Claim Missing Document
Check
Articles

Found 3 Documents
Search
Journal : The Indonesian Biomedical Journal

Negative Correlation between Cytoglobin Expression and Intracellular ROS Levels in Human Skin Keloid Fibroblasts Fajri Marindra Siregar; Novi Silvia Hardiany; Sri Widia Azraki Jusman
The Indonesian Biomedical Journal Vol 11, No 1 (2019)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v11i1.488

Abstract

BACKGROUND: In our previous study, we found higher cytoglobin (Cygb) expression in keloid than normal tissue. Cytoglobin is a new globin family protein which function is still being studied to date. The purpose of this research is to elucidate the function of Cygb in human skin keloid fibroblasts (KFs), especially its role in intracellular reactive oxygen species (ROS) levels.METHODS: The study was conducted on human skin KFs obtained from primary culture. Inhibition of Cygb expression was achieved by using siRNA targeting Cygb. We compared the relative expression of Cygb between treatment and control group, and its effect on intracellular ROS levels. Gene expression was measured using quantitative real-time polymerase chain reaction (qRT-PCR) while the ROS level counted by dichlorodihydrofluorescein diacetate (DCFHDA) assay.RESULTS: There was an increase in intracellular ROS levels in the small interfering RNA (siRNA) (+) Cygb group compared to control group (1.673 vs. 1.260; 1.773 vs. 1.393; 1.710 vs. 1.360; respectively). There is a negative correlation between Cygb expression and ROS level (p<0.05; r=-0.651).CONCLUSION: There is a negative correlation between Cygb expression and intracellular ROS levels, we suggest Cygb acts as a ROS scavenger in human skin KFs.KEYWORDS: skin keloid fibroblasts, cytoglobin, siRNA, ROS
TNFR, TRAF2, NF-κB mRNA Levels of Glioblastoma Multiforme Cells Treated by Conditioned Medium of Umbilical Cord-derived Mesenchymal Stem Cells Novi Silvia Hardiany; Yohana Yohana; Septelia Inawati Wanandi
The Indonesian Biomedical Journal Vol 11, No 2 (2019)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v11i2.722

Abstract

BACKGROUND: Glioblastoma multiforme (GBM) is a human malignant brain tumor which is arise from glial cells. Our previous study proved that GBM cells proliferation increased after treating by conditioned medium of umbilical cord-derived mesenchymal stem cells (CM-UCSCs). Cells proliferation is probably mediated by tumor necrosis factor (TNF)-α which could bind to membrane receptor and induce signaling pathway. Therefore, this research was intended to analyze the mRNA expression of TNF-α signaling pathway molecules on CM-treated GBM cells by measuring TNF receptor 1 and 2 (TNFR1 and TNFR2), TNFR associated factor 2 (TRAF2), nuclear factor kappa B (NF-κB) mRNA level, and TNFR2 protein level.METHODS: UCSCs and human glioblastoma T98G cells were cultured and harvested after 80% confluence. CM was prepared by growing UCSCs in serum alpha Minimum Essential Media (α-MEM) for 24 hours. Fifty percent concentration of CM-UCSCs was used to treat T98G cells for 24 hours. TNF-α level in CM-UCSC was detected using enzyme linked-immunosorbent assay (ELISA), while the expression of TNFR1, TNFR2, TRAF2 and NF-κB were detected using quantitative Reverse Transcriptase Polymerase Chain Reaction (qRT-PCR), and TNFR2 protein level was detected using sandwich ELISA.RESULTS: TNF-α level was detected in CM-UCSCs 4.4 pg/mL. Moreover, the expression of TNFR1, TNFR2, TRAF2 and NF-κB were significantly 1.4-fold, 4.9-fold, 5.6-fold, 1.8-fold respectively higher in T98G treated cells than control. TNFR2 protein level in T98G treated cells was 11.57 pg/mg protein higher than control.CONCLUSION: The expression of molecules involved in TNF-α signaling pathway were up regulated in T98G cells treated by CM-UCSCs.KEYWORDS: CM-UCSCs, TNFR1, TNFR2, TRAF2, NF-κB, T98G cells
Andrographis paniculata Ethanol Extract Alleviates High Glucose-induced Senescence of Human Umbilical Vein Endothelial Cells via the Regulation of mTOR and SIRT1 Pathways Khatimah, Nurul Gusti; Arozal, Wawaimuli; Barinda, Agian Jeffilano; Antarianto, Radiana Dhewayani; Hardiany, Novi Silvia; Shimizu, Ippei; Fadhillah, Muhamad Rizqy
The Indonesian Biomedical Journal Vol 16, No 4 (2024)
Publisher : The Prodia Education and Research Institute (PERI)

Show Abstract | Download Original | Original Source | Check in Google Scholar | DOI: 10.18585/inabj.v16i4.3067

Abstract

BACKGROUND: Chronic exposure of high glucose (HG) in endothelial cell induces senescence which may contribute to the development and progression of age-related diseases including insulin resistance. Andrographis paniculata improves insulin resistance in recent in vitro and in vivo studies. Anti-inflammatory and antioxidant properties of A. paniculata may be the new therapeutic approach to inhibiting premature senescence. However, the senolytic effect of A. paniculata on endothelial cells has not been investigated comprehensively. This study was conducted to evaluate the effect of A. paniculata extract on HG-induced endothelial cell senescence and the underlying mechanisms.METHODS: Human umbilical vein endothelial cells (HUVECs) were treated with 33 mM HG and 7.5 μg/mL A. paniculata extract for 48 hours. The expressions of p16, p21, interleukin (IL)-6, IL-8, insulin receptor substrate (IRS)-1, mammalian target of rapamycin, and sirtuin 1 (SIRT1) were measured by performing real-time quantitative polymerase chain reaction (RT-qPCR). The senescence-associated-β-galactosidase (SA-β-gal) staining was performed to observe the positive-stained senescent cells, while the cell surface expression of IL-1α was examined with flow cytometry method.RESULTS: A. paniculata extract reversed senescence in HUVECs under HG conditions by reducing mRNA expressions of p16 and p21, the number of SA-β-gal-positive-stained cells, and the expression of IL-1α on cell surface, which decreased the activation of IL-6 and IL-8. In addition, A. paniculata extract decreased the mRNA expression of mTOR and increased the mRNA expressions of IRS-1 as well as SIRT1.CONCLUSION: A. paniculata extract ameliorated senescence and improved insulin sensitivity by regulating the mTOR, SIRT1, and IRS-1 mRNA expressions on HG-treated HUVECs.KEYWORDS: Andrographis paniculata, endothelial cell, senescence, high glucose, nutrient-sensing pathways