The Indonesian Biomedical Journal (Prodia Education and Research Institute)
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Thymoquinone Modulates Local MMP-9, IL-10, and IgG in Sciatic Nerve Crush Injury Animal Model
Get PDFBACKGROUND: Interleukin (IL)-10 is involved in Wallerian degeneration after peripheral nerve crush injury. Oral thymoquinone was previously observed to decrease local immunoglobulin-G (IgG) in a crush-injured rat model. No study has evaluated the pathway of various thymoquinone dosages on local IgG and IL-10 in this injury.METHODS: This experimental study used 126 Rattus norvegicus Wistar rats that were divided into 18 groups: six groups received a placebo, the other six groups received thymoquinone at 100 mg/kg/day and the last six groups received thymoquinone at 250 mg/kg/day, respectively. Rats were sacrificed at 12, 18, 24, 5x24, 6x24, and 7x24 hours. Matrix metalloproteinase-9 (MMP-9), IL-10, and local IgG levels were assessed by Enzyme-Linked Immunosorbent Assay (ELISA). The nuclear factor KappaB (NF-κB) expressions on Schwann cells were examined by flow cytometry. Path analysis was performed using SmartPLS.RESULTS: The path analysis showed that 100mg/kg/day of thymoquinone significantly decreased NF-κB expression. However, NF-κB did not affect local MMP-9, and MMP-9 had no significant relationship with local IL-10 and IgG. Thymoquinone 250 mg/kg/day also significantly inhibited NF-kB expression, decreased local MMP-9, and, in turn, decreased local IL-10 and IgG.CONCLUSION: Administration of oral thymoquinone 250 mg/kg/day decreases local IgG and IL-10 levels via suppressing NF-κB expression and MMP-9 levels.KEYWORDS: thymoquinone, crush injury, IgG, IL-10, MMP-9, NF-κ
Caffeic Acid Inhibits Tumour Mass Formation in MG-63 Cells-induced Nude Mice
Get PDFBACKGROUND: Formation of tumour mass is one symptom of osteosarcoma development. Caffeic acid has been known to provide effective treatment but has less side effect for some cancer therapy. Studies reported that caffeic acid might promote apoptosis in MG-63 osteosarcoma cells, however, the effect of caffeic acid treatment in preventing tumour mass formation has not been well elucidated, especially in MG-63 cells-induced nude mice in vivo.METHODS: MG-63 cells were pre-treated with 0, 1, or 10 µg/mL caffeic acid, and 6 hours after pre-treatment, MG-63 cells were injected into subcutaneous space of mice to induce osteosarcoma. Another model was also created by subcutaneously injecting MG-63 cells to the back of mice, and after 48 days, the visible tumour mass was injected intra-tumour with 0 or 10 µg/mL caffeic acid every 7 days for 6 times. After 90 days, mice were anaesthetised, and the nodule pictures were taken for observation and measurement. RESULTS: In pre-treated MG-63 cells-induced mice, volumes of the mass decreased in reverse with the dose of caffeic acid given. Ten µg/mL caffeic acid pre-treatment was able to significantly lower the mass volume compared to the untreated (p<0.05). Meanwhile, the intra-tumour treatment of 10 µg/mL caffeic acid, even though not significant, was able to inhibit tumour mass formation.CONCLUSION: Results of caffeic acid pre-treatment and caffeic acid treatment in tumour mass of mice show that caffeic acid is able to inhibit the MG-63 cells formation. This suggests that caffeic acid can be a potential anti-cancer agent.KEYWORDS: caffeic acid, osteosarcoma, MG-63 cells, tumour mas
Parkia speciosa Seeds Ethanol Extract as Co-chemotherapeutic Agent for Doxorubicin Toward Tongue Cancer
Get PDFBACKGROUND: Parkia speciosa seeds have been reported to have an anticancer property due to the presence of various antioxidant compounds. Since the potential uses of P. speciosa for the tongue cancer has not been clearly disclosed, we conducted a study to investigate anticancer properties of P. speciosa seed ethanol extract (PSSEE) as well as its effect on cardiac cells.METHODS: Tongue cancer rat model were treated with/without Doxorubicin and various concentrations of PSSEE. After treatment, tongue and heart samples were collected and processed further for histological examinations. Tongue epithelium thickness and damaged heart tissues was observed by HE staining, while tongue cancer cell proliferation was assessed by Ki-67 immunohistochemistry. Analyses were performed under an upright light microscope to measure tongue epithelium thickness, state of cancer cell proliferation, and degree of heart tissue damage.RESULTS: Addition of 400 mg/kg body weight (BW) PSSEE to 4.6 mg/kg BW Doxorubicin reduced the average tongue epithelial thickness and Ki-67+ cells number. Upon addition of PSSEE to Doxorubicin, the damage of heart tissue was reduced in a concentration dependent manner. Among all groups, the group of tongue cancer treated with 4.6 mg/kg BW Doxorubicin and 400 mg/kg BW PSSEE had the lowest percentage as well as the lowest degree of heart tissue damage.CONCLUSION: Since addition of PSSEE to Doxorubicin reduced epithelial thickness, number of Ki-67+ cells and heart tissue damage, PSSEE could be a potential co-chemotherapeutic agent for Doxorubicin toward tongue cancer.KEYWORDS: Parkia speciosa, Doxorubicin, tongue cancer, epithelial thickness, Ki-67, cardiotoxicity, co-chemotherap
Analysis of Single Nucleotide Polymorphisms on Locus 13q33.1-34 in Multigenerational Families of Cleft Lip Palate using MassArray
Get PDFBACKGROUND: Cleft lip palate is a common congenital anomaly with multifactorial etiology. Many high-risk markers at different loci were reported to be involved in its etiology. Advanced genetic research led to the discovery of evidence of a new linkage on 13q33.1-34 region at marker rs1830756 in two multigenerational Indian families. However, no further study was reported to confirm or validate this linkage in other families. Hence, the present study was designed.METHODS: Twenty multigenerational families affected by non-syndromic cleft lip palate were selected for the study. Polymorphisms, rs1830756, rs1323672, rs1935135 of FAM155A gene; rs1961495, rs953386, rs1411040 of COL4A1 gene; and rs726449, rs984300 of MYO16 gene were selected. Genomic DNA was isolated and sent for genetic analysis by single nucleotide polymorphism (SNP) genotyping using the MassArray method. Statistical analysis of the genomic data was done by PLINK. Bonferroni correction was applied and haplotype analysis was done using Haploview software.RESULTS: Polymorphisms followed the Hardy Weinberg Equilibrium. In the allelic association, all the polymorphisms analysed showed no statistical significance. Hence, there was no significant difference in the allelic frequencies between non-syndromic cleft lip palate patients and healthy controls. The odds ratio was not more than 1.6 for all the SNPs. Haplotype analysis showed that haplotypes were not significantly higher in non-syndromic cleft patients than in control subjects.CONCLUSION: There is no association between SNPs analysed in the locus 13q33.1-34 with cleft lip palate.KEYWORDS: cleft lip palate, chromosome, polymorphis
Suppression of MiR130a-3p Using CRISPR/Cas9 Induces Proliferation and Migration of Non-Small Cell Lung Cancer Cell Line
Get PDFBACKGROUND: Molecular alterations of microRNA130a (miR130a) are observed in many types of cancers, including non-small cell lung cancer (NSCLC). However, the role of miR130a in NSCLC has been poorly studied.METHODS: In this study, clustered regularly interspaced short palindromic repeat (CRISPR)-Cas9 was utilised to knockdown miR130a. The gRNA was designed to target the stem loop, 3’ and 5’ sites of miR130a and stably expressed in A549 cells. Post-treatment, mature levels of miR130a-3p and 5p were quantified, and proliferation and migration assays were conducted.RESULTS: Result showed significant suppression of miR130a-3p and -5p by two and three-fold respectively, when the CRISPR/Cas9 targeted at the 3’ site and stem loop of the miR130a gene. Suppression of miR130a-3p significantly increased the growth and migration of A549 cells, but no significant changes were observed in cells with suppressed expression of miR130a-5p.CONCLUSION: Our encouraging results highlight that the suppression of the miR130a is achievable using CRISPR/Cas9, and suppression of the miR-130a-3p could play an important role in the regulation of NSCLC.KEYWORDS: miR130a, CRISPR-Cas9, non-small cell lung cance
Association of 25-hydroxyvitamin D, Cyclooxygenase-2 and Prostaglandin E2 Serum Levels in Breast Cancer Patients
Get PDFBACKGROUND: Serum levels of 25-hydroxyvitamin D (25(OH)D), prostaglandin E2 (PGE2), and cyclooxygenase 2 (COX2) expression differ between breast cancer stages. Since, previous studies showed mixed results, in this study, we aimed to analyze vitamin D levels related to breast cancer stages and serum levels of COX2 and PGE2 in Indonesia.METHODS: This was a cross sectional study involving 75 breast cancer patients. Subjects were divided into 3 groups, namely operable early stage (K1), locally advanced stage (K2), and advanced stage (K3). Venous blood samples were taken from each subject, then were analyzed for the 25(OH)D, COX2, and PGE2 serum levels by enzyme-linked immunosorbent assay (ELISA) method.RESULTS: There were significant differences in 25(OH)D among groups (p=0.012); between K1 and K2 (p=0.009) and between K1 and K3 (p=0.023). However, there was no significant difference in serum COX2 level (p=0.328). There were significant differences of PGE2 among groups (p=0.002); between K1 and K2 (p=0.036) and between K1 and K3 (p=0.001). Correlation test showed that there were differences between 25(OH)D serum levels and PGE2 serum level (r=0.306, p=0.008) and also between 25(OH)D serum level and breast cancer stage (r=-0.229; p=0.048).CONCLUSION: There were differences in serum Vitamin D and PGE2 levels at various stages of breast cancer. Serum 25(OH)D levels had weak correlation with breast cancer stage and PGE2 serum level. Serum vitamin D level in advanced breast cancer were lower than early stage breast cancer and indicate a poor prognosis.KEYWORDS: breast cancer, 25-hydroxyvitamin D, cyclooxygenase 2, prostaglandin E
Comparison of Light Transmission Aggregometry and VerifyNow in Detecting Clopidogrel Resistance and Factors Affecting Clopidogrel Resistance in AMI-EST Patients Undergoing Percutaneous Coronary Intervention: A Cross-Sectional Study
Get PDFBACKGROUND: Light transmission aggregometry (LTA) and VerifyNow is commonly used to measure platelet responsiveness to clopidogrel. This study aimed to compare the results of LTA and VerifyNow P2Y12 assay for assessing the clopidogrel resistance in patients undergoing percutaneous coronary intervention and determine factors affecting clopidogrel resistance.METHODS: The subjects were 119 patients who underwent percutaneous coronary intervention (PCI) and had given loading dose of 600 mg clopidogrel. Blood samples were taken at 6 hour after clopidogrel loading dose. Platelet aggregation was measured by LTA and VerifyNow.RESULTS: LTA and VerifyNow assay showed fair agreement with Kappa=0.270, p=0.001. The proportion of resistance to clopidogrel using VerifyNow was 21.8% and LTA was 47.1%. Patients with diabetes melitus were more likely to develop clopidogrel resistance than patients without diabetes (OR of 7.67; 95% CI: 1.87-31.50; p=0.005).CONCLUSION: The ability of LTA and VerifyNow in detecting clopidogrel resistance were not comparable. Multivariate analysis results for VerifyNow shows diabetes mellitus as the greatest predictors of clopidogrel resistance.KEYWORDS: agreement, clopidogrel resistance, LTA, predictor, VerifyNow
Metabolic Reprogramming and Molecular Rewiring in Cancer: Therapeutic Opportunities
Get PDFBACKGROUND: A lot of contemporary cancer research has concentrated on genetic influence. However, cancer also involves biochemical changes, such as metabolic adaptation to support the aberrant cell proliferation.CONTENT: The fast cell proliferation in cancer cells enforce a metabolic re-arrangement to promote their long-term survival. The increased glucose uptake and fermentation of glucose to lactate are common features of this altered metabolism known as “the Warburg effect”. These metabolic pathways regulation enable cancer cells to produce adenosine triphosphate (ATP) in an efficient way. Epigenetic and metabolic changes also both affect molecular rewiring in cancer cells and promote cancer development and progression.SUMMARY: Metabolic rewiring and epigenetic remodeling establishing a direct link between metabolism and nuclear transcription to promote the survival of tumor cells. A further understanding of how metabolic remodeling can result in epigenetic changes in tumors, affecting cancer cell differentiation, proliferation, and/or apoptosis, will lead to a new strategy for cancer therapy.KEYWORDS: cancer metabolism, epigenetics, metabolic reprogramming, molecular rewirin
The Effects of Propolis Extract Administration on HIV Patients Receiving ARV
Get PDFBACKGROUND: Human Immunodeficiency Virus (HIV) is an infectious disease that targets the human immune system by attacking cluster of differentiation (CD)4 cells. The use of propolis in HIV patients is expected to be safe and beneficial in terms of increasing endurance and immunity by its role in increasing CD4 level. This study aimed to analyze the influence of propolis supplementation in increasing the CD4 level in anti-retroviral (ARV)-treated HIV patients.METHODS: Double-blind randomized controlled clinical trial was conducted in 50 HIV patients who took regular ARV therapy. The subjects were divided into two groups, one group was treated with ARV and propolis, while another one was given ARV and placebo. The CD4 cell count was measured during pre-treatment, in the 3rd month, in the 6th month after treatment. The level of hemoglobin, leukocyte, and platelets were also measured. The SF-12 questionnaire was used to evaluate quality of life of the subject.RESULTS: Out of 50 subjects, 43 subjects completed the study, which were 19 subjects from the propolis group and 24 subjects from the placebo group. After 3-month of treatment, there was a statistically significant difference in the increase of CD 44 level in propolis group, while the increment was not significant in the placebo group. After 6-month treatment, the increase of CD4 level was occurred in both groups, propolis and placebo, however the increment was not statistically significant. The levels of hemoglobin, leukocyte, and platelets were not altered by the treatment and remained normal throughout the study. The quality of life was improved during the study; however, it was also not statistically significant. Mild adverse events occurred in 3 subjects which were relieved after the treatment stopped.CONCLUSION: Based on the result of this study, the administration of propolis on HIV patients receiving ARV bring significant difference in the increase of CD4 in propolis group from baseline to 3 month after the treatment. While in placebo group, this increment was not significant. At the end of study, CD4 count continued to rise up, however the increase was not statistically significant. There are no hemoglobin, leukocyte, platelets, and quality of life abnormalities. Therefore, it is necesary to do further research with a spesific CD4 count. However, it may be beneficial in relieving the clinical symptoms and quality of life of patient living with HIV.KEYWORDS: CD4, ARV, HIV, propoli
Robusta Extract Cream Ameliorated Ultraviolet B-induced Wrinkle Skin of Mice by the Regulation of Epidermal Thickness and Inhibition of MMP-1
Get PDFBACKGROUND: Recently, coffee is widely used for preventing photoaging because of its antioxidant capacity. Among two kinds of coffee, robusta coffee has higher content of antioxidant such as chlorogenic acid (CGA) and caffeine. Researchs about robusta coffee bean effect on photoaging due to UVB radiation is still limited. Therefore, the aim of this study was to examine the effect of robusta extract cream (RE cream) on preventing wrinkle in mice induced by ultraviolet-B (UVB) radiation.METHODS: RE cream was made by mixing RE coffee with moisturizing cream in different concentration (10%, 20%, and 40%). Twenty-five male of Mus musculus Balb/c strain mice aged 4 weeks were divided into five groups; control group, UVB group, UVB + 10% RE group, UVB + 20% RE group, and UVB + 40% RE group. The UVB groups were given UVB radiation three times a week with an exposure duration of 100 seconds per time for ten weeks. At the end of the treatment, skin samples were excised and statined histologically, also were analyzed for their protein expression. Evaluation of wrinkles was carried out using the Bissete method before and after treatment. To evaluate the thickness of the epidermis, HE staining was performed, while masson Trichome staining was performed to determine the collagen content.RESULTS: RE cream-treated groups showed lower wrinkle score compared to the control group. Furthermore, in UVB + 10% RE group, the RE cream application reduce wrinkle formation. In UVB + 10% RE group and UVB + 20% RE group, the RE cream application increased epidermal thickness and collagen content (p=0.00). While collagenase, matrix metalloproteinase-1 (MMP-1) expression was lower in UVB + 20% RE group compared to the UVB group (p<0.05), however the MMP1 expression in UVB + 40% RE group was higher than other treatment group.CONCLUSION: RE cream prevents wrinkle by maintaining epidermal thickness and collagen contain. RE cream also decreases MMP-1 expression in mice.KEYWORDS: coffee, collagen, MMP-1, robusta, wrinkl