The Indonesian Biomedical Journal

296-302

Authors: Maki Yoshikawa, Masatoshi Oshita, Tomomi Gotoh, Yukina Tawa, Katsuaki Dan

BACKGROUND: Stem cell culture medium is garnering attention because it comprises growth factors and exosomes secreted by stem cells. In this study, stem cell culture medium was subjected to electric field stimulation using the cells alive system (CAS) to determine the antiaging effects of this medium on dermal fibroblasts. METHODS: The medium was added to the monolayer cultures of human fibroblasts with or without 100 μg/mL advanced glycation end products (AGE), and the cells were incubated for 4 hours. Total RNA was extracted, and mRNA levels of several AGE receptors and heat shock proteins were measured using quantitative reverse transcriptase-polymerase chain rection (qRT-PCR). RESULTS: Similar amounts of exosomes were extracted from the supernatants of stem cells cultured with (CAS+) and without (CAS−) electric field stimulation. The results demonstrated that CAS+ had a higher oxygen radical absorbance capacity than CAS−, indicating its high antioxidant activity. Furthermore, the expression of domain-containing link scavenger receptor-1 mRNA, which is involved in the degradation of cell surface receptors for AGE under glycation stress, significantly increased. The electric field stimulation also increased the expression of several heat shock proteins, which play important roles in cellular responses generated against glycation stress. The electric field stimulation of the stem cell culture medium effectively promoted the endocytosis and degradation of AGE, thereby exerting antioxidant effects and inducing glycation stress. CONCLUSION: Electric field stimulation of stem cell culture medium enhanced the expression of Hsp and its antioxidant activity in fibroblasts.

310-5

Authors: Faisal Faisal, Nur Rochmah, Muhammad Faizi, Novina Novina, Erni Nuraeni

BACKGROUND: Type 1 diabetes (T1D) mostly occurs due to the destruction of pancreatic beta cells due to autoimmune processes. Diagnosis of T1D can be established by examining the c-peptide levels and the markers of pancreatic autoantibodies, including glutamic acid decarboxylase 65 autoantibodies (GAD-65). However, the association between c-peptide and anti-GAD-65 toward patients’ clinical manifestations needs to be further explored. Hence, the aim of current study was to identify the association of anti-GAD65 with c-peptide and clinical characteristics in children with T1D. METHODS: Case-control study involving 47 T1D children (T1D group) and 41 healthy children (control group) younger than 18 years old was conducted. Secondary data regarding subjects’ demographic characteristics and medical history were collected from subjects, and serum blood was drawn from each subject for the anti-GAD65 and c-peptide measurement. Anti-GAD65 and c-peptide levels were measured using an Enzyme-Linked Immunosorbent Assay (ELISA) methods. RESULTS: Anti-GAD65 antibody was detected in 78.7% T1D group, while only 2.43% were detected in control group subject (p=0.0000). The c-peptide level of T1D group was 0.07±0.19 nmol/L and control group was 1.5±0.77 nmol/L (p=0.0000). The total daily dose of insulin in subjects with positive anti-GAD65 was greater than in the negative anti-GAD65 (p=0.012). The sensitivity and specificity of the anti-GAD65 were 85.4% was 66.7%, respectively. CONCLUSION: The results of this study show that anti-GAD65 was associated with total daily dose of insulin requirement in children with T1D.

322-32

Authors: Dinda Aliffia, Dinda Ayu Ramadhani, Widya Wasityastuti, Dewi Ratih Tirto Sari, Ulayatul Kustiati, Hevi Wihadmadyatami, Dwi Liliek Kusindarta

BACKGROUND: Neurodegenerative diseases are characterized by the loss of neuronal function in the nervous system. In recent years, more than 45 million people worldwide have suffered from progressive loss of memory and cognitive functions caused by Alzheimer’s disease. Ocimum sanctum is one of the medicinal plants known to have neuroprotective abilities. This study was conducted to elucidate the anti-apoptotic effects of ethanolic extract of O. sanctum (EEOS) on PC12 and SH-SY5Y cells as well as interaction between main compounds of EEOS and β-amyloid (Aβ) peptide through in silico molecular docking. METHODS: The viability of TMT-induced PC12 and SH-SY5Y cells was assessed with 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay and acridine orange/propidium iodide staining. Cell proliferation rate was measured with cell counting kit-8 (CCK-8) assay. Nuclear fragmentation was observed with Hoechst 33342 staining. Caspase -3 and -9 expressions were measured using enzyme-linked immunosorbent assay. Interactions between main compounds of EEOS and Aβ were visualized with in silico molecular docking. RESULTS: EEOS had the potential effect of maintaining cell viability, preventing the cell’s morphological changes, and inhibiting apoptosis via the caspase pathway in PC12 and SH-SY5Y cells. Meanwhile, flavonoid K, phenol, eugenol could interact with the active site of Aβ through hydrogen-bonding and hydrophobic interactions. CONCLUSION: EEOS could prevent neuronal cell apoptosis via downregulation of caspase-3 and -9. Main compounds of EEOS could interact with the active site of Aβ, and thereby might inhibit Aβ oligomerization. Thus, EEOS and its main compounds could be potential as neuroprotective agents for preventing neurodegenerative diseases.

339-46

Authors: Yudy Tjahjono, Caroline Caroline, Jusak Nugraha, Kuncoro Foe, Srikanth Karnati, Süleyman Ergün, Nico Jafet, Oryza Chrisantia, I Made Andika Bara Kusuma, Hendy Wijaya, Wuryanto Hadinugroho, Dwi Aris Agung Nugrahaningsih, Dwi Liliek Kusindarta, Yufita Ratnasari Wilianto, Senny Yesery Esar, Hevi Wihadmadyatami

BACKGROUND: Lipopolysaccharide (LPS) has been reported to increase CD4+ regulatory T-cell (CD4+ Treg) populations. Acetylsalicylic acid (ASA) has been reported to have immunomodulatory activity, but it may induce chronic gastric ulceration. Another salicylic acid-bearing compound, 2-(3-(chloromethyl)benzoyloxy)benzoic acid (3-CH2Cl), has been reported to have less gastric mucosal damage. However, the effect of 3-CH2Cl on CD4+ Tregs in LPS-induced mice is still unknown. Therefore, the present study was conducted to investigate the immunomodulatory effect of 3-CH2Cl on CD4+ T-cell and CD4+ Treg populations as well as FoxP3 expression in LPS-induced mice. METHODS: Synthesis of 3-CH2Cl was performed by mixing salicylic acid and chloromethylbenzoylchloride with the catalyzation of pyridine, acetone and heat. The 3-CH2Cl tablets were prepared using direct compression method. After intraperitoneal injection of 1 mg/kg BW LPS to mice, 60 mg/kg BW ASA or 60 mg/kg BW 3-CH2Cl was given orally for 3 days. The splenocyte was obtained through splenectomy and collagenase digestion. The population of CD4+ T-cells and CD4+ Tregs, as well as the splenic FoxP3 expression were determined using flow cytometry technique. RESULTS: CD4+ T-cell populations in mice treated with LPS and 3-CH2Cl or ASA were lower than those treated with LPS merely. Meanwhile, CD4+ Treg populations and FoxP3 expression levels in mice treated with LPS and 3-CH2Cl or ASA were higher than those treated with LPS merely. CONCLUSION: Since 3-CH2Cl could decrease CD4+ T-cell population and increase CD4+ Treg population mediated by the increase of FoxP3 expression in LPS-induced inflammation, it may act as a potential therapeutic drug to reduce inflammatory conditions.

354-61

Authors: Bantar Suntoko, Suharyo Hadisaputro, Handono Kalim, Suyanto Hadi, Ika Vemilia Warlisti

BACKGROUND: Systemic lupus erythematosus (SLE) is an autoimmune condition characterized by persistent, chronic inflammation that damages organ tissue. One of the symptoms that is often found in SLE is cognitive dysfunction. Hydroxychloroquine is recommended for the treatment of all levels of SLE. This study was conducted to prove the influence of hydroxychloroquine on improving cognitive function and inflammatory biomarkers compared to standard therapy. METHODS: The study adopted randomized controlled trial (RCT) in SLE patients with cognitive dysfunction who met the inclusion criteria. The treatment group consisted of 26 subjects who received hydroxychloroquine 200 mg/day for 8 weeks and standard therapy, while the control group consisted of 29 subjects who were given standard therapy only. Examination of Montreal Cognitive Assessment (MoCA)-INA, interleukin (IL)-6, IL-4, interferon (IFN)-α, and C-reactive protein (CRP) scores was carried out at the beginning and the end of the study. The unpaired variables were examined with independent T-test or the Mann-Whitney test, while the paired variables were examined with paired T-test or Wilcoxon signed rank test. The Spearman correlation test was used to measure correlation between variables. RESULTS: A total of 55 subjects participated and completed the study. The result showed a significant relationship between hydroxychloroquine and decreasing levels of IL-6 and IL-4 (p<0.05). Meanwhile, there was no significant effect on the increase in cognitive function and decrease in IFN-α and CRP (p>0.05) in both groups. CONCLUSION: Hydroxychloroquine decreases the levels of IL-6 and IL-4, but has no effect on cognitive function, levels of IFN-α and CRP.