The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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Viability, Migration Rate, and mRNA Expression of GLUT5, GLUT7, GLUT11 in WiDr Colorectal Cancer Cell Line
Get PDFBACKGROUND: Insufficient glucose levels in colorectal cancer (CRC) patients leads to a condition where fructose might become an alternative source for cells proliferation, but the role of fructose or fructose-glucose combinations in development of CRC has not been elucidated well. In this study, the effect of fructose-glucose variations on viability, migration, and glucose transporter (GLUT)5, GLUT7, GLUT11 mRNA expressions in WiDr CRC cell line were examined.METHODS: Cells were treated with varying ratios of fructose-glucose (F100%; F75%:G25%; F50%:G50%; F25%:G75%; G100%; F: Fructose, G: Glucose). Untreated cells (F0:G0) were used as cell control. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl-2H-tetrazolium bromide (MTT) assay was used for cell viability test, scratch assay was used to examine the cell migration, and quantitative polymerase chain reaction (qPCR) was performed to examine mRNA expressions. Data were analyzed using one-way analysis of variance (ANOVA) and followed with Tukey's post-hoc test, with p<0.05 consideres as significant.RESULTS: Fructose-glucose combinations and glucose 100% significantly increased the cell viability compared to control (p<0.05). All treatment groups showed a significant increase in cell migration compared to control (p=0.000). Only GLUT7 and GLUT11 expressions in the G100% group were significantly different compared to the control (p=0.000). GLUT7 and GLUT11 expressions were also significantly different in F100% and F50%:G50% treatments compared to G100% (p=0.000).CONCLUSION: Taken together, fructose might play important role in cell migration. However, in cell viability, combination with glucose could increase fructose's effect. Fructose might not affect the mRNA expressions of GLUT5, GLUT7 and GLUT11.KEYWORDS: GLUT5, GLUT7, GLUT11, fructose transporter, colorectal cancer, WiD
2-(3-(chloromethyl)benzoyloxy)benzoic Acid Increases CD4+ Regulatory T-Cell Population and FoxP3 Expression in Lipopolysaccharide-induced Mice
Get PDFBACKGROUND: 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.KEYWORDS: 2-(3-(chloromethyl)benzoyloxy)benzoic acid, acetylsalicylic acid, CD4, T-regulatory cells, FoxP3, LP
Ganoderma lucidum Polysaccharide Peptide Reduces Oxidative Stress and Improves Renal Function in Patient with Cardiometabolic Syndrome
Get PDFBACKGROUND: Cardiometabolic syndrome is a risk factor for the development of diseases related to cardiovascular disease and decreased renal function. Ganoderma lucidum polysaccharide peptide (GLPP) has been reported to have anti-inflammatory and antioxidant properties. The current study was conducted to investigate the role of GLPP in inflammatory, oxidative stress and renal function markers of cardiometabolic subjects.METHODS: A randomized double-blinded perspective control trial with pre-post design was conducted. Cardiometabolic syndrome subjects were treated with placebo or GLPP for 60 days. Blood serum was collected from each subject before the first capsule consumption and one day after the last capsule consumption. Serum tumor necrosis factor (TNF)-α, high-sensitivity-C-Reactive Protein (hs-CRP) and malondialdehyde (MDA) levels were measured using enzyme-linked immunosorbent assay, while superoxide dismutase (SOD) level was measured using colorimetric assay. Serum urea and creatinine levels were measured using a clinical analyzer. The Cockroft-Gault formula was used to calculate estimated glomerular filtration rate (eGFR).RESULTS: Compared with the control group, the MDA level was significantly reduced, while the SOD level was significantly increased in the GLPP treatment group. Furthermore, serum urea and creatinine were lowered, while eGFR was increased in the GLPP treatment group.CONCLUSION: Treatment of GLPP for 60 days could be beneficial for lowering oxidative stress and improving renal function of patients with cardiometabolic syndrome.KEYWORDS: Ganoderma lucidum, cardiometabolic syndrome, inflammation, oxidative stress, renal functio
Antibodies to Glutamic Acid Decarboxylase-65 is Associated with Total Daily Dose of Insulin Requirement in Children with Type 1 Diabetes
Get PDFBACKGROUND: 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.KEYWORDS: diabetes mellitus, type 1 diabetes, anti-GAD65, c-peptid
Matrix Metalloproteinase-3 Down Regulation and Cell Migration Inhibition in Human Pterygium Fibroblasts by Mitomycin-C, Curcumin and Fibrin Glue
Get PDFBACKGROUND: Pterygium is an ocular surface disease that often occurs in tropical countries with a high recurrence rate. Matrix metalloproteinase-3 (MMP-3) play a key role in the inflammatory process of pterygium. This study aims to investigate the ability of curcumin and fibrin glue (FG) in suppressing the expression of MMP-3, and whether can be expected as adjuvant therapy to reduce pterygium recurrence.METHODS: Human pterygium fibroblasts (HPF) obtained from primary cultured of pterygium were treated with no treatment, curcumin, mitomycin-C (MMC), and FG. MMP-3 expression was analyzed using immunocytochemistry and the intensity measurement was done using ImageJ software. Cell migration was measured by scratching and stratification of fibroblast culture after cell confluence, and assessed for 48 hours.RESULTS: The expression of MMP-3 were lower in the HPF treated with 100 mol/mL curcumin, 200 mol/mL, and FG (2205.84±86.1 pg/mL, 1002.51±25.22 pg/mL, 1131.55±17.71 pg/mL, respectively) in comparison with untreated HPF (4703.49±108.9 pg/mL). The expression of MMP-3 were significantly different between groups (p<0.001). Cell migration of HPF after scratching with curcumin intervention at 200 mol/mL decrease from 178.67±2.85 (24 hours) to 88.83±1.48 (48 hours). Meanwhile the migration in FG group also decrease from 180.4±2.56 (24 hours) to 72.45±1.25 (48 hours).CONCLUSION: Curcumin and FG able to reduce the expression of MMP-3 and inhibit the migration of HPF cells.KEYWORDS: curcumin, mitomycin C, fibrin glue, human pterygium fibroblast, MMP-
Targeting Metastatic Cancer: Disseminated Tumor Cells and Premetastatic Niches
Get PDFBACKGROUND: Metastases are simply known as cancers spread to another part of the body, and often be responsible for the severity of cancer prognosis. Somehow, the complex mechanisms of metastases are not fully understood yet.CONTENT: The characteristic of cancer is akin to a never-healing wound. Cancer cells are plastic and dynamic as they build their niches and developed into metastases, even when they seem dormant. Therefore, cancer cells can survive the immune system. Recent research has shown the distinct biology of metastasis-initiating cell, which leads to tumor development in distant organs, immune surveillance evasion, and co-option of metastatic micro-environments. Effective cancer therapies must consider the regenerative states of metastatic malignancies and have careful observation of patient phenotypes.SUMMARY: This review aimed to provide an insight on genesis and characteristics of metastases, starting from its seeding and dormancy, until the advance phase. Thus, developing therapy for cancer metastases should not start as it grows, but even as earlier strategies since the primary tumor was detected.KEYWORDS: cancer metastasis, DTC, CTC, CSC, dormancy, pre-metastatic niche, plasticit
Anti-proliferative and Apoptotic Activities of Kasturi Tobacco (Nicotiana tabacum L.) Leaf Resinoid on Cervical Cancer Cell
Get PDFBACKGROUND: Cervical cancer has a high rate of morbidity and mortality in women with cancer. Recent studies have found that tobacco (Nicotiana tabacum L.) is a potential source of anti-cancer agents. Hence, this study was conducted to determine the potential of Kasturi tobacco leaf resinoids as apoptotic agents against cervical cell malignancies, since it has not been fully elucidated before.METHODS: The phytochemical diversity of Kasturi tobacco resinoids was generated by gas chromatography-mass spectrometry (GC-MS) analysis followed by spectral similarity to National Institute of Standards and Technology (NIST) database. Cytotoxicity and proliferative activity of HeLa cells treated with Kasturi tobacco resinoids at various concentrations were evaluated by MTT assay. The expression of Caspase-3, cyclooxygenase-2 (COX-2) and heat shock protein 90 (HSP-90) in HeLa cells was analyzed by immunocytochemistry. Next, the migration ability of HeLa cells was observed by the scratch method.RESULTS: Kasturi tobacco resin contains 4,8,13-cyclotetradecatriene-1,3-diol, 1,5,9-trim with α-2,7,11-cembratriene-4,6-diol (α-CBD) structure in the form of a diterpenoid compound with the chemical formula C20H34O2 and a molecular weight of 306 Da. Kasturi tobacco resinoid with IC50 value of 2500 μg/mL inhibited proliferative activity during 72 hours. At a concentration of 1¼ IC50 and incubation for 48 hours, Caspase-3 expression increased by 74.1%, while COX-2 and HSP-90 expression decreased by 28.3% and 26.1%, respectively. HeLa cell migration was inhibited by Kasturi tobacco resinoid at 24 hours incubation.CONCLUSION: Kasturi tobacco resinoids with a concentration of 1¼ IC50 have potential as cervical anti-cancer agents by increasing Caspase-3 expression and decreasing COX-2 and HSP-90 expression within 48 hours.KEYWORDS: Kasturi tobacco resinoids, cervical cancer, anti-cancer agent, proliferative activity
Vaginal Acidity Affects Vaginal Microbiota in Postmenopausal Women
Get PDFBACKGROUND: The changes in vaginal acidity impact the composition of the vaginal microbiota, either commensal or pathogenic. After menopause, the vaginal tract is more susceptible to infection. Current study was conducted to analyze the effect of vaginal acidity changes on the vaginal microbiota composition in menopausal women.METHODS: A cross-sectional study was conducted on 32 subjects with vulvovaginal atrophy (VVA). Vaginal pH was measured using a strip with colorimetric examination. The detection of Candida sp. was done by using 10% potassium hydroxide. Meanwhile for detection of Trichomonas vaginalis, Gardnerella vaginalis, Lactobacillus iners, and Lactobacillus crispatus, polymerase chain reaction was performed. The data were statistically analyzed.RESULTS: G. vaginalis was the mostly found pathogenic microorganism in current study (40.63%), followed by Candida sp. (25%). Further analysis showed that G. vaginalis were found in L. crispatus positive samples for 9 cases and L. iners positive samples for 9 cases. Candida sp. had an increased risk at vaginal pH ≥6 (OR=8.273), T. vaginalis had a reduced risk at vaginal pH ≥6 (OR=0.765), G. vaginalis had an increased risk at vaginal pH ≥6 (OR=1.440), L. crispatus had an reduced risk at vaginal pH ≥6 (OR=0.077), while L. crispatus had an increased risk at vaginal pH ≥6 (OR=1.111).CONCLUSION: Vaginal acidity alterations in postmenopausal women affect either commensal or pathogenic microorganism composition. A decrease in the number of L. crispatus and an increase in the number of L. iners and pathogenic microorganisms is in line with the increase of pH.KEYWORDS: Lactobacillus, microbiota, menopause, pathogenic microorganisms, vaginal acidit
Curcumin’s Antioxidant Properties in Stable Coronary Artery Disease Patients Undergoing Percutaneous Coronary Intervention: A Randomized Controlled Trial
Get PDFBACKGROUND: Percutaneous coronary intervention (PCI) is the most common intervention for coronary artery disease (CAD) with very low complications. High oxidative stress post-PCI is associated with further atherosclerosis progression. Curcumin, extracted from a specific type of herbs, exhibits anti-oxidant properties by acting as hydrogen and electron donor for superoxide radicals. The aim of this study is to determine the effect of curcumin’s antioxidant properties in reducing oxidative stress of post-PCI in stable CAD.METHODS: This study was a double-blind parallel randomized controlled trial among 50 stable CAD patients undergoing PCI in Cipto Mangunkusumo General Hospital and Jakarta Heart Center. The subjects received either 45 mg/day curcumin or placebo 7 days pre-PCI until 48 hours post-PCI. Reduced oxidative stress markers (decreased MDA or increased GSH) were measured in 3 phases (7 days pre-PCI, 24 hours post-PCI, 48 hours post-PCI).RESULTS: Curcumin group showed increased MDA from baseline to 24 hours (Δ1=0.01 vs. 0.03; p=0.3) and decreased MDA from baseline to 48 hours (Δ2=-0.06 vs. 0.03; p=0.9). While, curcumin group showed decreased GSH from baseline to 24 hours (Δ1=-49.7% vs. 12.2%; p=0.4) and from baseline to 48 hours (Δ2=-19.09% vs. 11.4%; p=0.6). However, no significant changes were found in malondialdehide (MDA) and glutathione (GSH) level after the intervention.CONCLUSION: The 45 mg/day curcumin supplementation from 7 days pre-PCI until 48 hours post-PCI had no significant antioxidant effect in stable CAD post-PCI.KEYWORDS: coronary artery disease, curcumin, antioxidant, percutaneous coronary interventio
High NF-κB and RAGE Expression in Fetal Membrane of Premature Rupture of Membrane (PROM) Subject
Get PDFBACKGROUND: Premature Rupture of Membranes (PROM) is significantly linked to the infections-related maternal deaths. In the inflammatory process, the influencing stressor will stimulate the activation of Nuclear Factor Kappa B (NF-κB) and Receptor of Advanced Gyclation End product (RAGE). Yet up to date, the expression of NF-κB and RAGE in pregnant women with PROM are still rarely studied. Therefore, this study aimed to observe the differences of NF-κB and RAGE expression from PROM and non-PROM subjects.METHODS: This was a cross-sectional study involving 20 PROM subjects and 20 non-PROM subjects with infections and complications. Samples from the fetal membrane tissue of subjects were obtained and put into paraffin block preparation for the determination of NF-κB and RAGE expression. The detection of NF-κB and RAGE expression was conducted using immunohistochemical staining and observed under an upright light microscope. The expressions were later calculated using ImageJ software. RESULTS: Both NF-κB and RAGE expression were found to be higher in PROM subjects compare to the non-PROM subjects. The median of NF-κB in PROM and non-PROM subjects were 32.47±1.22 and 5.59±1.09, respectively (p=0.000). While the median of RAGE in PROM subjects was 53.58±3.46, and in non-PROM subjects was 11.64±2.49 (p=0.013).CONCLUSION: There is significant difference between NF-κB and RAGE expression in fetal membranes of PROM and non-PROM subjects. Therefore, the increased of NF-κB and RAGE expression can be used as a potential marker to detect complication of PROM.KEYWORDS: premature ruptures of membrane, non-premature ruptures of membrane, expression of NF-κB