cyclic-gmp and ferulic-acid

Sodium ferulate (SF) is the sodium salt of ferulic acid, which is one of the effective components of Angelica sinensis and Lignsticum chuanxiong , and plays an important role in protecting the cardiovascular system. In this study, myocardial hypertrophy was induced by angiotensin II 0.1 μmol/L in neonatal Sprague-Dawley rat ventricular myocytes. Nine groups were designed, that is, normal, normal administration, model, L-arginine (L-arg 1000 μmol/L), SF (50, 100, 200 μmol/L) group, and N G -nitro-L-arg-methyl ester 1500 μmol/L combined with SF 200 μmol/L or L-arg 1000 μmol/L group, respectively. Cardiomyocyte hypertrophy was confirmed by observing histological changes and measurements of cell diameter, protein content and atrial natriuretic factor, and β-myosin heavy chain levels of the cells. Notably, SF could inhibit significantly myocardial hypertrophy of neonatal rat cardiomyocytes in a concentration-dependent manner without producing cytotoxicity, and the levels of nitric oxide, NO synthase (NOS), endothelial NOS, and cyclic guanosine monophosphate were increased, but the level of cyclic adenosine monophosphate was decreased in cardiomyocytes. Simultaneously, levels of protein kinase C beta, Raf-1, and extracellular regulated protein kinase 1/2 (ERK1/2) were downregulated, whereas levels of mitogen-activated protein kinase phosphatase-1 were significantly upregulated. All the beneficial effects of SF were blunted by N G -nitro-L-arg-methyl ester. Overall, these findings reveal that SF can inhibit angiotensin II-induced myocardial hypertrophy of neonatal rat cardiomyocytes, which is closely related to activation of endothelial NOS/NO/cyclic guanosine monophosphate, and inhibition of protein kinase C and mitogen-activated protein kinase signaling pathways.

Topics: Angiotensin II; Animals; Cardiomegaly; Coumaric Acids; Cyclic GMP; Esters; Guanosine Monophosphate; Myocytes, Cardiac; Nitric Oxide; Nitric Oxide Synthase Type III; Rats; Rats, Sprague-Dawley; Signal Transduction

Ferulic acid (FA) is a polyphenol pertaining to the class of hydroxycinnamic acids present in numerous foods of a plant origin. Its dietary consumption leads to the formation of several phase I and II metabolites in vivo, which represent the largest amount of ferulates in the circulation and in the intestine in comparison with FA itself. In this work, we evaluated their efficacy against the proinflammatory effects induced by lipopolysaccharide (LPS) in intestinal Caco-2 cell monolayers, as well as the mechanisms underlying their protective action. LPS-induced overexpression of proinflammatory enzymes such as inducible nitric oxide synthase (iNOS) and the consequent hyperproduction of nitric oxide (NO) and cyclic guanosine monophosphate (cGMP) were limited by physiological relevant concentrations (1 µM) of FA, its derivatives isoferulic acid (IFA) and dihydroferulic acid (DHFA), and their glucuronidated and sulfated metabolites, which acted upstream by limiting the activation of MAPK p38 and ERK and of Akt kinase, thus decreasing the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-ĸB) translocation into the nucleus. Furthermore, the compounds were found to promote the expression of Nrf2, which may have contributed to the downregulation of NF-ĸB activity. The overall data show that phase I/II metabolites retain the efficacy of their dietary free form in contrasting inflammatory response.

Topics: Caco-2 Cells; Cell Survival; Coumaric Acids; Cyclic GMP; Enterocytes; Extracellular Signal-Regulated MAP Kinases; Gene Expression Regulation; Humans; I-kappa B Proteins; Inflammation; Lipopolysaccharides; NF-E2-Related Factor 2; Nitric Oxide; Nitric Oxide Synthase Type II; p38 Mitogen-Activated Protein Kinases; Proto-Oncogene Proteins c-akt

Ferulic acid (FA) is an active component of the traditional Chinese herb Angelica sinensis. Numerous health benefits have been attributed to FA, but few studies have investigated the effects of FA on osteoblasts (Obs). Our work studied the effects of FA on proliferation, differentiation, and mineralization of rat calvarial Obs and examined the signaling pathways involved. Cell proliferation and differentiation were evaluated by Cell Counting Kit-8 (CCK-8) and alkaline phosphatase (ALP) assay kit, respectively. Cyclic guanosine monophosphate (cGMP)-dependent protein kinase II (PKGII) expression was silenced by small interfering RNA (siRNA). The mRNA expression was investigated by semi-quantitative PCR. FA (40-2560 μM) promoted Ob proliferation and differentiation; at 40-640 μM, FA stimulated calcified nodule formation and increased the expression of osteogenic genes encoding osteopontin and collagen-l. FA (40-2560 μM) increased cGMP levels in Obs and upregulated the expression of PKGII, EnaCα, and ENaCγ mRNAs. Downregulated ENaCα mRNA expression in Obs transfected with the siRNA for PKGII was reversed when FA was introduced into Obs. These results demonstrated that FA promoted proliferation, differentiation, and mineralization of Obs in vitro, and enhanced osteogenic genes expression partly through the cGMP-PKGII-ENaC signaling pathway.

Topics: Alkaline Phosphatase; Animals; Cell Differentiation; Cell Proliferation; Coumaric Acids; Cyclic GMP; Molecular Structure; Osteoblasts; Osteogenesis; Osteopontin; Rats; RNA, Messenger; RNA, Small Interfering; Signal Transduction

Ferulic acid (FA) produces protective effects against cardiovascular dysfunctions. However, the mechanisms of FA is still not known. Here we examined the antithrombotic effects of FA and its potential mechanisms. Anticoagulation assays and platelet aggregation was evaluated in vitro and in vivo. Thromboxane B2 (TXB2), cyclic adenosine monophosphate(cAMP), and cyclic guanosine monophosphate (cGMP) was determined using enzyme immunoassay kits. Nitric oxide (NO) production was measured using the Griess reaction. Protein expression was detected by Western blotting analysis. Oral administration of FA prevented death caused by pulmonary thrombosis and prolonged the tail bleeding and clotting time in mice,while, it did not alter the coagulation parameters, including the activated partial thromboplastin time (APTT), prothrombin time (PT), and thrombin time (TT). In addition, FA (50-200 µM) dose-dependently inhibited platelet aggregation induced by various platelet agonists, including adenosine diphosphate (ADP), thrombin, collagen, arachidonic acid (AA), and U46619. Further, FA attenuated intracellular Ca(2)(+) mobilization and TXB2 production induced by the platelet agonists. FA increased the levels of cAMP and cGMP and phosphorylated vasodilator-stimulated phosphoprotein (VASP) while decreased phospho-MAPK (mitogen-activated protein kinase) and phosphodiesterase (PDE) in washed rat platelets, VASP is a substrate of cyclic nucleotide and PDE is an enzyme family responsible for hydrolysis of cAMP/cGMP. These results suggest that antithrombotic activities of FA may be regulated by inhibition of platelet aggregation, rather than through inhibiting the release of thromboplastin or formation of thrombin. The mechanism of this action may involve activation of cAMP and cGMP signaling.

Topics: Animals; Blood Coagulation; Calcium; Cell Adhesion Molecules; Coumaric Acids; Cyclic AMP; Cyclic GMP; Fibrinolytic Agents; Hemorrhage; Intracellular Space; Male; Mice; Microfilament Proteins; Mitogen-Activated Protein Kinases; Nitric Oxide; Phosphoproteins; Phosphoric Diester Hydrolases; Phosphorylation; Rats; Signal Transduction; Superoxides; Thrombosis; Thromboxane B2

Shunaoxin pill (SNX), one of the famous classical recipes in traditional Chinese medicine, is developed from the "Decoction of Xionggui". It has been used for treatment of cerebrovascular related diseases. It is well known that vasodilatation plays a very important role in cerebrovascular diseases. The effect of SNX on vasorelaxant activity has not yet been explored. Therefore, we aimed to investigate the vasorelaxant effects of SNX on isolated rat thoracic aorta so as to assess some of the possible mechanisms. We also investigate the gasotransmitter signaling pathway involved which has been rarely reported in isolated rat thoracic aorta before.. The present study was performed to examine the vasodilative activity of SNX and its mechanisms in isolated rat thoracic aorta.. SNX was studied on isolated rat thoracic aorta in vitro, including endothelium-intact and endothelium-denuded aortic rings. In present study, specific inhibitors including soluble guanylate cyclase (sGC) inhibitor 1 H-[1,2,4]oxadiazolo[4,3-a]quinoxaline-1-one (ODQ), cyclooxygenase (COX) inhibitor indomethacin (INDO), NO synthase inhibitor NG-nitro-l-arginine methyl ester (L-NAME), heme oxygenase-1 (HO-1) inhibitor zinc-protoporphyrin (ZnPP), cystathionine γ-lyase (CSE) inhibitor DL-Propargylglycine (PAG), non-selective K(+) channel inhibitor tetraethylammonium chloride (TEA), KV channel inhibitor 4-Aminopyridine (4-AP), and KATP channel inhibitor Glibenclamide (Gli) were used, they were added 20min before NE contraction and then added SNX to induce vasodilation.. Removal of endothelium or pretreatment of aortic rings (intact endothelium) with L-NAME, ODQ or ZnPP significantly blocked SNX-induced relaxation. Pretreatment with the non-selective K(+) channel inhibitor TEA, KV channel inhibitor 4-AP or the KATP channel inhibitor Gli, none of them had influences on the SNX-induced response (p>0.05). Besides, SNX inhibited the contraction triggered by NE in endothelium-denuded rings in Ca(2+)-free medium. SNX also produced rightward parallel displacement of CaCl2 curves.. These results suggest that SNX can induce less endothelium-dependent and more endothelium-independent vascular relaxation. The NO/cGMP and HO/CO pathways, blockade of Ca(2+) channels are inhibition of IP3R mediated Ca(2+) mobilization from intracellular stores, are likely involved in this relaxation. Furthermore, the underlying mechanisms of combined compositions in SNX await further investigations.

Topics: 4-Butyrolactone; Animals; Aorta, Thoracic; Calcium; Calcium Channel Blockers; Calcium Channels; Carbon Monoxide; Coumaric Acids; Cyclic GMP; Drugs, Chinese Herbal; Heme Oxygenase (Decyclizing); In Vitro Techniques; Male; Nitric Oxide; Norepinephrine; Potassium Chloride; Rats, Wistar; Tablets; Vasodilator Agents

Development of intracellular calcium overload is an important pathophysiological factor in myocardial ischemia/reperfusion or anoxia/reoxygenation injury. Recent studies have shown that Sodium Ferulate (SF) stimulates nitric oxide (NO) production and exerts a cardioprotective effect in the ischemia-reperfused heart. However, it has not been determined whether the cardioprotection of SF is associated with suppression of Ca(2+) overload via NO/cyclic GMP (cGMP)/cGMP-dependent protein kinase (PKG) pathway. In this work, after cardiomyocytes were incubated with 100, 200, 400, or 800 microM SF for 3 h, anoxia/reoxygenation injury was induced and intracellular Ca(2+) concentration, NO synthase (NOS) activity, guanylate cyclase activity, NO, and cGMP formation were measured appropriately. The results showed that treatment with SF concentration-dependently inhibited calcium overload induced by anoxia/reoxygenation. We also demonstrated that SF (100-800 microM) concentration dependently enhanced NO and cGMP formation through increasing NOS activity and guanylate cyclase activity in the cardiomyocytes. On the contrary, inhibition of calcium overload by SF was markedly attenuated by addition of an NOS inhibitor, an NO scavenger, an soluble guanylate cyclase inhibitor, and a PKG inhibitor: N(G)-nitro-l-arginine methyl ester (L-NAME, 100 microM), 2-(4-carboxyphenyl)-4,4,5,5-tetramethylimidazole-1-oxyl-3-oxide (c-PTIO, 1.0 microM), 1H-[1, 2, 4] oxadiazolo [4, 3-alpha] quinoxalin-1-one (ODQ, 20 microM) and KT5823 (0.2 microM), respectively. Our findings indicate that SF significantly attenuates anoxia/reoxygenation-induced Ca(2+) overload and improves cell survival in cultured cardiomyocytes through NO/cGMP/PKG signal pathway.

Topics: Animals; Animals, Newborn; Calcium; Coumaric Acids; Cyclic GMP; Cyclic GMP-Dependent Protein Kinases; Gene Expression Regulation; Guanylate Cyclase; Hypoxia; Myocytes, Cardiac; Nitric Oxide; Nitric Oxide Synthase; Oxygen; Rats; Rats, Sprague-Dawley; Reperfusion Injury; Signal Transduction; Solubility

Ferulic acid (FA), a phytochemical constituent, has antihypertensive effects, but a detailed understanding of its effects on vascular function remains unclear. The vasoreactivity of FA was assessed using aortic rings isolated from normotensive Wistar Kyoto (WKY) and spontaneously hypertensive rats (SHR).. The effects of FA (10(-5) to 10(-3) mol/L) on vasodilatory responses were evaluated based on contractile responses induced by phenylephrine (10(-6) mol/L) in thoracic aortic rings from male WKY rats and SHR. Basal nitric oxide (NO) bioavailability in the aorta was determined from the contractile response induced by the NO synthase inhibitor N(G)-nitro-L-arginine methyl ester (L-NAME, 10(-4) mol/L). The effects of FA on the production of NADPH-dependent superoxide anion were examined in SHR aortas. The impact of hydroxyhydroquinone, a generator of superoxide anions, on the FA-induced enhancement in acetylcholine-stimulated vasodilation was also investigated.. The FA (10(-3) mol/L)-induced relaxation was partially blocked by removal of the endothelium or by pretreating SHR aortas with L-NAME. FA increased NO bioavailability, and decreased NADPH-dependent superoxide anion levels in SHR aortas. Ferulic acid improved acetylcholine-induced endothelium-dependent vasodilation in SHR, but not in WKY. Furthermore, the simultaneous addition of hydroxyhydroquinone significantly inhibited the increase in acetylcholine-induced vasodilation by FA.. Ferulic acid restores endothelial function through enhancing the bioavailability of basal and stimulated NO in SHR aortas. The results explain, in part, the mechanisms underlying the effects of FA on blood pressure (BP) in SHR.

Topics: Acetylcholine; Animals; Antihypertensive Agents; Aorta; Coumaric Acids; Cyclic GMP; Endothelium, Vascular; Hydroquinones; Hypertension; Male; NADP; NG-Nitroarginine Methyl Ester; Rats; Rats, Inbred SHR; Rats, Inbred WKY; Superoxides; Thromboxane B2; Vasodilation

To investigate the cardioprotective effects of sodium ferulate (SF) mediated by nitric oxide on ischemia-reperfusion injured myocardium.. Fifty-six SD rats were killed. Their hearts were isolated and randomly divided into 7 equal groups: ischemia/reperfusion group (I/R group), ischemic/preconditioning (IP group, to be perfumed with anoxic and reoxygenated fluid several times so as to produce protection against continuous and severe ischemia), SF pretreatment group (to be perfused with fluid with SF and then made I/R model), L-NAME + SF group (to be perfused with fluid with SF and L-NAME, a NO inhibitor, and then made I/R model), glibenclamide (Glib) + SF group (to be perfused with Glib + SF and then made I/R model)), L-NAME + Glib + SF group (to be perfused with L-NAME + Glib + SF and then made I/R model)), and control group (normal isolated hearts). Electrocardiography was conducted and left ventricular systolic pressure (LVSP), dp/dt(max), and heart rate (HR) were measured. By the end of the experiment 0.5 grams of tissue was taken from the left ventricle. The levels of superoxide dismutase (SOD), malonyldialdehyde (MOD), NO, and cyclic guanosine monophosphate (cGMP) were detected.. The levels of LVSP, dp/dt(max), and HR of the SF pretreatment group and IP group were all significantly higher than those of the I/R group (all P < 0.01) without significant differences between these 2 groups (all P > 0.05). The levels of LVSP, dp/dt(max), and HR of the L-NAME, Glib + SF, and L-NAME + Glib + SF groups were all significantly lower than those of the SF pretreatment group and IP group (all P < 0.01). Ventricular extrasystole (VE) and ventricular tachycardia (VT) during re-perfusion period occurred in all hearts of the I/R group, however, the incidence rates of VE and VT of the IP and SF groups were all significantly lower than those of the I/R group (all P < 0.01), however, without significant differences between the IP and SF groups (all P > 0.05). The incidence rate of VE and VT of the L-NAME, Glib + SF, and L-NAME + Glib + SF groups were all significantly higher than those of the SF group (all P < 0.01). The myocardium MDA content of the I/R group was significantly higher and the SOD activity significantly lower in comparison with the control group (both P < 0.01); the myocardium MDA contents of the IP and SF groups were significantly lower and the SOD activity levels significantly higher in comparison with the I/R group (all P < 0.01), however, with significant differences between theses 2 groups (both P > 0.05); the myocardium MDA contents were significantly higher and the SOD activity levels significantly lower in the L-NAME, Glib + SF, and L-NAME + Glib + SF groups in comparison with the SF and IP groups (all P < 0.01). The myocardium NO(2)(-)/NO(3)(-) and cGMP contents of the I/R group were both significantly lower than those of the control group (both P < 0.01), and the myocardium NO(2)(-)/NO(3)(-) and cGMP contents of the IP and SF groups were both significantly higher than those of the I/R group (both P < 0.01), however, without significant differences between these 2 groups (both P > 0.05). The myocardium NO(2)(-)/NO(3)(-) and cGMP contents of the L-NAME, Glib + SF, and L-NAME + Glib + SF groups were all significantly lower than those of the SF group (all P < 0.01).. SF pretreatment significantly improves the releasing of NO, decreases oxygen free radicals, and relieves myocardial ischemia reperfusion injury. The opening of the ATP-sensitive potassium channels induced by the cGMP way of NO activation may be an important pathway in the cardioprotective effects of SF pretreatment.

Topics: Animals; Cardiotonic Agents; Coumaric Acids; Cyclic GMP; Enzyme Inhibitors; Female; In Vitro Techniques; Ischemic Preconditioning, Myocardial; Male; Myocardial Reperfusion Injury; NG-Nitroarginine Methyl Ester; Nitric Oxide; Nitric Oxide Synthase; Perfusion; Random Allocation; Rats; Rats, Sprague-Dawley

The capacity of human sperm fertilization principally depends on sperm motility and membrane integrity. Reactive oxygen species, such as superoxide anion and hydrogen peroxide, are known to impair sperm motility and membrane integrity by inducing membrane lipid peroxidation (LPO). Ferulic acid (FA), an effective constituent in various medicinal herbs, has recently been shown to scavenge oxygen free radicals and increase the intracellular cAMP and cGMP. The aim of this study is to investigate the effects of FA on human sperm motility, viability, lipid peroxidation, and cyclic nucleotides in fertile and asthenozoospermic infertile individuals in vitro. The sperm samples were obtained from 10 fertile volunteers and 10 asthenozoospermic infertile patients. Washed spermatozoa were incubated at 37 degrees C in Ham's F-10 medium with 0, 0.1, 0.2, 0.4, 0.8, or 1.6 mM of FA. Samples were analyzed for viability, determined by eosin-Y dye exclusion method at 0, 1, 2, 3, 5, and 6 h of incubation; motility, determined by the trans-membrane migration method within 2 h of incubation; LPO, determined by thiobarbituric acid (TBA) method at 3 h of incubation and the intracellular cAMP and cGMP, determined, respectively, by 3H-cAMP and 125I-cGMP radioimmunoassay at 3 h of incubation. The results showed: in both fertile and infertile spermatozoa, the viability, trans-membrane migration ratio (TMMR) and the levels of intracellular cAMP and cGMP in FA-treated spermatozoa were significantly higher than those of spermatozoa in control groups, while TBA-reactive substances contents in treated spermatozoa were significantly lower than those in control spermatozoa. The effects of FA on these processes were concentration dependent. These data suggested that FA is beneficial to sperm viability and motility in both fertile and infertile individuals, and that reduction of lipid peroxidative damage to sperm membranes and increase of intracellular cAMP and cGMP may be involved in these benefits. It is possible that FA may be used for cure of asthenozoospermic infertility.

Topics: Adult; Cell Survival; Coumaric Acids; Cyclic AMP; Cyclic GMP; Fertility; Free Radical Scavengers; Humans; In Vitro Techniques; Infertility, Male; Lipid Peroxidation; Male; Reactive Oxygen Species; Sperm Motility; Spermatozoa