scholarly journals L-Arginine Enhances Protein Synthesis by Phosphorylating mTOR (Thr 2446) in a Nitric Oxide-Dependent Manner in C2C12 Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-13 ◽  
Author(s):  
Ruxia Wang ◽  
Hongchao Jiao ◽  
Jingpeng Zhao ◽  
Xiaojuan Wang ◽  
Hai Lin
Keyword(s):  
Nitric Oxide ◽  
Protein Synthesis ◽  
Culture Media ◽  
Muscle Protein ◽  
Muscle Protein Synthesis ◽  
C2c12 Cells ◽  
Dependent Manner ◽  
No Donor ◽  
Nos Inhibitor ◽  
Oxide Synthase

Muscle atrophy may arise from many factors such as inactivity, malnutrition, and inflammation. In the present study, we investigated the stimulatory effect of nitric oxide (NO) on muscle protein synthesis. Primarily, C2C12 cells were supplied with extra L-arginine (L-Arg) in the culture media. L-Arg supplementation increased the activity of inducible nitric oxide synthase (iNOS), the rate of protein synthesis, and the phosphorylation of mTOR (Thr 2446) and p70S6K (Thr 389). L-NAME, an NOS inhibitor, decreased NO concentrations within cells and abolished the stimulatory effect of L-Arg on protein synthesis and the phosphorylation of mTOR and p70S6K. In contrast, SNP (sodium nitroprusside), an NO donor, increased NO concentrations, enhanced protein synthesis, and upregulated mTOR and p70S6K phosphorylation, regardless of L-NAME treatment. Blocking mTOR with rapamycin abolished the stimulatory effect of both L-Arg and SNP on protein synthesis and p70S6K phosphorylation. These results indicate that L-Arg stimulates protein synthesis via the activation of the mTOR (Thr 2446)/p70S6K signaling pathway in an NO-dependent manner.

Inducible nitric oxide synthase augments injury elicited by oxidative stress in rat cardiac myocytes

1998 ◽  
Vol 274 (1) ◽  
pp. C245-C252 ◽  
Author(s):  
Junsuke Igarashi ◽  
Masashi Nishida ◽  
Shiro Hoshida ◽  
Nobushige Yamashita ◽  
Hiroaki Kosaka ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Nitric Oxide ◽  
Cardiac Myocytes ◽  
Myocardial Injury ◽  
No Production ◽  
Dependent Manner ◽  
Concentration Dependent Manner ◽  
No Donor ◽  
Oxide Synthase ◽  
Gpx Activity

The effects of nitric oxide (NO) produced by cardiac inducible NO synthase (iNOS) on myocardial injury after oxidative stress were examined. Interleukin-1β induced cultured rat neonatal cardiac myocytes to express iNOS. After induction of iNOS,l-arginine enhanced NO production in a concentration-dependent manner. Glutathione peroxidase (GPX) activity in myocytes was attenuated by elevated iNOS activity and by an NO donor, S-nitroso- N-acetyl-penicillamine (SNAP). Although NO production by iNOS did not induce myocardial injury, NO augmented release of lactate dehydrogenase from myocyte cultures after addition of H2O2(0.1 mM, 1 h). Inhibition of iNOS with Nω-nitro-l-arginine methyl ester ameliorated the effects of NO-enhancing treatments on myocardial injury and GPX activity. SNAP augmented the myocardial injury induced by H2O2. Inhibition of GPX activity with antisense oligodeoxyribonucleotide for GPX mRNA increased myocardial injury by H2O2. Results suggest that the induction of cardiac iNOS promotes myocardial injury due to oxidative stress via inactivation of the intrinsic antioxidant enzyme, GPX.

PKC mediates LPS- and phorbol-induced cardiac cell nitric oxide synthase activity and hypocontractility

1995 ◽  
Vol 269 (6) ◽  
pp. H1891-H1898 ◽  
Author(s):  
T. M. McKenna ◽  
S. Li ◽  
S. Tao
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Cardiac Myocyte ◽  
Contractile Function ◽  
Dependent Manner ◽  
Cardiac Cell ◽  
Nos Inhibitor ◽  
Oxide Synthase ◽  
Myocyte Contractility ◽  
Rat Cardiac Myocytes

Lipopolysaccharide (LPS) treatment impairs cardiac myocyte contractility in a nitric oxide synthase (NOS)-dependent manner. The objective of this study was to assess whether protein kinase C (PKC) transduces the LPS signal into an enhanced NOS activity in rat cardiac myocytes. LPS (100 ng/ml) stimulated myocyte PKC activity, inducible NOS (iNOS) expression, and NOS activity in a time- and protein synthesis-dependent fashion. Directly activating PKC with beta-phorbol 12,13-dibutyrate (beta-PDB) also induced myocyte iNOS synthesis and NOS activity and reduced electrically stimulated contractility, while the inactive alpha-PDB was ineffectual. Contractility could be restored to beta-PDB-incubated cells by superfusion with the NOS inhibitor N omega-nitro-L-arginine methyl ester. PKC blockade with sphingosine, chelerythrine, or calphostin-C precluded LPS- and beta-PDB-induced increases in NOS activity and protected contractility. Depletion of PKC by 18 h of incubation with beta-PDB in the presence of chelerythrine also blocked acquisition of enhanced NOS activity and contractile dysfunction when the myocytes were subsequently exposed to LPS. These findings suggest that PKC is a significant intracellular mediator for the effects of LPS on cardiac cell NOS activity and contractile function.

Does Nitric Oxide Have a Role in Regulating Skeletal Muscle Protein Synthesis?

1997 ◽  
Vol 92 (s36) ◽  
pp. 10P-10P
Author(s):  
M. E. Reilly ◽  
J. S. Marway ◽  
A. B. Bonner ◽  
D. L. Scott ◽  
V. R. Preedy
Keyword(s):  
Nitric Oxide ◽  
Skeletal Muscle ◽  
Protein Synthesis ◽  
Muscle Protein ◽  
Muscle Protein Synthesis ◽  
Skeletal Muscle Protein ◽  
Skeletal Muscle Protein Synthesis

scholarly journals Nitric oxide synthase acutely regulates progesterone production by in vitro cultured rabbit corpora lutea

1999 ◽  
Vol 160 (2) ◽  
pp. 275-283 ◽  
Author(s):  
A Gobbetti ◽  
C Boiti ◽  
C Canali ◽  
M Zerani
Keyword(s):  
Nitric Oxide ◽  
In Vitro Release ◽  
Inhibitory Effect ◽  
Corpora Lutea ◽  
No Donor ◽  
Nos Inhibitor ◽  
Oxide Synthase ◽  
Vitro Release

We examined the presence and the regulation of nitric oxide (NO) synthase (NOS) using in vitro cultured corpora lutea (CL) obtained from rabbits at days 4 and 9 of pseudopregnancy. The role of NO and NOS on steroidogenesis was also investigated using the same CL preparations after short-term incubations (30 min and 2 h) with the NO donor, sodium nitroprusside (NP), the NOS inhibitor, Nomega-nitro-l-arginine methyl ester (l-NAME) and prostaglandin (PG) F-2alpha. The basal NOS activity was greater in CL at day 4 than at day 9, and was also differently modulated by PGF-2alpha, depending on the age of the CL. The addition of PGF-2alpha to day 4 CL had no effect, but PGF-2alpha on day 9 caused a threefold increase in NOS activity. NP caused a two- to fivefold decrease in release of progesterone from CL of both ages, and this inhibitory effect on steroidogenesis was reversed by l-NAME. All treatments failed to modify basal androgens and 17beta-oestradiol was not detectable in either control or treated CL. These results suggest that NO is effectively involved in the regulation process of steroidogenesis, independently of 17beta-oestradiol. PGF-2alpha had no effect on day 4, but induced luteolysis on day 9, by reducing progesterone (P</=0. 01) to about 18% of control. The luteolytic action of PGF-2alpha was completely reversed by co-incubation with l-NAME, thus supporting the hypothesis that luteolysis is mediated by NO. The addition of NP or l-NAME did not modify the in vitro release of PGF-2alpha. We hypothesised that PGF-2alpha upregulates NOS activity and, consequently, the production of NO, which acutely inhibits progesterone release from day 9 CL of pseudopregnant rabbits.

Evidence that nitric oxide increases glucose transport in skeletal muscle

1997 ◽  
Vol 82 (1) ◽  
pp. 359-363 ◽  
Author(s):  
Thomas W. Balon ◽  
Jerry L. Nadler ◽  
Keyword(s):  
Nitric Oxide ◽  
Skeletal Muscle ◽  
Glucose Transport ◽  
Type I ◽  
Stimulation Protocol ◽  
No Donor ◽  
Nos Inhibitor ◽  
Exercise Induced ◽  
Oxide Synthase

Balon, Thomas W., and Jerry L. Nadler. Evidence that nitric oxide increases glucose transport in skeletal muscle. J. Appl. Physiol. 82(1): 359–363, 1997.—Nitric oxide synthase (NOS) is expressed in skeletal muscle. However, the role of nitric oxide (NO) in glucose transport in this tissue remains unclear. To determine the role of NO in modulating glucose transport, 2-deoxyglucose (2-DG) transport was measured in rat extensor digitorum longus (EDL) muscles that were exposed to either a maximally stimulating concentration of insulin or to an electrical stimulation protocol, in the presence of N G-monomethyl-l-arginine, a NOS inhibitor. In addition, EDL preparations were exposed to sodium nitroprusside (SNP), an NO donor, in the presence of submaximal and maximally stimulating concentrations of insulin. NOS inhibition reduced both basal and exercise-enhanced 2-DG transport but had no effect on insulin-stimulated 2-DG transport. Furthermore, SNP increased 2-DG transport in a dose-responsive manner. The effects of SNP and insulin on 2-DG transport were additive when insulin was present in physiological but not in pharmacological concentrations. Chronic treadmill training increased protein expression of both type I and type III NOS in soleus muscle homogenates. Our results suggest that NO may be a potential mediator of exercise-induced glucose transport.

scholarly journals Development of a gene doping detection method to detect overexpressed human follistatin using an adenovirus vector in mice

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e12285
Author(s):  
Koki Yanazawa ◽  
Takehito Sugasawa ◽  
Kai Aoki ◽  
Takuro Nakano ◽  
Yasushi Kawakami ◽  
...  
Keyword(s):  
Protein Synthesis ◽  
Sanger Sequencing ◽  
Detection Method ◽  
Muscle Protein ◽  
Muscle Protein Synthesis ◽  
Quantitative Polymerase Chain Reaction ◽  
C2c12 Cells ◽  
Detection Methods ◽  
Gene Doping ◽  
Taqman Qpcr

Background Gene doping is the misuse of genome editing and gene therapy technologies for the purpose of manipulating specific genes or gene functions in order to improve athletic performance. However, a non-invasive detection method for gene doping using recombinant adenoviral (rAdV) vectors containing human follistatin (hFST) genes (rAdV<hFST>) has not yet been developed. Therefore, the aim of this study was to develop a method to detect gene doping using rAdV<hFST>. Methods First, we generated rAdV<hFST> and evaluated the overexpression of the hFST gene, FST protein, and muscle protein synthesis signaling using cell lines. Next, rAdV<hFST> was injected intravenously or intramuscularly into mice, and whole blood was collected, and hFST and cytomegalovirus promoter (CMVp) gene fragments were detected using TaqMan-quantitative polymerase chain reaction (qPCR). Finally, to confirm the specificity of the primers and the TaqMan probes, samples from each experiment were pooled, amplified using TaqMan-qPCR, and sequenced using the Sanger sequencing. Results The expression of hFST and FST proteins and muscle protein synthesis signaling significantly increased in C2C12 cells. In long-term, transgene fragments could be detected until 4 days after intravenous injection and 3 days after intramuscular injection. Finally, the Sanger sequencing confirmed that the primers and TaqMan probe specifically amplified the gene sequence of interest. Conclusions These results indicate the possibility of detecting gene doping using rAdV<hFST> using TaqMan-qPCR in blood samples. This study may contribute to the development of detection methods for gene doping using rAdV<hFST>.

scholarly journals Effects of inhibiting nitric oxide synthase on cumulus expansion and nuclear maturation of sheep oocytes

2011 ◽  
Vol 56 (No. 6) ◽  
pp. 284-291 ◽  
Author(s):  
Heidari Amale M ◽  
Zare Shahne A ◽  
A. Abavisani ◽  
S. Nasrollahi
Keyword(s):  
Nitric Oxide ◽  
Nitric Oxide Synthase ◽  
Oocyte Maturation ◽  
In Vitro Maturation ◽  
No Donor ◽  
Cumulus Expansion ◽  
Maturation Medium ◽  
Nos Inhibitor ◽  
Oxide Synthase

Nitric oxide (NO) is a biological signaling molecule that plays a crucial role in oocyte maturation of mammalians. It is generated by the nitric oxide synthase (NOS) enzyme from l-arginine. Although the effect of NO has been shown in oocyte maturation of some species, there is no report about its effect on the in vitro maturation of sheep oocyte. So, this study aimed to investigate the importance of NO/NOS system in the in vitro maturation of ovine oocytes. Different concentrations of L-NAME (a NOS inhibitor) (0.1, 1 and 10mM) were added to maturation medium to evaluate the effect of inhibiting NOS on cumulus expansion and meiotic resumption of sheep oocytes. After 26 h culture, low and medium concentrations of L-NAME (0.1 and 1mM) had no significant effect on cumulus expansion, however, its higher concentration (10mM) decreased percentage of oocytes with total cumulus expansion as compared to control (P &lt; 0.05). The extrusion of the first polar body was also suppressed in a dose-dependent manner, so that the addition of 10mM L-NAME to maturation medium significantly stopped oocytes in GV stage (P &lt; 0.05). Moreover, to confirm the results and to evaluate if this effect is reversible, 0.1mM sodium nitroprusside (SNP, a NO donor) was added only to the maturation medium which had the highest concentration of L-NAME (10mM). The concomitant addition of NOS inhibitor with NO donor reversed the inhibitory effect of L-NAME on cumulus expansion and meiotic maturation. These results indicated that NO/NOS system is involved in the maturation of sheep oocytes.

Endogenous nitric oxide is implicated in the regulation of lipolysis through antioxidant-related effect

2000 ◽  
Vol 279 (5) ◽  
pp. C1603-C1610 ◽  
Author(s):  
Nicolas Gaudiot ◽  
Catherine Ribière ◽  
Anne-Marie Jaubert ◽  
Yves Giudicelli
Keyword(s):  
Nitric Oxide ◽  
Related Effect ◽  
No Donor ◽  
Endogenous Nitric Oxide ◽  
Dependent Protein ◽  
Diethyldithiocarbamic Acid ◽  
Isolated Adipocytes ◽  
Nos Inhibitor ◽  
Oxide Synthase ◽  
Preventive Effects

We studied the influence of nitric oxide (NO) endogenously produced by adipocytes in lipolysis regulation. Diphenyliodonium (DPI), a nitric oxide synthase (NOS) inhibitor, was found to completely suppress NO synthesis in intact adipocytes and was thus used in lipolysis experiments. DPI was found to decrease both basal and dibutyryl cAMP (DBcAMP)-stimulated lipolysis. Inhibition of DBcAMP-stimulated lipolysis by DPI was prevented by S-nitroso- N-acetyl-penicillamine (SNAP), a NO donor. This antilipolytic effect of DPI was also prevented by two antioxidants, ascorbate or diethyldithiocarbamic acid (DDC). Preincubation of isolated adipocytes with DPI (30 min) before exposure to DBcAMP almost completely abolished the stimulated lipolysis. Addition of SNAP or antioxidant during DPI preincubation restored the lipolytic response to DBcAMP, whereas no preventive effects were observed when these compounds were added simultaneously to DBcAMP. Exposure of isolated adipocytes to an extracellular generating system of oxygen species (xanthine/xanthine oxidase) or to H2O2 also resulted in an inhibition of the lipolytic response to DBcAMP. H2O2 or DPI decreased cAMP-dependent protein kinase (PKA) activation. The DPI effect on PKA activity was prevented by SNAP, ascorbate, or DDC. These results provide clear evidence that 1) the DPI antilipolytic effect is related to adipocyte NOS inhibition leading to PKA alterations, and 2) endogenous NO is required for the cAMP lipolytic process through antioxidant-related effect.

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