Effect of steroids on transcription and secretion of Gal-1 by the human trophoblast cell line in vitro

Galectin-1 (Gal-1) is a lectin with recently documented pro-invasive function in trophoblasts in vitro, whose regulation is currently insufficiently known. The potential involvement of steroid hormones, synthetic glucocorticoid dexamethasone (DEX), the sex steroid progesterone (PRG) and mifepristone (RU486) in the regulation of Gal-1 in the trophoblast-derived cell line HTR-8/SVneo was investigated. Gal-1 mRNA levels were assessed by real-time PCR. The effect on secretion of Gal-1 into the culture media was followed using the SELDI-TOF protein chip array. We present evidence that DEX and RU486 significantly reduced Gal-1 in the HTR-8/SVneo cell line at the mRNA level. In addition, trophoblast-derived HTR-8/SVneo cells were shown to secrete detectable Gal-1 protein, which was only slightly increased by PRG. The potential clinical relevance of these findings remains to be determined. [Projekat Ministarstva nauke Republike Srbije, br. 173004

Effect of steroids on transcription and secretion of Gal-1 by the human trophoblast cell line in vitro

Galectin-1 (Gal-1) is a lectin with recently documented pro-invasive function in trophoblasts in vitro, whose regulation is currently insufficiently known. The potential involvement of steroid hormones, synthetic glucocorticoid dexamethasone (DEX), the sex steroid progesterone (PRG) and mifepristone (RU486) in the regulation of Gal-1 in the trophoblast-derived cell line HTR-8/SVneo was investigated. Gal-1 mRNA levels were assessed by real-time PCR. The effect on secretion of Gal-1 into the culture media was followed using the SELDI-TOF protein chip array. We present evidence that DEX and RU486 significantly reduced Gal-1 in the HTR-8/SVneo cell line at the mRNA level. In addition, trophoblast-derived HTR-8/SVneo cells were shown to secrete detectable Gal-1 protein, which was only slightly increased by PRG. The potential clinical relevance of these findings remains to be determined

Supplementary data for article: Stanković, N.; Šenerović, L.; Bojic-Trbojevic, Z.; Vuckovic, I.; Vicovac, L.; Vasiljevic, B.; Nikodinović-Runić, J. Didehydroroflamycoin Pentaene Macrolide Family from Streptomyces Durmitorensis MS405(T): Production Optimization and Antimicrobial Activity. Journal of Applied Microbiology 2013, 115 (6), 1297–1306. https://doi.org/10.1111/jam.12326

Supporting information for: [https://doi.org/10.1111/jam.12326]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1438]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3486

Supplementary data for article: Stanković, N.; Šenerović, L.; Bojic-Trbojevic, Z.; Vuckovic, I.; Vicovac, L.; Vasiljevic, B.; Nikodinović-Runić, J. Didehydroroflamycoin Pentaene Macrolide Family from Streptomyces Durmitorensis MS405(T): Production Optimization and Antimicrobial Activity. Journal of Applied Microbiology 2013, 115 (6), 1297–1306. https://doi.org/10.1111/jam.12326

Supporting information for: [https://doi.org/10.1111/jam.12326]Related to published version: [http://cherry.chem.bg.ac.rs/handle/123456789/1438]Related to accepted version: [http://cherry.chem.bg.ac.rs/handle/123456789/3486

Reticulocyte response in paired blood samples of Babesia canis infected young and adult dogs

Acute B. canis infection can lead to an acute phase reaction (APR) in dogs. The parasite invades red blood cells causing anemia through immune-mediated hemolysis and possible erythropoietic suppression. A regenerative response of the erythroid lineage during the babesiosis has not been described in extension. This research examines hematologic parameters focusing on the absolute reticulocyte count and apolipoprotein A I (ApoA I) level on the day of admission and 14 days after treatment with imidocarb-dipropionate in young (n=11) and adult (n=11) dogs naturally infected with B. canis. Metabolic and inflammatory processes were characterized by analyzing protein and lipid profiles, as well as ApoA I at specified time points. Automated analyzers were used to determine complete blood count and biochemical parameters, while ApoA I was assessed using radioimmunoassay. The reticulocyte count was determined using a manual method by means of supravital staining. Both young and adult dogs with acute B. canis infection showed non-regenerative anemia without difference. Fourteen days after successful treatment with imidocarb-dipropionate, the anemia was corrected and a high reticulocyte count was observed (p<0.05). This indicates that the erythroid regenerative response was efficient in young and adult dogs, although vital signs, leukocyte count and triglyceride concentration suggest a more intense APR in young dogs. A decrease in ApoA I in both groups 14 days after treatment (p<0.01) confirmed that this lipoprotein acts as a positive acute-phase protein in acute B. canis infection in dogs, but further studies are needed to connect its role in erythroid lineage regeneration

Oleuropein Attenuates Oxidative Stress in Human Trophoblast Cells

Olive-derived bioactive compound oleuropein was evaluated against damage induced by hydrogen peroxide in human trophoblast cells in vitro, by examining the changes in several markers implicated in oxidative stress interactions in the placenta. Trophoblast HTR-8/SVneo cells were preincubated with OLE at 10 and 100 µM and exposed to H2O2, as a model of oxidative stress. Protein and lipid peroxidation, as well as antioxidant enzymes’ activity, were determined spectrophotometrically, and DNA damage was evaluated by comet assay. iNOS protein expression was assessed by Western blot, while the mRNA expression of pro- and anti-apoptotic genes BAX and BCL2 and transcription factor NFE2L2, as well as cytokines IL-6 and TNF α were determined by qPCR. Oleuropein demonstrated cytoprotective effects against H2O2 in trophoblast cells by significantly improving the antioxidant status and preventing protein and lipid damage, as well as reducing the iNOS levels. OLE reduced the mRNA expression of IL-6 and TNF α, however, it did not influence the expression of NFE2L2 or the BAX/BCL2 ratio after H2O2 exposure. Oleuropein per se did not lead to any adverse effects in HTR-8/SVneo cells under the described conditions, confirming its safety in vitro. In conclusion, it significantly attenuated oxidative damage and restored antioxidant functioning, confirming its protective role in trophoblast. © 2023 by the authors

Oleuropein Attenuates Oxidative Stress in Human Trophoblast Cells

Olive-derived bioactive compound oleuropein was evaluated against damage induced by hydrogen peroxide in human trophoblast cells in vitro, by examining the changes in several markers implicated in oxidative stress interactions in the placenta. Trophoblast HTR-8/SVneo cells were preincubated with OLE at 10 and 100 µM and exposed to H2O2, as a model of oxidative stress. Protein and lipid peroxidation, as well as antioxidant enzymes’ activity, were determined spectrophotometrically, and DNA damage was evaluated by comet assay. iNOS protein expression was assessed by Western blot, while the mRNA expression of pro- and anti-apoptotic genes BAX and BCL2 and transcription factor NFE2L2, as well as cytokines IL-6 and TNF α were determined by qPCR. Oleuropein demonstrated cytoprotective effects against H2O2 in trophoblast cells by significantly improving the antioxidant status and preventing protein and lipid damage, as well as reducing the iNOS levels. OLE reduced the mRNA expression of IL-6 and TNF α, however, it did not influence the expression of NFE2L2 or the BAX/BCL2 ratio after H2O2 exposure. Oleuropein per se did not lead to any adverse effects in HTR-8/SVneo cells under the described conditions, confirming its safety in vitro. In conclusion, it significantly attenuated oxidative damage and restored antioxidant functioning, confirming its protective role in trophoblast. © 2023 by the authors

Galectin-1 Is Part of Human Trophoblast Invasion Machinery - A Functional Study In Vitro

Interactions of glycoconjugates with endogenous galectins, have been long proposed to participate in several reproductive processes including implantation. In human placenta gal-1, gal-3, gal-8, and gal-13 proteins are known to be present. Each of them has been proposed to play multiple functions, but so far no clear picture has emerged. We hypothesized that gal-1 participates in trophoblast invasion, and conducted Matrigel invasion assay using isolated cytotrophoblast from first trimester placenta and HTR-8/SVneo cell line to test it.<0.001) by Ox-gal-1 at 1 µg/ml. Both sets of results confirmed involvement of gal-1 in trophoblast invasion. Galectin profile of isolated cytotrophoblast and HTR-8/SVneo cells was established using RT-PCR and real-time PCR and found to consist of gal-1, gal-3 and gal-8 for both cell types. Only gal-1 was located at the trophoblast cell membrane, as determined by FACS analysis, which is consistent with the results of the functional tests.These findings qualify gal-1 as a member of human trophoblast cell invasion machinery

Steroid hormones modulate galectin-1 in the trophoblast HTR-8/SVneocell line

The effects of steroids on galectin-1 (gal-1) were studied in HTR-8/SVneo cells by immunocytochemistry, cell-based ELISA, the MTT proliferation test and the Matrigel TM invasion test. Dexamethasone (DEX), progesterone (PRG), and mifepristone (RU486) were used. Gal-1 was modulated in a steroid- and dose-dependent manner by DEX, which mildly but significantly stimulated production at low concentrations (0.1-10 nM), and inhibited it at 100 nM, while the effects of PRG and RU486 were opposite. HTR-8/SVneo cell invasion of Matrigel was significantly decreased in the presence of DEX and lactose. The obtained data support the proposed regulatory role of steroids in trophoblast gal-1 production