The Indonesian Biomedical Journal

226-42

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Metabolism impairment in obese condition usually initially triggered by inflammation and insulin signaling impairment. The involvement of metabolites, including lipids, amino acids, and ketone bodies, in altering insulin sensitivity has been revealed after massive data sets were provided by the studies regarding metabolomics and lipidomics. CONTENT: Metabolites were now understood to serve more than just the metabolism products, but also as active signaling molecules including in insulin and immunological actions. Different lipid metabolites can serve as signaling molecules to induce insulin resistance of sensitivity through a similar pathway, and impact on the inflammation status. Branched Chain Amino Acids (BCAA) and many amino acids have been correlated with mitochondrial dysfunction and insulin impairment. Ketogenic diet, supplementation and microbiota transplantation become the current strategies to set a preferable metabolites composition to modulate insulin sensitivity. SUMMARY: Thousands of metabolites can now be measured using technical and bioinformatics developments. Different types of amino acids, fatty acids, and bile acids are being studied in relation to altered metabolic states, particularly obesity and type 2 diabetes mellitus. A thorough knowledge of the metabolic changes that contribute to insulin resistance might lead to the discovery of new targets for enhancing insulin sensitivity and preventing and treating many metabolic disorders.

252-60

Authors: Astrid Feinisa Khairani, Nur Atik, Putri Halleyana Adrikni Rahman, Enny Rohmawaty, Cynthia Noviyanti, Resti Santika, Jose Arimathea, Widad Aghnia Shalannandia

BACKGROUND: Purple sweet potato yogurt (PSPY) is a funtional food which is rich in anthocyanin and probiotics. However, the currently available data on its potentially protective effect on anthropometry, lipid metabolism, oxidative stress, and pro-inflammatory markers is very minimal, especially in mice. This study was performed to investigate those effects on balb/c mice models (Mus musculus) given a high-fat diet (HFD). METHOD: Balb/c mice were treated with or without standard diet, HFD, ethanol extract, yogurt, and PSPY according to the group. The changes of anthropometry were analyzed using Lee Index. After three months, the interscapular brown adipose tissue (iBAT) was morphologically observed with hematoxylin and eosin (H&E) staining. The blood serum was used for evaluation using cholesterol oxidase-peroxidase aminoantypirin (CHOD-PAP) for lipid profile, enzyme-linked immunoassay (ELISA) for tumor necrosis factor (TNF)-α and interleukin (IL)-6, and thiobarbituric acid reactive substance (TBARS) procedure for malondialdehyde (MDA). RESULT: Lee Index revealed a decrease in time (p<0.0001). The PSPY group showed a decrease in iBAT weight (p<0.05), lipid profiles including LDL (p<0.05) and total cholesterol (p>0.05), TNF-α and IL-6 (p>0.05), and MDA (p>0.05). Adipocytes’ density showed a significant increase (p=0.001). CONCLUSION: This research finding indicates that PSPY affects lipid metabolism and has a potential protective effect of reducing systemic inflammation and oxidative stress.

269-75

Authors: Budi Susetio Pikir, Andrianto Andrianto, I Gde Rurus Suryawan, Hanestya Oky Hermawan, Dian Paramita Kartikasari, Primasitha Maharany Harsoyo

BACKGROUND: Transdifferentiation is a method to provide cells sources for cellular cardiomyoplasty. CD34+ cells are potential cells sources because these cells can differentiate into cardiomyocytes through several mechanisms. MicroRNA (miR-1) is known to have the ability to inhibit the expression of histone deacetylase 4 (HDAC4). HDAC4 is a gene that essentially contributes in cardiomyocytes differentiation. However, the study reporting an evidence that miR-1 can induce transdifferentiation of CD34+ peripheral blood cells into mature cardiomyocytes is limited. METHODS: CD34+ cells were taken from peripheral blood and isolated using a magnetic-activated cell sorting (MACS) method in vitro. Mature mimics of miR-1 were transfected into isolated CD34+ cells and then incubated for 48 hours for quantification of HDAC4 mRNA using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). On the fifth day after miR-1 transfection, cardiomyocyte-like cells were identified based on their morphology and cardiac troponin expression using immunocytochemistry. RESULTS: Transfection of miR-1 in CD34+ isolated cells decreased HDAC4 gene expression by -0.54 fold at second day and caused a significant increase in percentage of cardiac troponin positive cells (median: 31.34; p<0.05) at fifth-day post-transfection. The efficiency of transdifferentiation was 32%. The miR-1 transfection had a significant negative relationship with HDAC4 gene expression (B=-1.000; p=0.001). HDAC4 gene expression had a negative and significant relationship with the percentage of cardiac troponin-positive cells (B=-0.701; p=0.001). CONCLUSION: This study suggests that miR-1 can induce transdifferentiation of peripheral blood CD34+ cells into cardiomyocytes-like cells by decreasing HDAC4 gene expression.

282-8

Authors: Edhyana Sahiratmadja, Ani Melani Maskoen, Lelani Reniarti, Delita Prihatni

BACKGROUND: Being located in the global thalassemia belt area, Indonesia is estimated harboring about 10% thalassemia carriers; however, screening program is still diversely scattered across the country. Numerous erythrocyte indices have been introduced to help identifying thalassemia carriers with contradictory results. Therefore, this study had compared the use of mean corpuscular volume (MCV) and/or mean corpuscular hemoglobin (MCH) values and the most erythrocyte indices used in Indonesia which were Mentzer Index (MI) and Shine & Lal Index (SLI), as a first attempt in a mass screening for β-thalassemia carrier. METHODS: This was a retrospective study, evaluating laboratory data from family members of thalassemia major subjects. The sensitivity and specificity of MI and SLI were calculated. HbA2 >3.5% was used as a golden standard for β-thalassemia carrier and DNA examination was conducted to confirm β-globin mutation. RESULTS: Out of 160, 28.8% of the subjects had low Hb concentration. Interestingly, 79.4% of the subjects had low MCV and/or MCH with or without low Hb concentration. In this study, specificity and sensitivity of MI were 82.2% and 83.8%, whereas of SLI were 96% and 40.5%, respectively. Low MCV and/or MCH had covered IVS1nt5 and Cd26 mutation at β-globin gene; whereas MI and SLI had missed some samples, leading to false negative of thalassemia carrier results, when using MI or SLI only. CONCLUSION: MCV<80 fl and/or MCH<27 pg is the best first round mass screening method for β-thalassemia carrier in a limited facility area. However, Hb electrophoresis should be gradually installed regionally in various places wherever possible, as well as DNA analysis to confirm the mutation for an optimal carrier diagnosis.

294-302

Authors: Hendri Susilo, Budi Susetyo Pikir, Mochammad Thaha, Mochamad Yusuf Alsagaff, Satriyo Dwi Suryantoro, Citrawati Dyah Kencono Wungu, David Setyo Budi, Laurentius Andre, Cennikon Pakpahan

BACKGROUND: Cardiovascular disease (CVD) is the leading cause of death in patients with chronic kidney disease (CKD). Klotho expression was reduced in patients with CKD, leading to vascular calcification, endothelial dysfunction, and atherosclerosis. We investigated the role of the klotho G395A gene polymorphism and plasma klotho level in the ten-year risk of atherosclerotic cardiovascular disease (ASCVD) and CVD mortality in CKD patients. METHODS: We used the PCR-CTPP assay method to genotype klotho G395A single nucleotide polymorphism (SNP) in 72 non-dialysis CKD patients. The klotho level was determined using the enzyme-linked immunoassay (ELISA) method. Path analysis was used to determine the relationship between the klotho G395A SNP, plasma klotho level, ASCVD risk score, and CVD mortality risk score. RESULTS: Our results showed that the GA genotype had lower plasma klotho levels than the GG genotype (path coefficient=-0.185, p=0.000). There was a significant negative correlation between plasma klotho level and the ASCVD risk score (r=-0.243, p=0.040), but no significant correlation was found between plasma klotho level and the CVD mortality risk score (r=-0.145, p=0.225). Path analysis showed that plasma klotho level had a significant negative direct effect on ASCVD risk score (path coefficient=-0.272, p=0.000) and an indirect effect on CVD mortality risk score (path coefficient=0.187, p=0.005). CONCLUSION: Klotho G395A SNP might reduce lower plasma klotho levels, which increased ASCVD and CVD mortality risk scores in non-dialysis CKD patients. However, other risk factors such as age, CKD stages, hypertension, and smoking should be taken into consideration. Therefore, large-scale genetic association studies with adjusted variables could be conducted in various ethnic groups for a more robust result.

309-15

Authors: Ratih Tri Kusuma Dewi, Dhani Redhono, Agung Susanto, Diding Heri Prasetyo, Evi Nurhayatun, Ida Nurwati, Bambang Purwanto

BACKGROUND: Anthrax is a zoonotic disease caused by Bacillus anthracis, whose endospores stimulate the release of pro-inflammatory cytokines and promote oxidative stress. Propolis, a natural resource that can be found in Indonesia, has been proven to have anti-apoptotic and antioxidant role which might be a potential adjuvant therapy for anthrax treatment. Hence in this study we aimed to investigate effect of propolis as an anti-inflammatory, anti-apoptotic, and antioxidant in anthrax rats model by examining the level of tumor necrosis factor (TNF)-α, caspase-3, and malondialdehyde (MDA), respectively. METHODS: This was an experimental post-test only study with 40 male rats weighed 180-200 g that were induced by anthrax spores injected subcutaneously. The rats were divided into one control positive group and four intervention groups that were administered with 200 mg/kgBW propolis extract for 7 to 14 days. The levels of serum TNF-α, caspase-3, and MDA were measured using enzyme-linked immunosorbent assay (ELISA) and analyzed with bivariate analysis. RESULTS: TNF-α, caspase-3, and MDA level were found lower in anthrax rats model given ethanol extract of propolis than the control group. The lowest concentration of TNF-α value was found in group administered with propolis extract 200 mg/kgBW for 7 days before the anthrax induction (6.136±0.205 pg/mL). The results were similar to the MDA serum and caspase-3 which were the lowest when the propolis was administered 7 days earlier (1.893±0.188 nmol/mL and 2.040±0.067 ng/mL). There was significant difference in the TNF-α, caspase-3, and MDA serum levels (p≤0.001) compared to control group. CONCLUSION: Propolis has anti-apoptotic and antioxidant effects which can be used as complementary therapy in anthrax infection.

323-8

Authors: Ferry Sandra, Muhammad Ihsan Rizal, Ayasha Hajjar Audreyandra Wahid, Monica Andajana, Maria Celinna

BACKGROUND: Caffeic acid has been reported to induce apoptosis in MG-63 osteosarcoma cells via caspases activation. However, apoptotic pathway that is involved in the caffeic acid-induced apoptosis is still unclear. Present study aimed to investigate the role of cytochrome c (Cyt c) release and BH3-interacting death (Bid) activation in caffeic acid-induced apoptosis in MG-63 osteosarcoma cells. METHODS: MG-63 cells were cultured, pretreated with/without Z-VAD FMK and treated with/without 10 μg/mL caffeic acid. Treated MG-63 cells were then lysed, homogenized, and processed further to prepare cell lysate and mitochondrial fraction. Immunoblotting method was used to measure the amount of Bid and truncated Bid (t-Bid) as well as mitochondrial and cytosolic Cyt c. RESULTS: The amount of Bid and mitochondrial Cyt c in MG-63 cells decreased in a time-dependent manner, while the amount of t-Bid and cytosolic Cyt c increased in a time-dependent manner. By pretreatment of 100 μM Z-VAD-FMK for 2 h, the amount of Bid and mitochondrial Cyt c was significantly higher, while the amount of t-Bid and cytosolic Cyt c was significantly lower after caffeic acid treatment for 6 and 12 h compared to MG-63 cells that were not pretreated. CONCLUSION: Caffeic acid could induce Cyt c release through the activation of Bid in MG-63 osteosarcoma cells.