The Indonesian Biomedical Journal

94-125

Authors: Irma Ruslina Defi, Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: Fibronectin type III domain-containing protein 5 (FNDC5), or also known as irisin, has been identified for two decades but almost completely disregarded for 10 years. It is present in skeletal muscle, heart, and brain, and in reaction to exercise can transform white adipose tissue into brown. Since then, irisin has gained a lot of attention for its potencies in treating metabolic disorders. In this review article, the potential future of irisin especially on metabolism and aging process will be discussed. CONTENT: Sedentary lifestyle is acknowledged as risk factor for type 2 diabetes mellitus, obesity, immune system issues, asthma, and neurological or heart illness. Irisin is secreted by muscle cells when exercising, produced after the proteolytic cleavage of FNDC5 protein. Irisin has positive impacts on maintaining physiological balance including reducing inflammation, keeping the bone homeostasis, as well as influencing metabolic processes and the neurological system function. Due to these many and advantageous characteristics, irisin could be a possible choice for preventing and managing disorders associated with modern society, and finding the agents to increase irisin can be beneficial. SUMMARY: Irisin offers a fresh potential basis for kinesitherapy and shows promise as a therapeutic target due to its various biological activities. Irisin pathway can be activated through dietary changes, the use of natural substances and drugs and can interact with this signalling pathway which involved peroxisome proliferator-activated receptor-γ coactivator 1α (PGC-1α) and uncoupling protein mRNA 1 (UCP1) to resolve obesity and its metabolic comorbidities.

135-43

Authors: Primariadewi Rustamadji, Elvan Wiyarta, Meike Pramono, Sinta Chaira Maulanisa

BACKGROUND: Neoadjuvant chemotherapy (NAC) plays a crucial role in the management of invasive breast cancer with no special type (IBC-NST), with the immune system's response to cancer heavily relying on the dynamics between tumor-infiltrating lymphocytes (TILs) and cancer cells. In this study, the differential effects of anthracycline-based NAC on stromal and intratumoral foxhead box P3 (FOXP3+) TILs expressions were specifically examined. METHODS: In this cross-sectional study, 32 IBC-NST samples were evaluated for pre- and post-NAC FOXP3+ TIL expression as well as the changes of FOXP3+ TIL expression. Comprehensive data collection regarding subjects' age, tumor size, grade, lymphovascular invasion, regional lymph node metastasis, and receptor status were conducted. Immunohistochemistry was utilized to quantify FOXP3+ TILs. The stromal, intratumoral and total FOXP3+ TILs expression were then analyzed. RESULTS: Significant reductions in FOXP3+ TIL expression post-NAC were observed, with stromal FOXP3+ TILs showing a median decrease of 3.6 units in subjetcs aged ≥50 years (p=0.013) and a median decrease of 13.2 units in subjects with tumors ≥5 cm after NAC (p=0.006). In contrast, intratumoral FOXP3+ TILs remained relatively stable, with minor changes. The total FOXP3+ TIL expression, combining stromal and intratumoral components, was significantly decreased with a median of 13.0 units decreased to 5.3 units (p<0.001). CONCLUSION: This study highlights the significant reduction in stromal FOXP3+ TIL expression after NAC treatment in IBC-NST subjects, in contrast to the relatively stable intratumoral FOXP3+ TILs. Understanding these differences may guide future therapeutic strategies and improve treatment outcomes for IBC-NST.

152-61

Authors: Sri Yani, Salim Harris, Ahmad Aulia Jusuf, Roman Goenarjo, Feda Anisah Makkiyah, Rifqi Sabita, Nurhadi Ibrahim

BACKGROUND: Stroke is one of the major causes of disability in the world. High-intensity interval training (HIIT) is known as a novel treatment to promote stroke recovery. However, the results differ in their effects on irisin, which is a regulator of brain-delivered neurotrophic factor (BDNF). Therefore, this study was conducted to investigate the effect of HIIT on BDNF and irisin levels in a rat model of ischemic stroke with middle cerebral artery occlusion (MCAO) induction on recovery motor function. METHODS: Rats were categorized into 4 groups: sham, MCAO, MCAO+moderate-intensity interval training (MIIT), and MCAO+HIIT. MCAO induction was performed to create the ischemic stroke rats model. The motor function was assessed through rotarod and footprint tests. Blood samples were obtained 6 days before MCAO and 14 days after MCAO to examine BDNF and irisin levels with enzyme-linked immunosorbent assay (ELISA). Brain tissue samples were collected 14 days after MCAO for histopathological examination of cortical tissue with hematoxylin-eosin (HE) staining. RESULTS: Rats in the MCAO+HIIT group exhibited an enhanced ability to walk on the rotarod (p=0.016). The stride-length hind paw right in the MCAO+HIIT group demonstrated a noteworthy increase in comparison to baseline value (p=0.036), and the stride-length fore paw right showed a significant increase in both the MCAO+HIIT (p=0.036) and MCAO groups (p=0.034). BDNF significantly improved in the MCAO+MIIT (p=0.043) and MCAO+HIIT groups (p=0.018). The irisin level only showed a significant enhancement in the MCAO+HIIT group (p=0.018). CONCLUSION: HIIT increased motor function, while BDNF level increased with HIIT and MIIT intervention. This preclinical research is useful for supporting the recovery of stroke patients by HIIT intervention.

172-9

Authors: Merci Monica br Pasaribu, Suzanna Immanuel, Damayanti Rusli Sjarif, Ina Susianti Timan, Safarina G. Malik, Muchtaruddin Mansyur, Ernawati Simanjuntak

BACKGROUND: Amino acids are one of the essential metabolites, especially the 20 amino acids that are preserved as the building blocks of protein. Alterations in amino acid concentrations are related to disease such as inborn error of metabolism, cancer, as well as nutritional status. Hence, it is necessary to define reference values of 20 plasma-free amino acids for Indonesian toddlers and to establish a robust measurement technique using chromatography with tandem mass spectrometry (MS). METHODS: This study was a cross-sectional preliminary study to establish reference values. The sample was prepared by mixing plasma with 20% sulfosalicylic acid. Plasma-free amino acids were measured with high-performance liquid chromatography (HPLC) non-derivatization technique using column XTerra for chromatographic separation coupled with tandem MS. Amino acids reference values were taken from 101 healthy Indonesian toddlers aged 1-3 years old. Since amino acids data were not Gaussian distributed, the lower and upper of the reference value was established from the 5th percentile and the 95th percentile, respectively. RESULTS: Analysis for 20 amino acids was validated. The accuracy ranged from 90.53-105.39% and the precision ranged from 0.06-3.80%. The limit of detection range was 1-2 nmol/mL, and the limit of quantification range was 2-4 nmol/mL. The result was linear, with R2 higher than 0.998. There was no significant difference between boys and girls for all amino acids except for glycine. CONCLUSION: HPLC with tandem MS method can be used to evaluate amino acids in clinical practice. The reference values obtained are specific for aged 1-3 years old from urban areas in Indonesia. The study suggests that for each population, the reference values for amino acids should be established.

189-98

Authors: Ermin Rachmawati, Maulidya Machdaniar, Larasati Sekar Kinasih, Djanggan Sargowo, Brillian Putra Syarifuddin, Fayza Cinta Rahmadina Putri, Mahrus Ismail

BACKGROUND: Non-alcoholic fatty liver disease (NAFLD), characterized by inflammation and steatosis, is one of the metabolic problems that trigger serious complications. Statin is usually used for NAFLD therapy, however, statin intolerance and resistance reduce the effectiveness of NAFLD treatment. Therefore, this study was conducted to develop the phytopharmaca against NAFLD by elucidating the anti-inflammatory and steatosis suppression of vegetable spices fermentation extract (VSFE) in rabbits induced with high-fat diet (HFD). METHODS: Twenty New Zealand White rabbits were determined into 4 groups; which were groups receiving standard diet (SD), HFD, HFD and 100 mg/kgBW VSFE, as well as HFD and 200 mg/kgBW VSFE. mRNA expressions of tumor necrosis factor alpha (TNFα) and CCAAT/enhancer-binding protein alpha (C/EBPα) were evaluated. The TNFα concentration was quantified after protein and RNA isolation. Inflammation and steatosis were observed in liver hematoxylin-eosin-stained preparate using microscope. RESULTS: Macrophage cells and hepatic steatosis showed significant decrease in a drug dependent manner. An effective dose of 200 mg/kgBW decreased C/EBPα and TNFα expression to 0.45±0.32 and 0.72±0.29 (p=0.013 and p=0.002, compared to the HFD group), respectively. TNFα inflammatory cytokine concentrations also responded to the administration of 100 mg/kgBW and 200 mg/kgBW doses, with the value of 31.72±10.40 mg/dL and 48.35±7.15 mg/dL (p=0.009 and p=0.002, compared to the HFD group), respectively. CONCLUSION: VSFE might prevent NAFLD by inhibit steatosis and inflammation through suppression of C/EBPα expression and TNFα expression.

1-22

Authors: Anna Meiliana, Nurrani Mustika Dewi, Andi Wijaya

BACKGROUND: World Health Organization has reported four million people die every year due to obesity comorbidity, and the prevalence of obesity is keep increasing, especially after COVID-19. Obesity has been defined as a chronic disease involving adipose tissue dysfunction which leads to metabolic diseases and psychosocial consequences. The review article will highlight some recent researches regarding the new conceptual framework that integrates both metabolic drives, as well as to summarize the numerous discussions about the current understanding of hypothalamic control of food intake and energy homeostasis. CONTENT: Obesity apparently is not simply regulated only by food and exercise. Hypothalamus takes part in controlling energy intake and expenditure via appetite regulation. Hedonic control in cortical and subcortical brain areas process cognitive, reward, information, and executive function. Managing metabolic adaptation, browning the white adipose tissue, and preserving lean mass can be another strategy to safely manage obesity. SUMMARY: Obesity need to be managed in a multimodal strategy including neurophysiology and physiology approach, together with environment support. Thus, a weight regain can be prevented. Commitment from both scientific and regulation point of view can shed a light to eradicate obesity.

31-9

Authors: Edy Fachrial, Juliana Lina, Harmileni, Sari Anggraini, Widya Yanti Sihotang

BACKGROUND: Type 2 diabetes mellitus (T2DM) cases tend to rise throughout the year in all parts of the world. The α-glucosidase inhibitors are frequently used to both prevent and treat T2DM. This α-glucosidase inhibitor activity is seen in some probiotic species, and a certain strain of Pediococcus acidilactici exhibits promising characteristics as an α-glucosidase inhibitor. This study was conducted to assess the hypoglycemic activity and safety of P. acidilactici strain DNH16 (PADNH16) in experimental rats with T2DM induced by streptozotocin. METHODS: The experiment was conducted in vivo using Wistar rats. Acarbose was employed as a positive control, and Lactobacillus casei was used as a comparative. For 24 days, post-prandial blood glucose levels were assessed every 3 days, followed by serum biochemical levels measurement of liver, kidney, and lipid profiles. The pancreas was histopathologically examined utilizing the Hematoxyline-Eosin staining procedure. RESULTS: Administration of PADNH16 to T2DM rats lowered post-prandial blood glucose levels and gave hypoglycemic benefits comparable to acarbose and commercial probiotics. PADNH16 dosing did not affect serum glutamic oxaloacetic transaminase (SGOT), serum glutamic pyruvic transaminase (SGPT), urea, or creatinine levels, showing that PADNH16 was not hazardous to liver or kidney function. The lipid profile assessment revealed that the values, including high-density lipoprotein (HDL), low-density lipoprotein (LDL), and triglycerides, were comparable to the control group. The pancreatic histopathology revealed that injection of PADNH16 caused no alterations to pancreatic β cells. CONCLUSION: P. acidilactici isolate DNH16 has a hypoglycemic effect on T2DM rats, but does not affect liver function enzymes, kidneys, lipid profiles and does not provide significant changes in pancreatic β cells.

48-55

Authors: Muhammad Ridwan, Herlina Dimiati, Maimun Syukri, Ronny Lesmana, Lia Meuthia Zaini

BACKGROUND: Sodium glucose co-transporter-2 inhibitor (SGLT2-i), a new oral antidiabetic drug, has been recommended for its morbidity and mortality benefits in patients with heart failure. This study aimed to determine the effect of acute SGLT2-i therapy on the relative ratio expression of microRNA-21 (miR-21), transforming growth factor-β1 (TGF-β1), and matrix metalloproteinase-2 (MMP-2) in the process of cardiac fibrosis in the hyperglycemia Wistar rat models compared to biguanide (metformin) therapy. METHODS: We used Streptozotocin (STZ) to induce hyperglycemia in Wistar rats (n=31), randomly divided into four groups: negative control (NC, n=4), positive control (PC, n=10), hyperglycemia plus metformin (M, n=8), and hyperglycemia plus empagliflozin (E, n=9). After seven weeks, the rats were sacrificed and the heart tissue was taken for microRNA and messenger RNA (mRNA) extraction, followed by reverse transcription quantitative real time polymerase chain reaction (RT-qPCR) examination. The data was analyzed using One-way ANOVA. RESULTS: Results showed a decreasing trend in the gene expression relative ratio of miR-21 (1.0 vs. 1.9; p=0.079) and TGF-β1 (0.9 vs. 3.2; p=0.145), but a significant increase in MMP-2 gene expression (1.3 vs. 0.7; p=0.002) in the SGLT2-i (empagliflozin) vs. biguanide (metformin) groups. CONCLUSION: Empagliflozin administration may play a significant role in preventing the occurrence of cardiac fibrosis in hyperglycemia.

66-71

Authors: Ryco Giftyan Ardika, Bernardus Parish Budiono, Nyoman Suci Widiastiti, Nani Maharani, Neni Susilaningsih, Ferry Sandra

BACKGROUND: Microcirculation and cellular disturbances caused by sepsis might trigger significant intestinal damage. Andrographis paniculata extract decreases inflammatory intestinal epithelial cells with its role as an antiparasitic and anti-inflammatory agent. However, A. paniculata extract’s effect on sepsis have not been commonly studied, especially in the intestinal tissues. Therefore, this study was conducted to determine A. paniculata leaves extract (APLE) effect in sepsis-induced intestinal tissues of rats by examining the expression of inflammatory cytokines involved in sepsis, namely tumor necrosis factor (TNF)-α and Caspase-3. METHODS: Rats were divided into five groups; two groups received no pretreatment and the other three groups received 200, 400, and 500 mg/kg BW/day APLE, respectively. Three pretreated groups and one group with no pretreatment were then injected with 1 mg/200 g BW lipopolysaccharides (LPS) intraperitoneally to create septic rat models. Three days after the LPS-induction, rats were euthanized and the expression of TNF-α and Caspase-3 were assessed based on the immunohistochemical staining of rats’ intestinal tissues. RESULTS: Compared with NaCl (sham), LPS significantly (p<0.001) induced TNF-α expression from 6.60±1.36 to 25.37±1.74. Pretreatment of 200, 400, and 500 mg/kg BW/day APLE could significantly (p<0.001) inhibit the LPS-induced TNF-α expression (18.82±1.36, 11.45±1.18, and 6.89±1.90, respectively). Similar with TNF-α, compared with NaCl (sham), LPS significantly (p<0.001) induced Caspase-3 expression from 6.92±1.66 to 23.59±2.25. Pretreatment of 200, 400, and 500 mg/kg BW/day APLE could significantly (p<0.001) inhibit the LPS-induced Caspase-3 expression (17.47±1.68, 12.99±1.51, and 5.59±1.51, respectively). CONCLUSION: The pretreatment of APLE could inhibit the LPS-induced TNF-α and Caspase-3 expression, therefore APLE could be suggested as a potential sepsis-preventing agent.

79-87

Authors: Ujang Saeful Hikmat, Ani Retno Prijanti, Heri Wibowo, Indriyanti Rafi Sukmawati, Dicky Levenus Tahapary

BACKGROUND: Type 2 Diabetes (T2D) impairs the innate immune system including monocytes. Monocytes are divided into two subgroups depending on the expression of cluster of differentiation (CD)14 and CD16 receptors, namely CD14+CD16- and CD14+CD16+. CD14+CD16+ are proinflammatory monocytes and develop into M1 type macrophages, which contribute to foam cell production, a risk factor for cardiovascular disease (CVD). Therefore, it is important to determine the influence of T2D conditions on changes in monocyte subsets and whether these changes correlate with CVD risk markers. METHODS: Peripheral blood mononuclear cell (PBMC) was obtained from 10 T2D subjects and 10 healthy donors. Subsequently, PBMC was incubated for 24 hours with and without 10 mL lipopolysaccharide. Flow cytometry was used to evaluate CD14 and CD16 expression, while multiplex immunoassays were applied to measure interleukin (IL)-1b and IL-10 concentrations in supernatants. RESULTS: In T2D, the percentage of CD14+CD16+ monocytes increased (p=0.07), and an increase in CD14+CD16+ monocytes more than 6.8% was linked with CVD risk markers (r=10.146, p=0.002). Meanwhile, inflammatory mediators released by monocytes shown an increase in IL-1b (p=0.041) but not in IL-10 (p=0.082) in T2D subjects. Fasting blood glucose levels were also found to be substantially linked with an increase in CD14+CD16+ monocytes (r=0.530, p=0.016). CONCLUSION: T2D patients had a higher percentage of CD14+CD16+ monocytes and IL-1b levels than healthy donors. An increase in CD14+CD16+ monocytes above 6.8% associated with CVD risk markers in T2D patients.