scholarly journals Characterization of the stress-inducing effects of homocysteine

1998 ◽  
Vol 332 (1) ◽  
pp. 213-221 ◽  
Author(s):  
P. Andrew OUTINEN ◽  
Sudesh K. SOOD ◽  
Patricia C. Y. LIAW ◽  
Kevin D. SARGE ◽  
Nobuyo MAEDA ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Stress Response ◽  
Heat Shock ◽  
Umbilical Vein ◽  
Mrna Levels ◽  
Stress Response Genes ◽  
Umbilical Vein Endothelial Cells ◽  
Shock Proteins

The mechanism by which homocysteine causes endothelial cell (EC) injury and/or dysfunction is not fully understood. To examine the stress-inducing effects of homocysteine on ECs, mRNA differential display and cDNA microarrays were used to evaluate changes in gene expression in cultured human umbilical-vein endothelial cells (HUVEC) exposed to homocysteine. Here we show that homocysteine increases the expression of GRP78 and GADD153, stress-response genes induced by agents or conditions that adversely affect the function of the endoplasmic reticulum (ER). Induction of GRP78 was specific for homocysteine because other thiol-containing amino acids, heat shock or H2O2 did not appreciably increase GRP78 mRNA levels. Homocysteine failed to elicit an oxidative stress response in HUVEC because it had no effect on the expression of heat shock proteins (HSPs) including HSP70, nor did it activate heat shock transcription factor 1. Furthermore homocysteine blocked the H2O2-induced expression of HSP70. In support of our findings in vitro, steady-state mRNA levels of GRP78, but not HSP70, were elevated in the livers of cystathionine β-synthase-deficient mice with hyperhomocysteinaemia. These studies indicate that the activation of stress response genes by homocysteine involves reductive stress leading to altered ER function and is in contrast with that of most other EC perturbants. The observation that homocysteine also decreases the expression of the antioxidant enzymes glutathione peroxidase and natural killer-enhancing factor B suggests that homocysteine could potentially enhance the cytotoxic effect of agents or conditions known to cause oxidative stress.

scholarly journals Identification and characterization of molecular interactions between mortalin/mtHsp70 and HSP60

2005 ◽  
Vol 391 (2) ◽  
pp. 185-190 ◽  
Author(s):  
Renu Wadhwa ◽  
Syuichi Takano ◽  
Kamaljit Kaur ◽  
Satoshi Aida ◽  
Tomoko Yaguchi ◽  
...  
Keyword(s):  
Heat Shock ◽  
Molecular Interactions ◽  
Heat Shock Protein 60 ◽  
Hsp60 Expression ◽  
Mitochondrial Hsp70 ◽  
Shock Proteins ◽  
First Time

Mortalin/mtHsp70 (mitochondrial Hsp70) and HSP60 (heat-shock protein 60) are heat-shock proteins that reside in multiple subcellular compartments, with mitochondria being the predominant one. In the present study, we demonstrate that the two proteins interact both in vivo and in vitro, and that the N-terminal region of mortalin is involved in these interactions. Suppression of HSP60 expression by shRNA (short hairpin RNA) plasmids caused the growth arrest of cancer cells similar to that obtained by suppression of mortalin expression by ribozymes. An overexpression of mortalin, but not of HSP60, extended the in vitro lifespan of normal fibroblasts (TIG-1). Taken together, this study for the first time delineates: (i) molecular interactions of HSP60 with mortalin; (ii) their co- and exclusive localizations in vivo; (iii) their involvement in tumorigenesis; and (iv) their functional distinction in pathways involved in senescence.

scholarly journals A Protective Role of Paeoniflorin in Fluctuant Hyperglycemia-Induced Vascular Endothelial Injuries through Antioxidative and Anti-Inflammatory Effects and Reduction of PKCβ1

2019 ◽  
Vol 2019 ◽  
pp. 1-11 ◽  
Author(s):  
Jing-Shang Wang ◽  
Ye Huang ◽  
Shuping Zhang ◽  
Hui-Jun Yin ◽  
Lei Zhang ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Protein Level ◽  
Umbilical Vein ◽  
Protective Role ◽  
Vascular Endothelial ◽  
Glucose Levels ◽  
Umbilical Vein Endothelial Cells

Hyperglycemia fluctuation is associated with diabetes mellitus (DM) complications when compared to persistent hyperglycemia. Previous studies have shown that paeoniflorin (PF), through its antiapoptosis, anti-inflammation, and antithrombotic properties, effectively protects against cardiovascular and cerebrovascular disease. However, the mechanism underlying the protection from PF against vascular injuries induced by hyperglycemia fluctuations remains poorly understood. Herein, we investigated the potential protective role of PF on human umbilical vein endothelial cells (HUVECs) subjected to intermittent glucose levels in vitro and in DM rats with fluctuating hyperglycemia in vivo. A remarkable increased apoptosis associated with elevated inflammation, increased oxidative stress, and high protein level of PKCβ1 was induced in HUVECs by intermittently changing glucose for 8 days, and PF recovered those detrimental changes. LY333531, a potent PKCβ1 inhibitor, and metformin manifested similar effects. Additionally, in DM rats with fluctuating hyperglycemia, PF protected against vascular damage as what has been observed in vitro. Taken together, PF attenuates the vascular injury induced by fluctuant hyperglycemia through oxidative stress inhibition, inflammatory reaction reduction, and PKCβ1 protein level repression, suggesting its perspective clinical usage.

Expression of heat shock genes of Neurospora crassa: effect of hyperthermia and other stresses on mRNA levels

1988 ◽  
Vol 66 (2) ◽  
pp. 81-92 ◽  
Author(s):  
Carol A. Curle ◽  
M. Kapoor
Keyword(s):  
Heat Shock ◽  
Neurospora Crassa ◽  
Sodium Arsenite ◽  
Gene Probe ◽  
Mrna Levels ◽  
Hsp 70 ◽  
Northern Blots ◽  
Stressed Cells ◽  
Shock Proteins

Neurospora crassa mycelium was heat shocked for intervals varying from 15–180 min. Heat shock mRNA was monitored by hybridization of Northern blots with the Drosophila hsp-70 gene probe and an inducible member of the yeast hsp-70 gene family, YG100. A 2.7 kilobase (kb) transcript, with homology to these two probes, was detected in cultures shocked for 15 min; its levels increased up to 60–90 min and declined thereafter. Sodium arsenite, too, induced the synthesis of this transcript. An additional, constitutively synthesized 2.4-kb transcript was revealed by hybridization with the yeast probe. The synthesis of this message was terminated during heat shock. Hybridization of Northern blots with the Drosophila actin gene probe demonstrated two size classes, 1.85 and 1.63 kb; the former decreased dramatically following heat shock. Recovery, as assessed by the disappearance of the 2.7-kb hsp-70-mRNA and restoration of the 1.85-kb actin message to the prestress levels, was essentially complete within 60 min of transfer to 28 °C. In vitro translations of RNA from stressed cells showed the heat shock messages to be stable and readily translatable. RNA of cells subjected to heat shock plus CdCl2 showed a higher content of messages for heat shock proteins of 70, 80, and 90 kilodaltons.

Impact of high CO2 levels on heat shock proteins during postharvest storage of table grapes at low temperature. Functional in vitro characterization of VVIHSP18.1

2018 ◽  
Vol 145 ◽  
pp. 108-116 ◽  
Author(s):  
Irene Romero ◽  
Ana C. Casillas-Gonzalez ◽  
Sergio J. Carrazana-Villalba ◽  
M. Isabel Escribano ◽  
Carmen Merodio ◽  
...  
Keyword(s):  
Heat Shock ◽  
Low Temperature ◽  
Heat Shock Proteins ◽  
High Co2 ◽  
Table Grapes ◽  
In Vitro Characterization ◽  
Co2 Levels ◽  
Shock Proteins

scholarly journals Up-regulation of thrombomodulin by activation of histamine H1-receptors in human umbilical-vein endothelial cells in vitro

1991 ◽  
Vol 276 (3) ◽  
pp. 739-743 ◽  
Author(s):  
K Hirokawa ◽  
N Aoki
Keyword(s):  
Endothelial Cells ◽  
Phorbol Esters ◽  
Umbilical Vein ◽  
Feedback Regulation ◽  
Mrna Levels ◽  
Human Umbilical Vein ◽  
H2 Antagonist ◽  
Histamine H1 Receptors ◽  
Umbilical Vein Endothelial Cells

Previous reports demonstrated that the expression of thrombomodulin (TM) in endothelial cells was modulated by various agents. Although TM was down-regulated by endotoxin or cytokines, up-regulation of TM was accomplished when endothelial cells were stimulated with unphysiologically high concentrations of cyclic AMP derivatives or tumour-promoting phorbol esters. We investigated the expression of TM in human umbilical-vein endothelial cells (HUVECs) by physiological substances that can be released into the bloodstream. Histamine (0.1-10 microM, 1-48 h) increased TM activity, TM antigen in cell lysates and TM mRNA levels, but 5-hydroxytryptamine and bradykinin had no effect. Enhancement of TM activity by histamine was completely blocked by the H1-selective antagonist pyrilamine, whereas the H2-antagonist cimetidine had no effect, showing that histamine up-regulates TM activity via H1-receptors on HUVECs. Enhanced TM activity by histamine and the resultant increase in protein C activation might play a role in a feedback regulation for prevention of vascular thrombosis.

scholarly journals Oxidative stress increases placental and endothelial cell activin A secretion

2007 ◽  
Vol 192 (3) ◽  
pp. 485-493 ◽  
Author(s):  
Stephen Mandang ◽  
Ursula Manuelpillai ◽  
Euan M Wallace
Keyword(s):  
Oxidative Stress ◽  
Umbilical Vein ◽  
Normal Pregnancy ◽  
Maternal Plasma ◽  
Activin A ◽  
Blood Monocytes ◽  
Wet Weight ◽  
Umbilical Vein Endothelial Cells ◽  
Circulating Levels

Circulating levels of activin A are significantly increased in women with preeclampsia when compared with those with a normal pregnancy. The mechanisms underlying these increased levels are unknown. We undertook these studies to explore whether oxidative stress might be the mechanism. We exposed trophoblast explants, human umbilical vein endothelial cells (HUVECs) and peripheral blood monocytes to oxidative stress in vitro using xanthine/xanthine oxidase (X/XO), measuring activin A and isoprostane in conditioned media and mRNA for activin βA in explants and HUVECs. We also measured isoprostane and activin A in serum from 21 women with preeclampsia and from 20 women with a normal pregnancy. Treatment with X/XO significantly increased 8-isoprostane production from placental explants, HUVECs and monocytes, indicative of oxidative stress, and significantly increased activin A output from placental explants (139.1 ± 27.4 per mg wet weight vs 322.9 ± 89.7 pg/ml per mg wet weight, P = 0.02) and from HUVECs (1.2 ± 0.2 vs 3.2 ± 1.8 ng/ml, P = 0.04). There was no effect on activin A output from monocytes. X/XO significantly increased βA mRNA in placental explants but not in HUVECs. Maternal plasma levels of 8-isoprostane and activin A were significantly higher in women with preeclampsia when compared with controls (333.8 ± 70 vs 176.3 ± 26.2 pg/ml, P = 0.04 and 49.5 ± 7 vs 13.1 ± 1.2 ng/ml, P < 0.001 respectively). In the women with preeclampsia, but not in those with a normal pregnancy, circulating levels of 8-isoprostane and activin A were significantly and positively correlated (r2 = 0.72; P < 0.001). These data suggest that oxidative stress may be one of the mechanisms underlying increased circulating activin A in preeclampsia.

Exertional heat injury and gene expression changes: a DNA microarray analysis study

2004 ◽  
Vol 96 (5) ◽  
pp. 1943-1953 ◽  
Author(s):  
Larry A. Sonna ◽  
C. Bruce Wenger ◽  
Scott Flinn ◽  
Holly K. Sheldon ◽  
Michael N. Sawka ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stress Response ◽  
Heat Shock ◽  
Heat Shock Proteins ◽  
Molecular Evidence ◽  
Peripheral Blood Mononuclear ◽  
Heat Injury ◽  
Shock Proteins

This study examined gene expression changes associated with exertional heat injury (EHI) in vivo and compared these changes to in vitro heat shock responses previously reported by our laboratory. Peripheral blood mononuclear cell (PBMC) RNA was obtained from four male Marine recruits (ages 17-19 yr) who presented with symptoms consistent with EHI, core temperatures ranging from 39.3 to 42.5°C, and elevations in serum enzymes such as creatine kinase. Controls were age- and gender-matched Marines from whom samples were obtained before and several days after an intense field-training exercise in the heat (“The Crucible”). Expression analysis was performed on Affymetrix arrays (containing ∼12,600 sequences) from pooled samples obtained at three times for EHI group (at presentation, 2-3 h after cooling, and 1-2 days later) and compared with control values (average signals from two chips representing pre- and post-Crucible samples). After post hoc filtering, the analysis identified 361 transcripts that had twofold or greater increases in expression at one or more of the time points assayed and 331 transcripts that had twofold or greater decreases in expression. The affected transcripts included sequences previously shown to be heat-shock responsive in PBMCs in vitro (including both heat shock proteins and non-heat shock proteins), a number of sequences whose changes in expression had not previously been noted as a result of in vitro heat shock in PBMCs (including several interferon-induced sequences), and several nonspecific stress response genes (including ubiquitin C and dual-specificity phosphatase-1). We conclude that EHI produces a broad stress response that is detectable in PBMCs and that heat stress per se can only account for some of the observed changes in transcript expression. The molecular evidence from these patients is thus consistent with the hypothesis that EHI can result from cumulative effects of multiple adverse interacting stimuli.

scholarly journals Parallel Isolation and Characterization of Porcine Smooth Muscle, Endothelial and Mesenchymal Stromal Cells for Bioengineering Applications

2021 ◽  
Author(s):  
Sara Morini ◽  
Iris Pla-Palacín ◽  
Pilar Sainz-Arnal ◽  
Natalia Sánchez-Romero ◽  
Maria Falceto ◽  
...  
Keyword(s):  
Smooth Muscle ◽  
Cultured Cells ◽  
Umbilical Vein ◽  
Building Blocks ◽  
Cell Types ◽  
Aortic Smooth Muscle ◽  
Isolation And Characterization ◽  
Umbilical Vein Endothelial Cells

Abstract There is significant interest in the pig as the animal model of choice for organ transplantation and the study of tissue engineering (TE) products and applications. Currently, efforts are being taken to bioengineer solid organs to reduce donor shortages for transplantation. For complex organs such as the lung, heart, and liver, the vasculature represents a fundamental feature. Thus, to generate organs with a functional vascular network, the different cells constituting the building blocks of the blood vessels should be procured. However, due to species' specificities, porcine cell isolation, expansion, and characterization are not entirely straightforward compared to human cell procurement. Here, we report the establishment of simple and suitable methods for the isolation and characterization of distinct porcine cells for bioengineering purposes.We successfully isolated, expanded and characterized porcine bone marrow-derived mesenchymal stromal (pBM-MSC), aortic smooth muscle (pASMC), and umbilical vein endothelial cells (pUVEC). We demonstrated that the three cell types showed specific immunophenotypical features. Moreover, we demonstrated that pBM-MSC could preserve their multipotency in vitro, and pUVEC were capable of maintaining their functionality in vitro.These cultured cells could be further expanded and represent a useful cellular tool for TE purposes (i.e., for recellularization approaches of vascularized organs or in vitro angiogenesis studies).

Characterization of Gastrin-Induced Proangiogenic Effects In vivo in Orthotopic U373 Experimental Human Glioblastomas and In vitro in Human Umbilical Vein Endothelial Cells

2004 ◽  
Vol 10 (24) ◽  
pp. 8250-8265 ◽  
Author(s):  
Florence Lefranc ◽  
Tatjana Mijatovic ◽  
Véronique Mathieu ◽  
Sandrine Rorive ◽  
Christine Decaestecker ◽  
...  
Keyword(s):  
Endothelial Cells ◽  
Umbilical Vein ◽  
Human Umbilical Vein ◽  
Umbilical Vein Endothelial Cells
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