Nur Permatasari

Vasa vasorum anti-angiogenesis through H2O2, HIF-1α, NF-κB, and iNOS inhibition by mangosteen pericarp ethanolic extract (Garcinia mangostana Linn) in hypercholesterol-diet-given Rattus norvegicus Wistar strain

Background Oxidative stress in atherosclerosis produces H2O2 and triggers the activation of nuclear factor kappa beta (NF-κB) and increase of inducible nitric oxide synthase (iNOS). The formation of vasa vasorum occurs in atherosclerosis. Vasa vasorum angiogenesis is mediated by VEGFR-1 and upregulated by hypoxia-inducible factor-1α (HIF-1α). The newly formed vasa vasorum are fragile and immature and thus increase plaque instability. It is necessary to control vasa vasorum angiogenesis by using mangosteen pericarp antioxidant. This study aims to demonstrate that mangosteen pericarp ethanolic extract can act as vasa vasorum anti-angiogenesis through H2O2, HIF-1α, NF-κB, and iNOS inhibition in rats given a hypercholesterol diet. Methods This was a true experimental laboratory, in vivo posttest with control group design, with 20 Rattus norvegicus Wistar strain rats divided into five groups (normal group, hypercholesterol group, and hypercholesterol groups with certain doses of mangosteen pericarp ethanolic extract: 200, 400, and 800 mg/kg body weight). The parameters of this study were H2O2 measured by using colorimetric analysis, as well as NF-κB, iNOS, and HIF-1α, which were measured by using immunofluorescence double staining and observed with a confocal laser scanning microscope in aortic smooth muscle cell. The angiogenesis of vasa vasorum was quantified from VEGFR-1 level in aortic tissue and confirmed with hematoxylin and eosin staining. Results Analysis of variance test and Pearson’s correlation coefficient showed mangosteen pericarp ethanolic extract had a significant effect (P<0.05) in decreasing vasa vasorum angiogenesis through H2O2, HIF-1α, NF-κB, and iNOS inhibition in hypercholesterol-diet-given R. norvegicus Wistar strain. Conclusion Mangosteen pericarp ethanolic extract 800 mg/kg body weight is proven to decrease vasa vasorum angiogenesis. Similar studies with other inflammatory parameters are encouraged to clarify the mechanism of vasa vasorum angiogenesis inhibition by mangosteen pericarp ethanolic extract.

Combined effects of shear stress and glucose on the morphology, actin filaments, and VE-cadherin of endothelial cells in vitro

Objective The purpose of the present study was to analyze the effects of glucose and shear stress on the morphology and density of endothelial cells, actin filamen, and the expression of VE-cadherin. Methods After confluency of endothelial cells (3–4 days), 22 mM of d-glucose was administered for 7 days. Endothelial cells were exposed to shear stress of 10 dyne/cm2 for varied durations of 5, 8, 12, and 15 min. Morphology of ECs was observed using an inverted microscope before and after shear stress exposure. VE-cadherin and actin filament were analyzed immunohistochemically. Results Exposure to high glucose induces more shrinkage and the cell density decreased at 15 min. High glucose reduced actin filaments and the more globular ones, especially around the nuclei. There was a decline in VE-cadherin scores with significant differences between treatments with 5 mM and 22 mM of glucose. Conclusion Combination exposure of shear stress and high glucose changes morphology, reduces actin filament and VE-cadherin, of endothelial cells.

Evaluation Anxiolytic Effect of Methanol Extract of Ceplukan Leaves (Physalis minima L.) in the Elevated Plus Maze Test through IL-6 Level Changes in Ovariectomized Rats

Ceplukan ( Physalis minima L.) has long been used to treat various conditions in traditional medicine. This study aims to demonstrate the anxiolytic effects of Methanol Extract of Ceplukan Leaves (MECL) in the Elevated Plus Maze (EPM) test and correlate to IL-6 level of ovariectomized rat brain. Total of 24 Wistar rats were divided into six groups: one normal group, one group of 1 month ovariectomized (ovx), one group of 2 months ovx, three groups of 2 months ovx (each given with MECL 500; 1500 and 2500 mg/kg doses for 1 month). The anxiety-like behavior level was measured by EPM test. After EPM test, the brain was removed to measure level of IL-6 by ELISA. The data were processed and analyzed by one-way ANOVA and Pearson correlation. We found that the MECL-treated rats enter the opened-arm higher than the control rats. It indicates that the MECL-treated rats are less anxious than the control rats. The results also show the decreased of IL-6 level in MECL-treated rats.

EFFECT OF METHYLGLYOXAL ON SOLUBLE RECEPTOR FOR ADVANCED GLYCATION END PRODUCTS (sRAGE) OF PRE-OSTEOBLAST MC3T3E1 CELL LINE -

Methylgyoxal (MG) is reactive dicarbonyl compound was found in high level in blood of diabetic patients. Methlyglyoxal was more reactive compared to glucose and have toxic effect through AGEs. Soluble Receptor for Advanced Glycation End Products (sRAGE) act as a scavenger and decoy receptor for AGEs. This study aimed to investigate the effect of changes intracellularly redox status on sRAGE levels in preosteoblast MC3T3E1 exposed to MG. The changes of redox status was obtained by blockade of superoxide dismutase (Diethylthiocarbamoic acid/DETCA), blockade of gluthatione peroxidase (mercapto succinate/MS), and chelating of iron (defferoxamine/DFX). sRAGE levels was evaluated by enzyme linked imunnosorbent assay technique. The levels of sRAGE in MG-exposed group or DETCA-exposed group was significantly increased than that in control group (P < 0.05). The levels of sRAGE in MG+DETCA-exposed group was significantly decrease compared with DETCA-exposed group (P < 0.05). The levels of sRAGE was significantly increased in MS-exposed group or MG+MS-exposed group than that in control group, respectively (P < 0.05). Concluded that the change of redox status by antioxidant blocker or MG exposure would increases sRAGE levels in preosteoblast MC3T3E1 cell line. The modulation of redox state by antioxidant blocker does not affect sRAGE level in the presence of MG. Iron chelating does not change the sRAGE level.

Effect of methylglyoxal on osteoprotegerin expression in the pre-osteoblast MC3T3E1 cell line: involvement of oxidant pathways

Objective: This study aimed to evaluate osteoprotegerin (OPG) expression in MC3T3E1 preosteoblast cell line exposed to methylgyoxal (MG) with/without antioxidant blocking or iron chelating. Methods: MC3T3E1 preosteoblast cell line subclone 4 was obtained from American Type Culture Cell Collection (ATCC). Modulation of intracellular oxidative status was obtained by blockade of superoxide dismutase (SOD) using diethylthiocarbamoic acid (DETCA), blockade of gluthatione peroxidase (GPx) using mercaptosuccinate (MS), and iron chelating using deferoxamine (DFX). The MC3T3E1 preosteoblast cell line was exposed to MG at the concentration of 5 µM for 6 h of incubation time. Eight groups (n=3) were constituted, including the control group, the MG-exposed group, the DETCA/MS/DFX-exposed groups, and the MG+DETCA/MS/DFX-exposed groups. OPG was evaluated by ELISA assay kit. Results: The expression of OPG in MG-exposed group was not different than that in the control group. OPG expression was significantly reduced by DETCA and restored in the MG+DETCA-exposed group. MS exposure also significantly reduced OPG expression; addition of MG to MS again clearly reversed this effect. On the other hand, DFX administration did not affect OPG expression. Conclusion: Modulation of the intracelllular oxidative status via blockade of SOD or GPx clearly inhibits the OPG expression in preosteoblast cells. Addition of MG reversed this effect and reversed the fall of OPG expression.