Role of NO in cyclosporin nephrotoxicity: effects of chronic NO inhibition and NO synthases gene expression

1998 ◽  
Vol 274 (4) ◽  
pp. F791-F798 ◽  
Author(s):  
Norma A. Bobadilla ◽  
Gerardo Gamba ◽  
Edilia Tapia ◽  
Romeo García-Torres ◽  
Alexis Bolio ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Renal Cortex ◽  
Fold Increase ◽  
No Production ◽  
Renal Vasoconstriction ◽  
Cortical Level ◽  
Single Nephron ◽  
Enos Mrna ◽  
Proximal Tubular

The role of nitric oxide (NO) during cyclosporin renal vasoconstriction was evaluated by glomerular hemodynamic and histological changes produced by chronic NO synthesis inhibition and neuronal (nNOS), inducible (iNOS), and endothelial (eNOS) NO synthases mRNA expression in renal cortex and medulla. Uninephrectomized rats treated during 7 days with vehicle (Veh), cyclosporin A (CsA) 30 mg/kg, CsA + nitro-l-arginine methyl ester (l-NAME), and Veh +l-NAME (10 mg/dl) in the drinking water were studied. Increase in arterial pressure and afferent and efferent resistances, as well as decrease in glomerular plasma flow, ultrafiltration coefficient, and single-nephron glomerular filtration rate were significantly greater with CsA +l-NAME than with CsA alone. The increase in afferent resistance was higher with CsA +l-NAME than with Veh +l-NAME. In addition, glomerular thrombosis, proximal tubular vacuolization, and arteriolar thickening were more prominent. In renal cortex, eNOS mRNA expression exhibited a 2.7-fold increase in CsA, whereas, in medulla, nNOS and iNOS expression were lower in CsA than in Veh, while eNOS tended to increase. Our results support the hypothesis that NO synthesis is enhanced at cortical level during CsA nephrotoxicity, counterbalancing predominantly preglomerular vasoconstriction. Higher NO production could be the result of increased eNOS mRNA expression.

Abstract P196: Effect of Combined Oral Intake of Ginger Extract and Rice Vinegar on Blood Pressure with 2-kidney,1-clip Renovascular Hypertensive Rats

Hypertension ◽  
2017 ◽  
Vol 70 (suppl_1) ◽  
Author(s):  
Natsumi Saito ◽  
Yukiko Segawa ◽  
Saki Maruyama ◽  
Ayuna Yamaoka ◽  
Hiroko Hashimoto ◽  
...  
Keyword(s):  
Blood Pressure ◽  
Mrna Expression ◽  
Tap Water ◽  
Left Renal Artery ◽  
No Production ◽  
Sham Operation ◽  
Ginger Extract ◽  
Angiotensin Type 2 Receptor ◽  
Enos Mrna ◽  
Rice Vinegar

Objective: Ginger is widely used as traditional Asian herbal medicine. Ginger has the same pungent ingredient as chili and vanillyl. We showed that administration of capsaicin to renovascular hypertension (RH) model rats increased endothelial nitric oxide (NO) synthase (eNOS) mRNA expression and NO production, and suppressed blood pressure (BP). Traditionally in Japan, ginger is pickled and eaten. Ginger and vinegar each are supposed to have an effect of suppressing an increase in BP in RH rats. The aim of this study was to investigate the effect. Method: Male Sprague-Dawley rats (6wks) were treated with sham operation (SHAM) as controls or clipping the left renal artery (2K1C) as RH model. After surgery, the rats started receiving a control diet (C) or a diet with 0.08% (w/w) of Ginger Extract (GE) for 6 weeks, and a tap water (W) or a water with 4.5% (v/v) rice vinegar (V). The systolic BP (SBP) was measured by a tail-cuff method every week. At the end of the protocol, the mean arterial BP (MAP) was measured under anesthesia. Then, the aortas were removed for extracting mRNA. mRNA for angiotensin type 2 receptor (AT 2 ) and eNOS was evaluated by real-time RT-PCR. Results: Through the experiment period, SBP was significantly effects in time, model (SHAM vs 2K1C), diet (C vs GE) , timeхanimal ( P <0.001, each) and water (W vs V) ( P <0.05). At the end of the protocol, 2K1C-C+W was higher in SBP than SHAM-C+W (176 ± 6 vs 138 ± 1 mmHg, P <0.05). 2K1C-GE+W showed lower SBP (150 ± 2 mmHg) than -C+W ( P <0.05). SBP was not significantly different in 2K1C-GE+V (149 ± 4mmHg) from in -GE+W. The observations in MAP were similar to those in SBP. AT 2 R mRNA expression showed significant effects in model ( P <0.05) : the mRNA in 2K1C-C+W (0.9 ± 0.2) was significantly greater than in SHAM-C+W (0.4 ± 0.1) ( P <0.05). There were no significant differences among the 2K1Cs: -C+W, -C+V (0.9 ± 0.1), -GE+W (0.8 ± 0.1) and -GE+V (0.9 ± 0.2). eNOS mRNA expression showed significant effects only in diet (CTL vs GE, P <0.05), but not in water and any interactions. Conclusion: Continuous ingestion of GE and V may suppress BP increase in 2K1C, respectively. Simultaneous ingestion of GE and V showed no enhanced effects compared to GE or V solo ingestion in 2K1C. The roles of eNOS and AT 2 R in the mechanism did not become clear in this study.

Shear stress induces eNOS mRNA expression and improves endothelium-dependent dilation in senescent soleus muscle feed arteries

2005 ◽  
Vol 98 (3) ◽  
pp. 940-946 ◽  
Author(s):  
Christopher R. Woodman ◽  
Elmer M. Price ◽  
M. Harold Laughlin
Keyword(s):  
Shear Stress ◽  
Mrna Expression ◽  
Soleus Muscle ◽  
Low Flow ◽  
No Production ◽  
Fischer 344 ◽  
Enos Mrna ◽  
Series Of Experiments ◽  
Endothelial Nitric Oxide ◽  
Feed Arteries

We tested the hypothesis that increased intraluminal shear stress induces endothelial nitric oxide (NO) synthase (eNOS) mRNA expression and improves endothelium-dependent dilation in senescent soleus muscle feed arteries (SFA) by increasing NO production. SFA were isolated from young (4 mo) and old (24 mo) male Fischer 344 rats and cannulated with two resistance-matched glass micropipettes. SFA were exposed to no flow (NF), low flow (LF), intermediate flow (IF), or high flow (HF) for 4 h. Mean intraluminal shear stress ranged from 0 to 82 dyn/cm2. At the end of the 4-h treatment period, eNOS mRNA expression was assessed in each SFA. eNOS mRNA expression was significantly lower in old NF SFA than in young NF SFA. In old SFA, eNOS mRNA expression was induced by IF (+154%) and HF (+136%), resulting in a level of expression that was not different from that of young SFA. In a separate series of experiments, SFA were pretreated with NF or HF for 4 h, and endothelial function was assessed by examining vasodilator responses to ACh. ACh-induced dilation was less in old NF SFA than young NF SFA. Pretreatment with HF improved ACh-induced dilation in old SFA such that the response was similar to that of young SFA. In the presence of Nω-nitro-l-arginine to inhibit NOS, ACh-induced dilation was inhibited in old HF SFA such that the response was no longer greater than that of old NF SFA. These results indicate that increased intraluminal shear stress induces eNOS mRNA expression and improves endothelium-dependent dilation in senescent SFA by increasing NO production.

Measurement of nephron filtration in the dog: role of proximal intratubular pressure

1981 ◽  
Vol 241 (3) ◽  
pp. F238-F243
Author(s):  
D. A. Hartupee ◽  
A. H. Gillies ◽  
F. G. Knox
Keyword(s):  
Proximal Tubule ◽  
Filtration Rate ◽  
Free Flow ◽  
Nephron Filtration Rate ◽  
Intratubular Pressure ◽  
Single Nephron Filtration Rate ◽  
Single Nephron ◽  
Proximal Tubular ◽  
Pressure Controlled

Previous studies concerning the measurement of single nephron filtration rate have shown that collections of proximal tubular fluid, in which an oil drop is held in a constant position, do not affect intratubular pressure in the early proximal tubule in the hydropenic rat. Since intratubular pressures are higher in the dog than the rat, we investigated the effect of position-controlled collections on proximal pressure and single nephron filtration rate (SNGFR) in the dog. During position-controlled collections, early proximal pressure fell 5.8 +/- 0.9 mmHg and SNGFR was 76.3 +/- 5.3 nl/min. During proximal re-collections, in which proximal pressure was maintained near the free-flow value using a long immobile oil block, SNGFR was significantly less, 44.4 +/- 5.5 nl/min. For each micropunctured kidney, SNGFR was also estimated by dividing GFR by the number of glomeruli (mean, 5.4 +/- 0.5 X 10(5)). Estimated SNGFR (50.9 +/- 6.3 nl/min) was not significantly different from pressure-controlled SNGFR but was significantly less than position-controlled SNGFR. Accordingly, in the dog, early proximal pressure decreases during position-controlled collection of proximal tubular fluid, resulting in an overestimation of SNGFR. This artifact can be avoided by controlling the intratubular pressure during collection of tubular fluid.

Endothelial nitric oxide production during in vitro simulation of external limb compression

2002 ◽  
Vol 282 (6) ◽  
pp. H2066-H2075 ◽  
Author(s):  
Guohao Dai ◽  
Olga Tsukurov ◽  
Michael Chen ◽  
Jonathan P. Gertler ◽  
Roger D. Kamm
Keyword(s):  
Nitric Oxide ◽  
Mrna Expression ◽  
Umbilical Vein ◽  
Control Group ◽  
No Production ◽  
Vein Thrombosis ◽  
Enos Mrna ◽  
Umbilical Vein Endothelial Cells ◽  
Endothelial Nitric Oxide

External pneumatic compression (EPC) is effective in preventing deep vein thrombosis (DVT) and is thought to alter endothelial thromboresistant properties. We investigated the effect of EPC on changes in nitric oxide (NO), a critical mediator in the regulation of vasomotor and platelet function. An in vitro cell culture system was developed to simulate flow and vessel collapse conditions under EPC. Human umbilical vein endothelial cells were cultured and subjected to tube compression (C), pulsatile flow (F), or a combination of the two (FC). NO production and endothelial nitric oxide synthase (eNOS) mRNA expression were measured. The data demonstrate that in the F and FC groups, there is a rapid release of NO followed by a sustained increase. NO production levels in the F and FC groups were almost identical, whereas the C group produced the same low amount of NO as the control group. Conditions F and FC also upregulate eNOS mRNA expression by a factor of 2.08 ± 0.25 and 2.11 ± 0.21, respectively, at 6 h. Experiments with different modes of EPC show that NO production and eNOS mRNA expression respond to different time cycles of compression. These results implicate enhanced NO release as a potentially important factor in the prevention of DVT.

scholarly journals L-sepiapterin restores SLE serum-induced markers of endothelial function in endothelial cells

2019 ◽  
Vol 6 (1) ◽  
pp. e000294 ◽  
Author(s):  
Joy N Jones Buie ◽  
Dorea Pleasant Jenkins ◽  
Robin Muise-Helmericks ◽  
Jim C Oates
Keyword(s):  
Nitric Oxide ◽  
Endothelial Cells ◽  
Mrna Expression ◽  
Neutrophil Migration ◽  
Basal Medium ◽  
Neutrophil Adhesion ◽  
No Production ◽  
Functional Changes ◽  
Enos Mrna ◽  
The Impact

ObjectiveSLE serves as an independent risk factor` for endothelial dysfunction (ED) not explained by Framingham risk factors. We sought to understand the development of SLE-induced ED on a cellular level in order to develop strategies aimed at reversing cellular abnormalities. This study assessed the impact of SLE patient serum on endothelial nitric oxide synthase (eNOS), nitric oxide (NO) production and functional changes in the cell.MethodsHuman umbilical vein endothelial cells (HUVECs) cultured in serum of either SLE (n=25) or healthy patients (n=14) or endothelial basal medium 2 (EBM-2) culture media supplemented with fetal bovine serum with or without L-sepiapterin were used for our studies. We applied the fluorescent probe DAF-FM diacetate for intracellular NO detection using flow cytometry. Total RNA isolates were analysed using reverse transcription PCR for eNOS mRNA expression. Oxygen consumption rate was determined using seahorse analysis. Neutrophil adhesion and migration were determined using a calcein AM microscopy assay.ResultsThe mRNA expression of eNOS was increased in SLE cultured HUVECs compared with healthy control (p<0.05). The SLE eNOS mRNA level correlated with SLE patient age (p=0.008); however, this trend was not observed with healthy patients. SLE serum reduced NO production in HUVECs compared with EBM-2 cultured cells (p<0.05). Co-treatment of endothelial cells with L-sepiapterin preserved HUVEC capacity to produce NO in SLE conditions (p<0.01). SLE serum enhanced neutrophil migration (p<0.01) but not neutrophil adhesion compared with healthy controls. The bioenergetic health index was not different.ConclusionsSLE likely causes disruption of endothelial cell eNOS function and NO modulated pathways.

Modulation of factor VII levels by intron 7 polymorphisms: population and in vitro studies

Blood ◽  
2000 ◽  
Vol 95 (11) ◽  
pp. 3423-3428 ◽  
Author(s):  
Mirko Pinotti ◽  
Raffaella Toso ◽  
Domenico Girelli ◽  
Debora Bindini ◽  
Paolo Ferraresi ◽  
...  
Keyword(s):  
Mrna Expression ◽  
Fold Increase ◽  
Factor Vii ◽  
The Novel ◽  
Expression Studies ◽  
Sequence Variations ◽  
Fvii Deficiency ◽  
The Individual

Abstract Previous studies have established that factor VII gene (F7) polymorphisms (5′F7 and R353Q) contribute about one-third of factor VII (FVII) level variation in plasma. However, F7 genotyping in patients with cardiovascular disease has produced conflicting results. Population and expression studies were used to investigate the role of intron 7 (IVS7 ) polymorphisms, including repeat and sequence variations, in controlling activated FVII (FVIIa) and antigen (FVIIag) levels. Genotype–phenotype studies performed in 438 Italian subjects suggested a positive relation between the IVS7 repeat number and FVII levels. The lowest values were associated with theIVS7 + 7G allele. The screening of 52 patients with mild FVII deficiency showed an 8-fold increase in frequency (8%) of this allele, and among heterozygotes for identical mutations, lower FVII levels were observed in the IVS7 + 7G carriers. This frequent genetic component participates in the phenotypic heterogeneity of FVII deficiency. The evaluation of the individual contribution of polymorphisms was assisted by the expression of each IVS7variant, as a minigene, in eukaryotic cells. The novel quantitative analysis revealed that higher numbers of repeats were associated with higher mRNA expression levels and that the IVS7 + 7Gallele, previously defined as a functionally silent polymorphism, was responsible for the lowest relative mRNA expression. Taken together, these findings indicate that the IVS7 polymorphisms contribute to the plasmatic variance of FVII levels via differential efficiency of mRNA splicing. These studies provide further elements to understand the control of FVII levels, which could be of importance to ensure the hemostatic balance under pathologic conditions.

Enhanced Adhesion, Migration and Pyk2 Expression in K562 Cells Following Imatinib Treatment

Blood ◽  
2012 ◽  
Vol 120 (21) ◽  
pp. 4426-4426
Author(s):  
Adelina Ovcharenko ◽  
Galit Granot ◽  
Jeniffer Park ◽  
Ofer Shpilberg ◽  
Pia Raanani
Keyword(s):  
Cell Adhesion ◽  
Targeted Therapy ◽  
Mrna Expression ◽  
Cell Line ◽  
Focal Adhesion ◽  
K562 Cell ◽  
K562 Cells ◽  
Fold Increase ◽  
And Migration

Abstract Abstract 4426 Background/Aims: Despite improved prognosis of CML patients with the use of imatinib (IM), its administration is associated with extramedullary disease (EMD) occurrence. We postulate that, like in the metastatic processes, changes in migration and adherence potential may enable leukemic cells to inhabit extramedullary sites. Focal adhesion complexes linking between extracellular matrix and the cell cytoskeleton are likely to play an important role in these processes. Pyk2 is a tyrosine kinase highly expressed in hematopoietic cells, localized to focal adhesion complexes, and known to participate in adhesion and migration processes. We have previously shown that Pyk2 participates in NB4 (an acute promyelocytic leukemia [APL] cell line) cells' adhesion and migration following exposure to the APL targeted therapy ATRA. We postulate that similar to the effect of ATRA on NB4 cells, IM being also a targeted therapy, might also be associated with enhanced adhesion and migration abilities of the CML cell line K562. Our objectives were to identify the effect of IM administration on pyk2 expression and on K562 cell adhesion and migration ability and to establish the role of these changes in treatment-associated EMD. Results: We found a 2.6-fold increase in pyk2 mRNA expression in K562 cells following exposure to IM. We also found that 30% of IM-treated K562 cells adhered to fibronectin (FN) compared to untreated cells having no adhesion ability. In addition, a 3-fold induction in migration was seen in K562 cells following treatment. Furthermore, K562 cells treated with IM demonstrated a 2-fold increase in invasion potential as compared to untreated cells. In order to assess whether Pyk2 is essential for IM-dependent adhesion and migration of K562 cells, these cells were infected with pyk2 specific shRNAs. While 30% of the non-infected NB4 cells adhered to FN following IM treatment, only 12% of the pyk2-shRNA–infected K562 cells exhibited adhesion potential (Pvalue<0.002). In addition, we witnessed over a 3-fold reduction in the ability of pyk2-shRNA–infected K562 cells to migrate following exposure to IM when compared to parental K562 cells. These data support the role of Pyk2 in IM-mediated adhesion and migration. Finally, we found that IM treatment induced an in-vivo increase in pyk2 mRNA expression level in leukocytes derived from 3 out of 5 CML patients studied. Conclusions: IM induces K562 cell adhesion, migration and invasion accompanied by increased pyk2 expression. Pyk2 is one of the key proteins regulating IM-induced cell migration and adhesion. Collectively our data suggest a critical role of Pyk2 in adhesion and migration initiated by the targeted therapy IM and a possible role in EMD development. These data support a common mechanism for the development of EMD in hematological malignancies treated by targeted therapies via pyk2 expression. Disclosures: No relevant conflicts of interest to declare.

scholarly journals Involvement of Nitric Oxide in Iodine Deficiency-Induced Microvascular Remodeling in the Thyroid Gland: Role of Nitric Oxide Synthase 3 and Ryanodine Receptors

Endocrinology ◽  
2014 ◽  
Vol 156 (2) ◽  
pp. 707-720 ◽  
Author(s):  
J. Craps ◽  
C. Wilvers ◽  
V. Joris ◽  
B. De Jongh ◽  
J. Vanderstraeten ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Blood Flow ◽  
Thyroid Gland ◽  
Mrna Expression ◽  
Iodine Deficiency ◽  
Ryanodine Receptors ◽  
No Production

Iodine deficiency (ID) induces microvascular changes in the thyroid gland via a TSH-independent reactive oxygen species-hypoxia inducible factor (HIF)-1α-vascular endothelial growth factor (VEGF) pathway. The involvement of nitric oxide (NO) in this pathway and the role of calcium (Ca2+) and of ryanodine receptors (RYRs) in NO synthase 3 (NOS3) activation were investigated in a murine model of goitrogenesis and in 3 in vitro models of ID, including primary cultures of human thyrocytes. ID activated NOS3 and the production of NO in thyrocytes in vitro and increased the thyroid blood flow in vivo. Using bevacizumab (a blocking antibody against VEGF-A) in mice, it appeared that NOS3 is activated upstream of VEGF-A. L-nitroarginine methyl ester (a NOS inhibitor) blocked the ID-induced increase in thyroid blood flow in vivo and NO production in vitro, as well as ID-induced VEGF-A mRNA and HIF-1α expression in vitro, whereas S-nitroso-acetyl-penicillamine (a NO donor) did the opposite. Ca2+ is involved in this pathway as intracellular Ca2+ flux increased after ID, and thapsigargin activated NOS3 and increased VEGF-A mRNA expression. Two of the 3 known mammalian RYR isoforms (RYR1 and RYR2) were shown to be expressed in thyrocytes. RYR inhibition using ryanodine at 10μM decreased ID-induced NOS3 activation, HIF-1α, and VEGF-A expression, whereas RYR activation with ryanodine at 1nM increased NOS3 activation and VEGF-A mRNA expression. In conclusion, during the early phase of TSH-independent ID-induced microvascular activation, ID sequentially activates RYRs and NOS3, thereby supporting ID-induced activation of the NO/HIF-1α/VEGF-A pathway in thyrocytes.

scholarly journals Renal nitric oxide production in rat pregnancy: role of constitutive nitric oxide synthases

2010 ◽  
Vol 299 (4) ◽  
pp. F830-F836 ◽  
Author(s):  
Cheryl A. Smith ◽  
Beth Santymire ◽  
Aaron Erdely ◽  
Vasuki Venkat ◽  
György Losonczy ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Renal Cortex ◽  
Soluble Fraction ◽  
Splice Variants ◽  
Normal Pregnancy ◽  
No Production ◽  
Protein Abundance ◽  
Functional Studies ◽  
Enos Protein

Functional studies show that increased renal nitric oxide (NO) mediates the renal vasodilation and increased glomerular filtration rate that occur during normal pregnancy. We investigated whether changes in the constitutive NO synthases (NOS), endothelial (eNOS) and neuronal (nNOS), were associated with the increased renal NO production in normal midterm pregnancy in the rat. In kidneys from midterm pregnant (MP: 11–13 days gestation), late-term pregnant (LP: 18–20 days gestation), and similarly aged virgin (V) rats, transcript and protein abundance for eNOS and the nNOSα and nNOSβ splice variants, as well as the rate of l-arginine-to-l-citrulline conversion, were determined as a measure of NOS activity. At MP, renal cortical abundance of the total eNOS protein and phosphorylated (Ser1177) eNOS was reduced, and l-arginine-to-l-citrulline conversion in the cortical membrane fraction was decreased; these declines were also seen in LP. There were no changes in the eNOS transcript. In contrast, l-arginine-to-l-citrulline conversion in the soluble fraction of renal cortex increased at MP and then declined at LP. This MP increase was ablated by S-methylthiocitrulline, a nNOS inhibitor. Using Western blotting, we did not detect a change in the protein abundance or transcript of the 160-kDa nNOSα, but protein abundance and transcript of the nNOSβ were increased at MP in cortex. Collectively, these studies suggest that the soluble nNOSβ is responsible for the increased renal cortical NO production during pregnancy.

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