scholarly journals Suppression of Nitric Oxide Implicated in the Protective Effect of Echinacoside on H2O2-Induced PC12 Cell Injury

2010 ◽  
Vol 5 (4) ◽  
pp. 1934578X1000500 ◽  
Author(s):  
Rong Kuang ◽  
Yiguo Sun ◽  
Xiaoxiang Zheng
Keyword(s):  
Nitric Oxide ◽  
Protective Effect ◽  
Pc12 Cells ◽  
Laser Scanning ◽  
Cell Injury ◽  
Inhibitory Effect ◽  
Pcr Analysis ◽  
Inos Mrna ◽  
Rt Pcr ◽  
Pheochromocytoma Cell

We investigated whether suppression of nitric oxide (NO) implicated in the protective effect of echinacoside (ECH), a phenylethanoid glycoside, on H2O2-induced injury to the rat pheochromocytoma cell line (PC12 cells). Data show that application of ECH to H2O2-injured PC12 cells (HIPCs) increased cell viability and decreased the necrotic ratio. Laser scanning confocal microscopic (LSCM) analysis suggested that ECH exerted an inhibitory effect on the formation of NO. In addition, RT-PCR analysis revealed that ECH down-regulated p65 and iNOS mRNA expressions in HIPCs. In summary, suppression of NO is related to the protective effect of ECH on HIPCs.

scholarly journals Expression patterns of endothelial and inducible nitric oxide synthase isoforms in corpora lutea of pseudopregnant rabbits at different luteal stages

2002 ◽  
Vol 173 (2) ◽  
pp. 285-296 ◽  
Author(s):  
C Boiti ◽  
D Zampini ◽  
G Guelfi ◽  
F Paolocci ◽  
M Zerani ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Expression Patterns ◽  
Physiological Role ◽  
Total Activity ◽  
Pcr Analysis ◽  
Inos Mrna ◽  
Corpora Lutea ◽  
Mrna Levels ◽  
Isolated Cells ◽  
Protein Levels

Total activity of nitric oxide (NO) synthase (NOS) and expression of both endothelial (eNOS) and inducible (iNOS) isoforms were examined in corpora lutea (CL) of rabbits across pseudopregnancy by quantitative RT-PCR analysis, Western blot and immunohistochemistry. CL were collected at early- (day 4), mid- (day 9) and late- (day 13) luteal phases of pseudopregnancy. The PCR product of rabbit luteal eNOS was cloned and its direct sequence exhibited 90% homology with those of other species. The steady-state mRNA levels encoding eNOS remained fairly constant throughout both early- and mid-luteal stages of pseudopregnancy but dropped almost to half (P</=0.05) by day 13. By contrast, luteal eNOS proteins increased 2-fold (P</=0.05) from the early- to late-luteal phase. Independently of CL age, iNOS mRNA was very poorly expressed while protein levels gradually declined from the early- to late-luteal stage. Intense eNOS-like immunoreactivity was detected in large luteal cells, while iNOS staining was targeted to a few, isolated cells, probably macrophages. Basal NOS activity was greater in day 4 CL than in both day 9 and day 13 CL. These data are the first to characterize in rabbit CL the temporal expression patterns of NOS isoforms across different luteal stages of pseudopregnancy and, collectively, suggest the existence of an expressional control for this constitutive isoform, which might have a physiological role in regulating CL function during development.

scholarly journals Inhibitory Effect of Essential Oil from Fructus Alpiniae zerumbet on Endothelial-to-Mesenchymal Transformation Induced by TGF-β1 and down-regulating KLF4

2020 ◽  
Author(s):  
shuang zhao ◽  
Li He ◽  
Mei Huang ◽  
Meng xin Tu ◽  
Xiang chun Shen ◽  
...  
Keyword(s):  
Protective Effect ◽  
Protein Interactions ◽  
Umbilical Vein ◽  
Histone H3 ◽  
Inhibitory Effect ◽  
Pcr Analysis ◽  
Protective Effects ◽  
Migration Ability ◽  
Umbilical Vein Endothelial Cells ◽  
Mesenchymal Transformation

Abstract Background: Endothelial Mesenchymal Transformation (EndMT) contributes to the development of cardiovascular disease. Krüpple factor 4 (KLF4) is a zinc finger transcription factor whose N-terminus can recruit acetyltransferase to promote histone acetylation, thereby affecting the transcription activation of downstream genes. Our previous studies have shown that EOFAZ has protective effects on HUVECs oxidative stress induced by TGF-β1. However, whether EOFAZ has a protective effect on EndMT induced by TGF-β1 and whether it is related to the regulation of downstream signals by KLF4 has not yet been elucidated.Methods: The protective effects of EOFAZ were evaluated in TGF-β1-treated EndMT in Human umbilical vein endothelial cells (HUVECs). Cell mobility was evaluated by wound-healing, transwell assays and angiogenesis experiment. Western blot analysis, Quantitative real-time PCR analysis (qRT-PCR) and immunofluorescence staining were utilized to determine the expression of endothelial and mesenchymal markers , KLF4, Histone 3 acetylation and Notch/Snail signaling axis. Small interfering RNA (siRNA) and adenovirus infection were used to determine the effciency of KLF4 inhibition and overexpression. Immunoprecipitation experiments were performed to analyze protein interactions.Results: We reported that EOFAZ has a protective effect on EndMT induced by TGF-β1. Deletion of KLF4 inhibited EndMT induced by TGF-β1 in HUVECs. EOFAZ pretreatment and KLF4 knockout reduced the migration ability of HUVECs , and increased endothelial markers accompanied by decreased mesenchymal markers, meanwhile caused the change of Notch/Snail signal axis. In addition, TGF-β1 upregulated the expression of KLF4, while the high expression of KLF4 promoted the acetylation of histone H3, and there was a protein interaction between the acetylated histone H3 and KLF4. Conclusions: These results suggest that TGF-β1 may promote the acetylation of histone H3 and activate the transduction of Notch/Snail signal axis by up-regulating the expression of KLF4, which may induce EndMT and this effect may be reversed by EOFAZ. Therefore, EOFAZ may inhibit EndMT induced by TGF-β1 by down-regulating KLF4 expression.

scholarly journals RNA-Seq Based Transcriptome Analysis of Ethanol Extract of Saffron Protective Effect Against Corticosterone-Induced PC12 Cell Injury

2021 ◽  
Author(s):  
Xi Chen ◽  
Ting Yang ◽  
Cong' en Zhang ◽  
Zhijie Ma
Keyword(s):  
Protective Effect ◽  
Pc12 Cells ◽  
Pc12 Cell ◽  
Cell Injury ◽  
Ethanol Extract ◽  
Mapk Signaling ◽  
Cell Counting ◽  
Rna Seq ◽  
Traditional Chinese Herbal Medicine ◽  
Water Test

Abstract Background Saffron is a traditional Chinese herbal medicine, which is typically used in clinical to regulate anxiety, tension, and other depression-related conditions. The study aimed to explore the neuroprotective effect of ethanol extract of saffron (EES) on corticosterone (CORT)-induced injury in PC12 cells and further explored its potential mechanism. Methods The authenticity of saffron and the active components of EES were identified by a water test and ultra-performance liquid chromatography-time of flight mass spectrometry system. The screening of cytotoxicity for PC12 cells was incubated with EES in different concentrations for 24 h, and the protective efficacy of EES on CORT (500 µM) induced PC12 cell injury, cell viability was assessed by Cell Counting Kit-8 (CCK-8) assay. The DEGs of EES-protected PC12 cells were analyzed using the RNA-seq method, and the results were analyzed for GO and KEGG enrichment. The results of RNA-seq were verified by qPCR analysis. Results The saffron was initially identified as authentic in the water test and 10 compounds were identified by UPLC-MS. The results of CCK-8 demonstrated that EES at concentrations above 640 µg/mL exerted a certain cytotoxic effect, and PC12 cells pretreated with EES (20, 40, and 80 µg/mL) significantly reversed the 500 µM CORT-induced cell death. RNA-seq analysis showed that EES regulated 246 differential genes, which were mainly enriched in the MAPK signaling pathway. Dusp5, Dusp6, Gadd45b, Gadd45G, and Pdgfc were further validated by qPCR. Experimental data showed that the results of qPCR were consistent with RNA-seq. Conclusions These findings provide an innovative understanding of the molecular mechanism of the protective effect of EES on PC12 cells at the molecular transcription level, and the above molecules may be potential novel targets for antidepressant treatment.

Hydrogen peroxide downregulates IL-1-driven mesangial iNOS activity: implications for glomerulonephritis

1997 ◽  
Vol 272 (6) ◽  
pp. F721-F728 ◽  
Author(s):  
E. A. Jaimes ◽  
K. A. Nath ◽  
L. Raij
Keyword(s):  
Nitric Oxide ◽  
Hydrogen Peroxide ◽  
Mesangial Cells ◽  
Interleukin 1 ◽  
Inhibitory Effect ◽  
Inos Mrna ◽  
No Production ◽  
Glomerular Injury ◽  
Inos Activity ◽  
Oxide Synthase

In glomerulonephritides, autacoids such as nitric oxide (NO), reactive oxygen species, and prostanoids are produced in increased amounts in response to cytokines such as interleukin-1 (IL-1). These autacoids influence the expression of glomerular injury by their direct as well as interactive actions. We studied the effect of hydrogen peroxide (H2O2) on NO production in rat mesangial cells. We demonstrate that transient exposure of mesangial cells to H2O2 prior to sustained exposure to IL-1 decreased extracellular accumulation of NO2/NO3 and cellular guanosine 3,'5'-cyclic monophosphate (cGMP) content. H2O2 markedly impaired inducible nitric oxide synthase (iNOS) activity induced by IL-1 directly measured by the conversion of L-[14C]arginine to L-[14C]citrulline. Such impairment in iNOS activity was accompanied by a parallel reduction in iNOS protein abundance but not by a reduced expression of iNOS mRNA. The inhibitory effect of H2O2 on NOS activity was further supported by peroxide-induced impairment in IL-1-driven, NO-dependent synthesis of prostaglandin E2. Our studies thus provide the first direct evidence of a posttranscriptional inhibitory effect of H2O2 on iNOS activity. Additionally, our studies uncover the existence of a previously unrecognized effect of H2O2 on the production of NO that may exert influence on the severity of glomerular injury during glomerular inflammation.

Hesperidin protects against cyclophosphamide-induced hepatotoxicity by upregulation of PPARγ and abrogation of oxidative stress and inflammation

2014 ◽  
Vol 92 (9) ◽  
pp. 717-724 ◽  
Author(s):  
Ayman M. Mahmoud
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Protective Effect ◽  
Liver Lipid ◽  
Inos Mrna ◽  
Peroxisome Proliferator ◽  
Dependent Manner ◽  
Protective Effects ◽  
Peroxisome Proliferator Activated Receptor ◽  
Intraperitoneal Dose

The most important reason for the non-approval and withdrawal of drugs by the Food and Drug Administration is hepatotoxicity. Therefore, this study was undertaken to evaluate the protective effects of hesperidin against cyclophosphamide (CYP)-induced hepatotoxicity in Wistar rats. The rats received a single intraperitoneal dose of CYP of 200 mg/kg body mass, followed by treatment with hesperidin, orally, at doses of 25 and 50 mg/kg for 11 consecutive days. CYP induced hepatic damage, as evidenced by the significantly elevated levels of serum pro-inflammatory cytokines, serum transaminases, liver lipid peroxidation, and nitric oxide. As a consequence, there was reduced glutathione content, and the activities of the antioxidant enzymes superoxide dismutase, catalase, and glutathione peroxidase, were markedly reduced. In addition, CYP administration induced a considerable downregulation of peroxisome proliferator activated receptor gamma (PPARγ) and upregulation of nuclear factor-kappa B (NF-κB) and inducible nitric oxide synthase (iNOS) mRNA expression. Hesperidin, in a dose-dependent manner, rejuvenated the altered markers to an almost normal state. In conclusion, hesperidin showed a potent protective effect against CYP-induced oxidative stress and inflammation leading to hepatotoxicity. The study suggests that hesperidin exerts its protective effect against CYP-induced hepatotoxicity through upregulation of hepatic PPARγ expression and abrogation of inflammation and oxidative stress.

scholarly journals Interleukin-13 inhibits inducible nitric oxide synthase expression in human mesangial cells

1996 ◽  
Vol 313 (2) ◽  
pp. 641-646 ◽  
Author(s):  
Marta SAURA ◽  
Rosa MARTÍNEZ-DALMAU ◽  
Adrian MINTY ◽  
Dolores PÉREZ-SALA ◽  
Santiago LAMAS
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Mesangial Cells ◽  
Inhibitory Effect ◽  
Northern Analysis ◽  
Inos Mrna ◽  
Dependent Manner ◽  
Human Mesangial Cells ◽  
Oxide Synthase ◽  
Stimulation Of

The synthesis of nitric oxide in inflammatory situations requires the expression of an inducible isoform of nitric oxide synthase (iNOS). Human mesangial cells (HMC) express an iNOS enzyme after exposure to multiple co-stimuli. In this study we have observed that while tumour necrosis factor-α, interleukin (IL)-1β, interferon-γ and bacterial lipopolysaccharide (LPS) were unable to significantly induce NO synthesis when used alone, they induced an evident stimulation of NO synthesis when used in various combinations. A mixture of the three cytokines (CM) and LPS resulted in a 10-15-fold stimulation of NO synthesis over control values which started to be significant after 16 h. The addition of IL-13, a cytokine with anti-inflammatory properties, inhibited CM/LPS-induced NO synthesis in a concentration-dependent manner. A marked inhibitory effect (60-65%) could be observed when HMC were treated with IL-13 (10 ng/ml) 24 h before, at the same time as, or even 4 h after the addition of CM/LPS. This inhibitory effect was still significant (25%) when IL-13 was added 16 h after CM/LPS. Northern analysis showed that IL-13-mediated iNOS inhibition was closely correlated with the suppression of iNOS mRNA expression. These results identify IL-13 as a powerful regulatory tool for the inhibition of NO synthesis in human cells, a property which may be pathophysiologically relevant in NO-related inflammatory processes.

scholarly journals Chorein Sensitive Arrangement of Cytoskeletal Architecture

2015 ◽  
Vol 37 (1) ◽  
pp. 399-408 ◽  
Author(s):  
Sabina Honisch ◽  
Shuchen Gu ◽  
Jennifer Müller vom Hagen ◽  
Saad Alkahtani ◽  
Abdullah A. Al Kahtane ◽  
...  
Keyword(s):  
Laser Scanning ◽  
Actin Polymerization ◽  
Human Fibroblasts ◽  
Cell Types ◽  
Cytoskeletal Proteins ◽  
Laser Scanning Microscopy ◽  
Pcr Analysis ◽  
Loss Of Function ◽  
Rt Pcr ◽  
Behavioral Abnormalities

Background/Aims: Chorein is a protein expressed in various cell types. Loss of function mutations of the chorein encoding gene VPS13A lead to chorea-acanthocytosis, an autosomal recessive genetic disease characterized by movement disorder and behavioral abnormalities. Recent observations revealed that chorein is a powerful regulator of actin cytoskeleton in erythrocytes, platelets, K562 and endothelial HUVEC cells. Methods: In the present study we have used Western blotting to study actin polymerization dynamics, laser scanning microscopy to evaluate in detail the role of chorein in microfilaments, microtubules and intermediate filaments cytoskeleton architecture and RT-PCR to assess gene transcription of the cytoskeletal proteins. Results: We report here powerful depolymerization of actin microfilaments both, in erythrocytes and fibroblasts isolated from chorea-acanthocytosis patients. Along those lines, morphological analysis of fibroblasts from chorea-acanthocytosis patients showed disarranged microtubular network, when compared to fibroblasts from healthy donors. Similarly, the intermediate filament networks of desmin and cytokeratins showed significantly disordered organization with clearly diminished staining in patient's fibroblasts. In line with this, RT-PCR analysis revealed significant downregulation of desmin and cytokeratin gene transcripts. Conclusion: Our results provide for the first time evidence that defective chorein is accompanied by significant structural disorganization of all cytoskeletal structures in human fibroblasts from chorea-acanthocytosis patients.

Modulation of Nitric Oxide Production by Tetracyclines and Chemically Modified Tetracyclines

1998 ◽  
Vol 12 (1) ◽  
pp. 126-130 ◽  
Author(s):  
E. Cillari ◽  
S. Milano ◽  
P. D'Agostino ◽  
G. Di Bella ◽  
M. La Rosa ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Inhibitory Effect ◽  
Inos Mrna ◽  
Mrna Accumulation ◽  
Chemically Modified ◽  
No Formation ◽  
Oxide Synthase ◽  
Chemically Modified Tetracyclines ◽  
Dose Dependent Increase ◽  
Dose Dependent

Chemically modified tetracyclines (CMTs) dose-dependently decreased inducible nitric oxide synthase (iNOS) and, consequently, nitric oxide (NO) formation by the lipopolysaccharide (LPS)-stimulated J774 line. The inhibitory effect was due to a specific reduction in the iNOS protein content in the cells, as attested by Western blot analysis and by the inhibition of iNOS mRNA accumulation. Furthermore, CMTs cause a dose-dependent increase in cell death in the J774 line mediated by the NO-independent apoptotic mechanism.

scholarly journals Inhibition of CDKS by roscovitine suppressed LPS-induced ·NO production through inhibiting NFκB activation and BH4 biosynthesis in macrophages

2009 ◽  
Vol 297 (3) ◽  
pp. C742-C749 ◽  
Author(s):  
Jianhai Du ◽  
Na Wei ◽  
Tongju Guan ◽  
Hao Xu ◽  
Jianzhong An ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Inflammatory Diseases ◽  
Nuclear Factor Κb ◽  
Inhibitory Effect ◽  
Peritoneal Macrophages ◽  
Inos Mrna ◽  
No Production ◽  
Iκb Kinase ◽  
Cox 2 ◽  
Oxide Synthase

In inflammatory diseases, tissue damage is critically associated with nitric oxide (·NO) and cytokines, which are overproduced in response to cellular release of endotoxins. Here we investigated the inhibitory effect of roscovitine, a selective inhibitor of cyclin-dependent kinases (CDKs) on ·NO production in mouse macrophages. In RAW264.7 cells, we found that roscovitine abolished the production of ·NO induced by lipopolysaccharide (LPS). Moreover, roscovitine significantly inhibited LPS-induced inducible nitric oxide synthase (iNOS) mRNA and protein expression. Our data also showed that roscovitine attenuated LPS-induced phosphorylation of IκB kinase β (IKKβ), IκB, and p65 but enhanced the phosphorylation of ERK, p38, and c-Jun NH2-terminal kinase (JNK). In addition, roscovitine dose dependently inhibited LPS-induced expression of cyclooxygenase-2 (COX)-2, IL-1β, and IL-6 but not tumor necrosis factor (TNF)-α. Tetrahydrobiopterin (BH4), an essential cofactor for iNOS, is easily oxidized to 7,8-dihydrobiopterin (BH2). Roscovitine significantly inhibited LPS-induced BH4 biosynthesis and decreased BH4-to-BH2 ratio. Furthermore, roscovitine greatly reduced the upregulation of GTP cyclohydrolase-1 (GCH-1), the rate-limiting enzyme for BH4 biosynthesis. Using other CDK inhibitors, we found that CDK1, CDK5, and CDK7, but not CDK2, significantly inhibited LPS-induced ·NO production in macrophages. Similarly, in isolated peritoneal macrophages, roscovitine strongly inhibited ·NO production, iNOS, and COX-2 upregulation, activation of NFκB, and induction of GCH-1 by LPS. Together, our data indicate that roscovitine abolishes LPS-induced ·NO production in macrophages by suppressing nuclear factor-κB activation and BH4 biosynthesis, which might be mediated by CDK1, CDK5, and CDK7. Our results also suggest that roscovitine may inhibit inflammation and that CDKs may play important roles in the mechanisms by which roscovitine attenuates inflammation.

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