120 mg/day of aloe gel doses for 14 days. All subgroups were wounded and daily observation was done on the wounds areas. Measurement of the number of EPCs (CD34), and CECs (CD45 and CD146) was done by flowcytometry, followed by measurement of VEGF and eNOS expression on dermal tissue by immunohistochemical method on day 0 and day 14 after treatment. The quantitative data were analyzed by One-Way ANOVA and Linear Regression, with a cofidence interval 5% and significance level (p<0.05) using SPSS 16 software to compare the difference and correlation between wound diameters, number of EPCs and CECs as well as the levels of VEGF and eNOS. RESULTS: The results of this study showed that aloe gel oral treatment in diabetic wistar rats was able to accelerate the wound healing process. It was shown by significant reduction of wound diameter (0.27±0.02); the increased number of CECs (0.42±0.57), respectively (p<0.05). On the other hand, the wound diameter and eNOS indicators showed significant differences at the dose of 60 mg, while the number of EPCs and CECs and the level of VEGF showed significantly different results at a dose of 120 mg. Aloe gel oral therapy showed a positive indication of wound healing acceleration at the optimum dose range 60-120 mg a day. CONCLUSIONS: Aloe gel is potential to be a herbal therapy candidate for diabetic wound healing through enhancing EPCs homing, decreasing the CECs number, and stimulating the increase of VEGF and eNOS levels,hence proving to be a dominant factor in the angiogenesis process.

93-109

Authors: Anna Meiliana, Andi Wijaya

BACKGROUND: The ability to reprogram mature cells to an embryonic-like state by nuclear transfer or by inducing the expression of key transcription factors has provided us with critical opportunities to linearly map the epigenetic parameters that are essential for attaining pluripotency. CONTENT: Epigenetic reprogramming describes a switch in gene expression of one kind of cell to that of another unrelated cell type. Early studies in frog cloning provided some of the first experimental evidence for reprogramming. Subsequent procedures included mammalian somatic cell nuclear transfer, cell fusion, induction of pluripotency by ectopic gene expression, and direct reprogramming. Through these methods it becomes possible to derive one kind of specialized cell (such as a brain cell) from another, more accessible tissue, such as skin in the same individual. This has potential applications for cell replacement without the immunosuppression treatments commonly required when cells are transferred between genetically different individuals. SUMMARY: Reprogramming with transcription factors offers tremendous promise for the future development of patient-specific pluripotent cells and for studies of human disease. The identification of optimized protocols for the differentiation of iPS cells and ES cells into multiple functional cell types in vitro and their proper engraftment in vivo will be challenged in the coming years. Given that the first small molecule approaches aimed at activating pluripotency genes have already been devised and that murine iPS cells have recently been derived by using non-integrative transient expression strategies of the reprogramming factors, we expect that human iPS cells without permanent genetic alterations will soon be generated.

116-21

Authors: Hera Yuliana Intantri, Andi Wijaya, Ilhamjaya Patellongi

BACKGROUND: Visceral Obesity is related with chronic low grade inflammation, and is the main component of metabolic syndrome (MetS). MetS is associated with increased cardiovascular disease (CVD). Furthermore, superoxide dismutase (SOD) is correlated with insulin resistance. Several studies have reported a strong correlation between Renin Angiotensin Aldosterone System (RAAS) and CVD, but the association of Aldosterone, Plasma Renin Activity (PRA) and SOD with inflammation, insulin resistance and MetS have not been fully elucidated. The aim of this study was to investigate the correlation of Aldosterone, PRA, and SOD with inflammation (high sensitivity c-reactive protein/hsCRP) and insulin resistance (homeostasis model assessment-insulin resistance/HOMA-IR) in adult men with central obesity. METHODS: This was a cross-sectional study, which was carried out on 80 male subjects with central obesity who were divided into 2 groups: the group of subjects who had fulfilled the MetS criteria and the other group of subjects who did not. After an overnight fasting, blood pressure (BP) was measured on all subjects and laboratory examinations were done for measurement of the concentration of fasting glucose, high density lipoprotein cholesterol (HDL-C), triglyceride, hsCRP, insulin, aldosterone, PRA, and SOD. RESULTS: We found aldosterone had positive correlation with PRA (r = 0.389; p < 0.001) and triglycerides (r =0.234; p=0.036). PRA had positive correlation with SOD (r=0.220; p=0.05) and HDL-C (r=0.273; p=0.014), but not with hsCRP (r=-0.044; p=0.696) and HOMA-IR (r=0.168 p=0.136). PRA correlated with HOMA-IR in MetS (r=0.471; p=0.01). Aldosterone and PRA were correlated with diastolic pressure in those with hypertension (r=0.680; p=0.003 and r=0.608; p=0.01). CONCLUSIONS: There is no direct correlation between aldosterone or SOD and Insulin resistance, and inflammation in men with central obesity. The correlation between PRA and MetS might be through insulin resistance mechanism. We found significant correlation between PRA, HDL-C and SOD. Increased aldosterone was ccorrelated with elevated triglycerides, thus possibly increasing the risk of MetS.

127-32

Authors: Winni Agustiani, Mansyur Arif, Ilhamjaya Patellongi

BACKGROUND: Inflammation in the vascular wall plays an important role in the pathogenesis of atherosclerosis. Current studies have shown that increase of systemic inflammatory marker like the acute phase component C-reactive protein (CRP) are associated with an unfavorable progression of disease and an increased risk for acute cardiovascular events. Recently, a close association of Metabolic Syndrome (MetS) with hemostatic abnormalities has been reported. Among hemostatic abnormalities, an increase in plasminogen activator inhibitor (PAI)-1, a strong inhibitor of fibrinolysis, is considered a core feature of MetS. High PAI-1 concentrations may be associated with thrombus formation, also causing cardiovascular events. Therefore, we investigated the association between markers for chronic inflammation (CRP) and the markers of fibrinolytic impairment (PAI-1, PAP, TAFI) in subjects with central obesity. METHODS: This was a cross-sectional study in 80 male Indonesian subjects, aged 30-60 years old with central obesity, conducted from January to March 2008 in Bandung. RESULTS: The study results showed that there was a difference of PAI-1 levels between MetS and Non-MetS group. There were significant correlations between hsCRP and PAI-1 (r=0.252, p=0.024 ), hsCRP and PAP (r=0.253, p=0.024), and also between PAI-1 and PAP (r=-0.239, p=0.033 ) respectively. But, no correlation found between hsCRP and TAFI. CONCLUSIONS: There was correlation between inflammation and fibrinolysis impairment on central obesity. Concentrations oh hsCRP, PAI-1 and TAFI were significantly higher in MetS.

138-42

Authors: Ferry Sandra, Roya Khosravi-Far

BACKGROUND: Survivin, a bifunctional protein, acts as suppressor of apoptosis and has an essential role in mitosis. Survivin is physically phosphorylated on Thr34, and other important sites such as Thr117, Ser20, Thr48 and Ser81. Our previous report has shown that Ser81 of survivin plays role in cytoprotection. In order to investigate the underlying mechanism, all motifs with medium stringency were scanned. We found that site of survivin at Ser81 was correlated to PKA, which is well reported to many cell signal machineries, including cell survival. Therefore, we focused our current investigation in finding possible correlation and interaction between survivin’s Ser81 site and PKA. METHODS: Wild-type survivin (Survivin), antisense survivin (Survivin-AS), mutated-survivin and mutated-survivin Ser81Ala (Survivin-S81A) were constructed. Each retroviral product was produced. Some cell lysates were prepared and immunoprecipitated. For analysis, we performed immunoblotting and PKA’s activity assays. RESULTS: In our current results, phosphorylated-PKA was correlated with survivin. Infection of survivin could lead to acceleration of PKA’s activity in a viral particle dependent manner. This positive back loop induction by survivin was shown to be correlated to Ser81 site, since survivin-mediated PKA activity was not resulted by mutated form of survivin at Ser81 to nonphosphorylatable Ala (S81A). CONCLUSIONS: Our results suggested a possible back loop of survivin to activate PKA, and Ser81 could be an important site to mediate the survivin-PKA back loop signaling. Survivin-induced activation of PKA might be related to cytoprotection.

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.