• Journal of Life Science
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• v.21 no.12
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• pp.1689-1697
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• 2011

In this study, we investigated the anti-inflammation effect of Persicaria thunbergii (P. thunbergii) on RAW 264.7 murine macrophage cells. The anti-inflammatory activity of P. thunbergii was determined by measuring expression of the LPS-induced inflammatory proteins, inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2) and nuclear factor-${\kappa}B$ (NF-${\kappa}B$), and the production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$). Methanol extract of P. thunbergii decreased the expression of iNOS, COX-2 and NF-${\kappa}B$, and increased the expression of HO-1 in LPS-stimulated RAW264.7 cells. Methanol extract was fractioned by n-butanol, hexane and ethyl acetate (EtOAc) and each fraction was tested for inhibitory effects on inflammation. Among the sequential solvent fractions, the EtOAc soluble fraction was investigated by the expression of prostaglandin $E_2$ ($PGE_2$), and showed decreasing form to the dose-dependent manner. EtOAc extract showed the most effective inhibitory activity of the expression of iNOS, COX-2 and NF-${\kappa}B$, and the production of NO. The study showed that P. thunbergii has anti-inflammatory activity through the decrease of NO and inhibition of iNOS, COX-2, $PGE_2$ and NF-${\kappa}B$ expression, and by the increase of HO-1 enzyme. This study needs for more investigation to find out the most effective single compound with anti-inflammatory activity.

• Journal of Acupuncture Research
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• v.23 no.5
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• pp.199-206
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• 2006

Objectives : The aim of this study was to investigate the effect of various electroacupuncture stimulation on neuronal nitric oxide synthase(nNOS) in cerebral cortex, brain stem, cerebellum of spontaneously hypertensive rats. Methods : We evaluated the changes of nNOS-positive neurons using a immunohistochemical method. The staining intensity of nNOS positive neurons was assessed in a quantitative fashion using a microdensitometrical method based on optical density by means of an image analyzer. Results : The average optical density of nNOS-positive neurons of 100 Hz (bipolar square wave 0.2 ms duration and 100 Hz frequency) electroacupuncture treatment group significantly decreased in most cortical areas comparison between the manual acupuncture and 2 Hz (bipolar square wave 0.2 ms duration and 2 Hz frequency) electroacupuncture groups. In the brain stem, the optical density of nNOS-positive neuron at superficial gray layer of the superior colliculus area, dorsolateral periaqueductal gray area and paralemniscal nucleus were same as cerebral cortex. Conclusion : We conclude that the morphological evidence for nNOS-positive neurons may be have regional change in cerebral cortex brain stem and cerebellum according to various electroacupuncture stimulations.

• Journal of Nutrition and Health
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• v.50 no.3
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• pp.236-245
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• 2017

Purpose: The aim of this study was to assess the effects of guava leaf extract (GLE) supplementation on antioxidant enzyme activity and expression of endothelial nitric oxide synthase (eNOS) mRNA in ovariectomized rats. Methods: All animals were randomly assigned to four groups (n = 7 for each group): non-ovariectomized control (Sham), the ovariectomized control (OVX), ovariectomy + 150 mg/kg b.w. of GLE ($OVX{\cdot}GL$), and ovariectomy + 300 mg/kg b.w. of GLE ($OVX{\cdot}GH$). Treatment groups were administered GLE for 8 weeks every day. Results: Body weight gain was significantly reduced in the $OVX{\cdot}GL$ group compared with the OVX group (p < 0.05). The level of serum $17{\beta}$-estradiol (E2) was significantly lower in the OVX groups than the Sham group (p < 0.05). Serum triglyceride (TG) and HDL-cholesterol levels were not significantly different between all groups. However, serum total cholesterol (TC) level was significantly reduced in the $OVX{\cdot}GH$ group compared with the OVX group (p < 0.05). Serum free fatty acid (FFA) level and liver TG level were significantly reduced in both $OVX{\cdot}GL$ and $OVX{\cdot}GH$ groups compared with the OVX group (p < 0.05). Furthermore, serum glutathione peroxidase (GPx) activity was significantly elevated in the GLE groups (p < 0.05). The mRNA expression level of GPx was not affected by ovariectomy. However, administration of GLE resulted in significantly increased nuclear factor erythroid 2-related factor (Nrf2) and catalase (CAT) mRNA expression levels in the liver (p < 0.05). In addition, liver nitric oxide (NO) level was significantly reduced in the $OVX{\cdot}GH$ group compared with the OVX group (p < 0.05). Expression level of endothelial nitric oxide synthase (eNOS) was significantly elevated in the $OVX{\cdot}GH$ group compared with the OVX group (p < 0.05). Conclusion: These results suggest that GLE could have protective effects in OVX rats by stimulating eNOS expression and improving the antioxidant defense system.

• Journal of Ginseng Research
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• v.23 no.2 s.54
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• pp.81-87
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• 1999

The effect of saponin and non-saponin of Panax Red Ginseng on the blood pressure and nitric oxide production were investigated in the conscious free moving one-kidney, one-clip Goldbaltt hypertensive (lK, 1C-GBH) rats. Mean blood pressure in the control and lK, 1C-GBH rats was decreased by the administration of ginseng saponin (100 mg/kg, i.v.). The hypotensive effect induced by ginseng saponin was reached maximum at 2-4 minutes and was slowly recovered to the initial level of blood pressure. Also ginseng saponin induced reflex tachycardia in the conscious both rats. Contrast to the response induced by ginseng saponin, hypotensive effect induced by non-saponin of panax ginseng is minimal. Plasma nitric oxide concentration was increased by the treatment of ginseng saponin (100 mg/kg, i.p for 5 days) in both rats. It has been shown by western blotting that the expression level of the protein for endothelial nitric oxide synthase (eNOS) in the aorta of rats was not increased by the treatment of ginseng saponin (100 mg/kg, i.p). However, eNOS activity in aortic homogenates of both rats were increased by the treatment of ginseng saponins. From the above results, the hypotensive effect of saponin was greater than that of non-saponin of Panax Red Ginseng. The lowering effect of blood pressure by ginseng saponin may be due to the increase of plasma nitric oxide concentration via the increase of endothelial nitric oxide synthase activity in the renovascular hypertensive and control rats.

• Preventive Nutrition and Food Science
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• v.15 no.1
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• pp.14-18
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• 2010

Stevia rebaudiana (SR) is an herb used traditionally as a sweetener in Paraguay and Brazil, whose use is spreading to other countries, such as Japan, Korea and China. In addition to its low calorie sweet taste, SR appears to have other beneficial properties, such as hypotensive capabilities and inflammation reduction. To identify the bioactive natural constituents exerting anti-inflammatory activities, we examined the EtOAc fraction of SR. In the inflammatory mediator inhibitory assay from lipopolysaccharide (LPS)-activated macrophages, the EtOAc fraction significantly, and dose dependently, inhibited the enhanced production of nitric oxide (NO) and inducible nitric oxide synthase (iNOS) expression. We also found that treatment of cells with the EtOAc fraction significantly inhibited LPS-stimulated nuclear factor-${\kappa}B$ (NF-${\kappa}B$) reporter gene expression. Such inhibition of NF-${\kappa}B$ was closely associated with the inhibition of interleukin-6 (IL-6) and the monocyte chemoattractant protein-1 (MCP-1). Therefore, we suggest that SR has the potential for development as a functional food for the treatment of immune diseases, such as rheumatoid arthritis and lupus.

• The Korean Journal of Physiology and Pharmacology
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• v.3 no.4
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• pp.375-382
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• 1999

Growing evidence indicates that enhanced generation or actions of nitric oxide (NO) are implicated in the pathogenesis of hypertension in spontaneously hypertensive rats and diabetic nephropathy in streptozotocin (STZ)-induced diabetic rats. We investigated whether inducible nitric oxide synthase (iNOS) expression in STZ-induced diabetic rats is responsible for the alterations of vascular reactivity. Diabetic state was confirmed 28 days after injection of STZ (i.p) in rats by measuring blood glucose. In order to evaluate whether short term (4 weeks) diabetic state is related with altered vascular reactivity caused by iNOS expression, isometric tension experiments were performed. In addition, plasma nitrite/nitrate (NOx) levels and expression of iNOS in the lung and aorta of control and STZ-treated rats were compared by using Griess reagent and Western analysis, respectively. Results indicated that STZ-treated rats increased the maximal contractile response of the aorta to phenylephrine (PE), and high $K^+,$ while the sensitivity remained unaltered. Endothelium-dependent relaxation, but not SNP-mediated relaxation, was reduced in STZ-treated rats. Plasma nitrite/nitrates are significantly increased in STZ-treated rats compared to controls. The malondialdehyde (MDA) contents of liver, serum, and aorta of diabetic rats were also significantly increased. Furthermore, nitrotyrosine, a specific foot print of peroxynitrite, was significantly increased in endothelial cells and smooth muscle layers in STZ-induced diabetic aorta. Taken together, the present findings indicate that enhanced release of NO by iNOS along with increased lipid peroxidation in diabetic conditions may be responsible, at least in part, for the augmented contractility, possibly through the modification of endothelial integrity or ecNOS activity of endothelium in STZ-diabetic rat aorta.

• Korean Journal of Medicinal Crop Science
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• v.16 no.5
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• pp.326-331
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• 2008

To search for immunoactive natural products exerting anti-inflammatory activity, we have evaluated the effects on the water extracts of Artemisia princeps Pampanini (APP) on lipopolysaccharide-induced nitric oxide (NO), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), and prostaglandin $E_2$ ($PGE_2$) production by RAW 264.7 macrophage cell line. Our data indicate that this extract is a potent inhibitor of NO production and it also significantly decreased PGE2 and TNF-$\alpha$ production. Consistent with these results, the protein and mRNA expression level of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was inhibited by water extracts of APP in a dose-dependent manner. These results suggest that APP may exert anti-inflammatory and analgesic effects possibly by suppressing the inducible NO synthase and COX-2 expressions.

• Advances in Traditional Medicine
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• v.10 no.3
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• pp.155-164
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• 2010

Lycii fructus is the fruit of Lycium chinense Miller and is part of the Solanaceae family. Lycii fructus produces various effects such as hypotensive, hypoglycemic, anti-pyretic, and anti-stress activities. Lycii fructus is known to contain betaine, carotene, nicotinic acid, zeaxanthin, and cerebroside. In the present study, the effects of Lycii fructus aqueous extract on lipopolysaccharide (LPS)-induced inflammation in murine raw 264.7 macrophage cells were investigated. In this study we utilized the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, reverse transcriptionpolymerase chain reaction (RT-PCR), Western blotting, and nitric oxide (NO) detection. Lycii fructus aqueous extract suppressed NO production by inhibiting the LPS-induced expressions of inducible nitric oxide synthase (iNOS) and tumor necrosis factor-alpha (TNF-$\alpha$) mRNA and iNOS protein in murine raw 264.7 macrophage cells. Also, Lycii fructus aqueous extract suppressed the activation of nuclear factor-kappa B (NF-${\kappa}B$) in the nucleus. These results demonstrated that Lycii fructus aqueous extract causes an anti-inflammatory effect that was likely produced by the suppression of iNOS expression through the down-regulation of NF-$\hat{e}B$ binding activity.

• The Korean Journal of Physiology and Pharmacology
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• v.20 no.5
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• pp.539-545
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• 2016

Nafamostat mesilate (NM), a synthetic serine protease inhibitor, has anticoagulant and anti-inflammatory properties. The intracellular mediator and external anti-inflammatory external signal in the vascular wall have been reported to protect endothelial cells, in part due to nitric oxide (NO) production. This study was designed to examine whether NM exhibit endothelium dependent vascular relaxation through Akt/endothelial nitric oxide synthase (eNOS) activation and generation of NO. NM enhanced Akt/eNOS phosphorylation and NO production in a dose- and time-dependent manner in human umbilical vein endothelial cells (HUVECs) and aorta tissues obtained from rats treated with various concentrations of NM. NM concomitantly decreased arginase activity, which could increase the available arginine substrate for NO production. Moreover, we investigated whether NM increased NO bioavailability and decreased aortic relaxation response to an eNOS inhibitor in the aorta. These results suggest that NM increases NO generation via the Akt/eNOS signaling pathway, leading to endothelium-dependent vascular relaxation. Therefore, the vasorelaxing action of NM may contribute to the regulation of cardiovascular function.

• Herbal Formula Science
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• v.20 no.2
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• pp.65-82
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• 2012

Objectives : The aim of this study was verification of the anti-oxidative and anti-inflammatory effects of Odukhwan(ODH) and Sasinhwan(SSH) in mouse macrophage, RAW264.7 cells. Methods : To investigate the anti-oxidative effect and scavenging activities of DPPH radical, superoxide anions, nitric oxide and peroxynitrite were measured. Cytotoxic activity of extract of ODH and SSH on RAW264.7 cells was measured using MTS assay. To proof the reductive activity of intracellular oxidation, DCFH-DA assay was performed. The nitric oxide(NO) production was measured and pro-inflammatory cytokines and $PGE_2$ were measured by ELISA kit. The levels of inducible nitric oxide synthase(iNOS), cyclooxygenase-2(COX-2) and nuclear NF-${\kappa}B$ p65 expression were detected by western blot. Results : After those analyses, we bring to a conclusion as follows. Both herbal formulations scavenged DPPH radical and nitric oxide. But ODH had no scavenging activity of superoxide anions and SSH had low scavenging activity in peroxynitrite. And the results indicated that ODH and SSH inhibited the LPS-induced NO, $PGE_2$ production and iNOS, COX-2 expression accompanied by an attenuation of IL-$1{\beta}$ and IL-6 production in RAW264.7 cells. They also have suppression effects of LPS-induced NF-${\kappa}B$ activation. Conclusions : ODH and SSH have anti-oxidative and anti-inflammatory effects and they may be a part of database for development of new anti-oxidative and anti-inflammatory drugs.