• Korean Journal of Plant Resources
• /
• v.31 no.4
• /
• pp.303-311
• /
• 2018

In this study, a preliminary evaluation of the antioxidant and anti-inflammatory activity of the Ficus erecta var. sieboldii (Miq.) King (FES) leaf extract has been performed to assess its potential as a natural resource for food and medicinal materials. FES was extracted using 70% EtOH and then fractionated sequentially using n-hexane, $CH_2Cl_2$, EtOAc, and n-BuOH. To screen for antioxidant and anti-inflammatory agents effectively, the inhibitory effect of the FES extracts on the production of oxidant stresses (DPPH, xanthine oxidase, and superoxide) and pro-inflammatory factors (NO, iNOS, COX-2, $PGE_2$, IL-6, and $IL-1{\beta}$) in the murine macrophage cell line RAW 264.7 activated with lipopolysaccharide (LPS) was examined. Among the sequential solvent fractions of FES, the $CH_2Cl_2$ and EtOAc fractions showed decreased production of oxidant stresses (DPPH, xanthine oxidase and superoxide), and the hexane and $CH_2Cl_2$ fractions of FES inhibited the production of pro-inflammatory factors (NO, iNOS, COX-2, and $PGE_2$). The $CH_2Cl_2$ fraction also inhibited the production of pro-inflammatory cytokines ($TNF-{\alpha}$, IL-6, and $IL-1{\beta}$). These results suggest that FES has a significant effects on the production of oxidant stresses and pro-inflammatory factors and may be used a natural resource for antioxidant and anti-inflammatory agents.

• Journal of Acupuncture Research
• /
• v.33 no.2
• /
• pp.35-49
• /
• 2016

Objectives : This study was designed to investigate the anti-osteoarthritic effects of clematidis radix pharmacopuncture on the MIA-induced osteoarthritis rats. Methods : The subjects were divided into 4 groups ; Normal rat group(N, n=5), MIA-induced osteoarthritis control rat group(C, n=5), MIA-induced osteoarthritis rat group injected with normal saline at ST36(NS, n=5), and MIA-induced osteoarthritis rat group injected with clematidis radix pharmacopuncture at ST36(CR, n=5). The experiment was conducted over a period of 21 days after injecting MIA. We analyzed body weight, hind paw weight distribution, liver and renal function, immunocytes, cykokines, inflammatory mediators, inflammatory proteins and mRNA expressions, as well as histopathological changes. Results : The CR group showed a significant increase in the hind paw weight distribution, and significant decreases in IL-$1{\beta}$, IL-6, $PGE_2$, $LTB_4$, osteocalcin, deoxypyridinoline level, the protein expression of COX2, arachidonate 5 lipoxygenase, and the mRNA expression of COX2, TNF-${\alpha}$, IL-$1{\beta}$, and NOS-II. In terms of the joint damages induced by osteoarthritis, the CR group showed a greater protective effect than group C in histopathologic observation (H&E, Safranin-O staining). Conclusion : These results demonstrated that clematidis radix pharmacopuncture had anti-inflammatory and analgesic effects. In addition, these results showed that it inhibited the progression of osteoarthritis.

• Molecules and Cells
• /
• v.21 no.3
• /
• pp.337-342
• /
• 2006

Interstitial cells of Cajal (ICCs) are pacemaker cells that activate the periodic spontaneous depolarization (pacemaker potentials) responsible for the production of slow waves in gastrointestinal smooth muscle. The effects of vasoactive intestinal polypeptide (VIP) on the pacemaker potentials in cultured ICCs from murine small intestine were investigated by whole-cell patch-clamp techniques. Addition of VIP (50 nM-$1{\mu}M$) decreased the amplitude of pacemaker potentials and depolarized resting membrane potentials. To examine the type of receptors involved in ICC, we examined the effects of the $VIP_1$ agonist and found that it had no effect on pacemaker potentials. Pretreatment with $VIP_1$ antagonist ($1{\mu}M$) for 10 min also did not block the VIP (50 nM)-induced effects. On the other hand exposure to 1H-(1,2,4)oxadiazolo(4,3-A)quinoxalin-1-one (ODQ, $100{\mu}M$), an inhibitor of guanylate cyclase, prevented VIP inhibition of pacemaker potentials. Similarly KT-5823 ($1{\mu}M$) or RP-8-CPT-cGMPS ($10{\mu}M$), inhibitors of protein kinase G (PKG) blocked the effect of VIP (50 nM) on pacemaker potentials as did N-nitro-L-arginine (L-NA, $100{\mu}M$), a non-selective nitric oxide synthase (NOS) inhibitor. These results imply that the inhibition of pacemaker activity by VIP depends on the NO-cGMP-PKG pathway.

• The Korea Journal of Herbology
• /
• v.23 no.2
• /
• pp.179-192
• /
• 2008

Objectives : Draconis Resina (DR) has been used as a traditional Korean herbal medicine since ancient times, and today it is used as a medication for wounds, tumors, diarrhea, rheumatism, in the itching of insect bites and with other conditions in the folk medicine. The aim of this study was to determine whether fractionated extracts of DR inhibit free radical generation, intracellular oxidation, production of nitrite, an index of NO, PGE2, iNOS, COX-2 and proinflammatory cytokines in lipopolysaccharide (LPS)-treated RAW 264.7 macrophages, Methods : DR extract prepared with methanol, and then fractionated with hexane, dichloromethane, ethylacetate, n-butanol and water. Inhibitory effect of DR onto free radical generation was determined by measuring DPPH, superoxide anions and nitric oxide scavenging activities in vitro. Cytotoxic activity of extracts on RAW 264.7 cells was measured using 5-(3-caroboxymeth-oxyphenyl)-2H-tetra-zolium inner salt (MTS) assay. Intracelluar oxidation was analysed by DCF-DA assay. The nitric oxide (NO) production was measured by Griess reagent system. The levels of iNOS and COX-2 expression were confirmed by western blot. And proinflammatory cytokines were measured by ELISA kit. Results : Our results indicated that fractionated extracts, especially dichloromethane and ethyl acetate extracts, significantly inhibited free radical generation, the LPS-induced H202, NO, PGE2 production and iNOS, COX-2 expression accompanied by an attenuation of TNF-${\alpha}$, IL-$1{\beta}$ and IL-6 formation in macrophages. Conclusions : Our results indicate that dichloromethane and ethyl acetate extracts of DR have potential as an agent of chronic inflammatory diseases.

• Journal of Life Science
• /
• v.27 no.1
• /
• pp.8-14
• /
• 2017

Died Gardenia jasminoides fruit is used as a dye in the food and clothes industries in Asia. The present study investigated the anti-inflammatory effects of aqueous extract of G. jasminoides fruits (GJ) in BV-2 microglial cells. GJ inhibited lipopolysaccharide-induced nitric oxide (NO) secretion, inducible nitric oxide synthase (iNOS) expression, and reactive oxygen species production, without affecting cell viability. Furthermore, GJ increased the expression of heme oxygenase-1 (HO-1) in a dose-dependent manner. Moreover, the inhibitory effect of GJ on iNOS expression was abrogated by small interfering RNA-mediated knock-down of HO-1. In addition, GJ induced nuclear translocation of nuclear factor E2-related factor 2 (Nrf2), a transcription factor that regulates HO-1 expression. GJ-mediated expression of HO-1 was suppressed by LY294002, a phosphoinositide 3-kinase (PI-3K) inhibitor, and SB203580, a p38 kinase inhibitor, but not by the extracellular signal-regulated kinase (ERK) inhibitor PD98059 or c-Jun N-terminal kinase (JNK) inhibitor SP600125. GJ also enhanced the phosphorylation of Akt and p38. These results suggest that GJ suppresses the production of NO, a pro-inflammatory mediator, by inducing HO-1 expression via PI-3K/Akt/p38 signaling. These findings illustrate a novel molecular mechanism by which extract from G. jasminoides fruits inhibits neuroinflammation.

• Biomolecules & Therapeutics
• /
• v.23 no.2
• /
• pp.180-188
• /
• 2015

This study investigated the possible effects and molecular mechanisms of diallyl disulfide (DADS) against cyclophosphamide (CP)-induced hemorrhagic cystitis (HC) in rats. Inflammation response was assessed by histopathology and serum cytokines levels. We determined the protein expressions of nuclear transcription factor kappa-B (NF-${\kappa}B$), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), and tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$), oxidative stress, urinary nitrite-nitrate, malondialdehyde (MDA), and 8-hydroxy-2'-deoxyguanosine (8-OHdG). Finally, we studied the involvement of mitogen-activated protein kinases (MAPKs) signaling in the protective effects of DADS against CP-induced HC. CP treatment caused a HC which was evidenced by an increase in histopathological changes, proinflammatory cytokines levels, urinary nitrite-nitrate level, and the protein expression of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-c-Jun N-terminal kinase (JNK), and p-extracellular signal regulated kinase (ERK). The significant decreases in glutathione content and glutathione-S-transferase and glutathione reductase activities, and the significant increase in MDA content and urinary MDA and 8-OHdG levels indicated that CP-induced bladder injury was mediated through oxidative DNA damage. In contrast, DADS pretreatment attenuated CP-induced HC, including histopathological lesion, serum cytokines levels, oxidative damage, and urinary oxidative DNA damage. DADS also caused significantly decreased the protein expressions of NF-${\kappa}B$, COX-2, iNOS, TNF-${\alpha}$, p-JNK, and p-ERK. These results indicate that DADS prevents CP-induced HC and that the protective effects of DADS may be due to its ability to regulate proinflammatory cytokines production by inhibition of NF-${\kappa}B$ and MAPKs expressions, and its potent anti-oxidative capability through reduction of oxidative DNA damage in the bladder.

• The Korean Journal of Physiology and Pharmacology
• /
• v.14 no.3
• /
• pp.177-183
• /
• 2010

Tetrahydrobiopterin (BH4), an essential cofactor for nitric oxide synthase (NOS) activity, is known to play important roles in modulating both NO and superoxide production during vascular diseases such as atherosclerosis. However, the role of BH4 in functions of vascular smooth muscle cells is not fully known. In this study, we tested the effects of BH4 and dihydrobiopterin (BH2), a BH4 precursor, on migration and proliferation in response to platelet-derived growth factor-BB (PDGF-BB) in rat aortic smooth muscle cells (RASMCs). Cell migration and proliferation were measured using a Boyden chamber and a 5-bromo-2'-deoxyuridine incorporation assay, respectively, and these results were confirmed with an ex vivo aortic sprout assay. Cell viability was examined by 2,3-bis [2-methoxy-4-nitro-5-sulfophenyl]-2H-tetrazolium-5-carboxanilide assays. BH4 and BH2 decreased PDGF-BBinduced cell migration and proliferation in a dose-dependent manner. The inhibition of cell migration and proliferation by BH4 and BH2 was not affected by pretreatment with $N^G$-nitro-L-arginine methyl ester, a NOS inhibitor. Moreover, the sprout outgrowth formation of aortic rings induced by PDGF-BB was inhibited by BH4 and BH2. Cell viability was not inhibited by BH4 and BH2 treatment. The present results suggest that BH4 and BH2 may inhibit PDGF-stimulated RASMC migration and proliferation via the NOS-independent pathway. Therefore, BH4 and its derivative could be useful for the development of a candidate molecule with an NO-independent anti-atherosclerotic function.

• The Korean Journal of Physiology and Pharmacology
• /
• v.18 no.1
• /
• pp.25-32
• /
• 2014

Nitric oxide (NO) is recognized as a mediator and regulator of inflammatory responses. NO is produced by nitric oxide synthase (NOS), and NOS is abundantly expressed in the human dental pulp cells (HDPCs). NO produced by NOS can be cytotoxic at higher concentrations to HDPCs. However, the mechanism by which this cytotoxic pathway is activated in cells exposed to NO is not known. The purpose of this study was to elucidate the NO-induced cytotoxic mechanism in HDPCs. Sodium nitroprusside (SNP), a NO donor, reduced the viability of HDPCs in a dose- and time-dependent manner. We investigated the in vitro effects of nitric oxide on apoptosis of cultured HDPCs. Cells showed typical apoptotic morphology after exposure to SNP. Besides, the number of Annexin V positive cells was increased among the SNP-treated HDPCs. SNP enhanced the production of reactive oxygen species (ROS), and N-acetylcysteine (NAC) ameliorated the decrement of cell viability induced by SNP. However, a soluble guanylate cyclase inhibitor (ODQ) did not inhibited the decrement of cell viability induced by SNP. SNP increased cytochrome c release from the mitochondria to the cytosol and the ratio of Bax/Bcl-2 expression levels. Moreover, SNP-treated HDPCs elevated activities of caspase-3 and caspase-9. While pretreatment with inhibitors of caspase (z-VAD-fmk, z-DEVD-fmk) reversed the NO-induced apoptosis of HDPCs. From these results, it can be suggested that NO induces apoptosis of HDPCs through the mitochondria-dependent pathway mediated by ROS and Bcl-2 family, but not by the cyclic GMP pathway.

• Biomedical Science Letters
• /
• v.5 no.1
• /
• pp.85-93
• /
• 1999

Nitric oxide (NO) plays an important role in immunologic defense, and influences upon the functioning of secretory tissues and cells. It also exhibits cytotoxic/cytostatic activity as one of major operating effectors of the cellular immunity system. We investigated the effects of serum on the cell damages and NO production in the mouse liver cell line BNL CL.2 to establish the role of NO. We observed that, when BNL CL.2 cells were cultured in serum-free medium, they were induced to cell damage by the stimulation of IFN-$\gamma$ alone or IFN-$\gamma$ plus LPS. Serum-starved cells showed large amount of nitrite accumulation and NO synthase (NOS) expression in response to IFN-$\gamma$ alone in dose- and time- dependent manners, but serum-supplied cells did not The production of NO was blocked by protein tyrosine kinase (PTK) inhibitors, genistein and herbimycin. These results suggest that the deprivation of serum in the BNL CL.2 cell culture medium might primed with the cells to produce NO when the cells are triggered by IFN-$\gamma$ and the involvement of PTK signal transduction pathway in the expression of NOS gene in murine hepatocytes.

• Tuberculosis and Respiratory Diseases
• /
• v.53 no.2
• /
• pp.148-160
• /
• 2002

배 경 : 쥐를 고농도의 산소에 60시간 이상 노출시켰을 때 급성 폐손상이 유발되지만 내독소를 저용량으로 투여시에는 이러한 폐손상이 경감된다고 알려져 있으나 그 기전에 대하여는 확실히 밝혀지지 않고 있다. 산화질소(nitric oxide, NO)는 내독소나 염증성 사이토카인(cytokine) 등의 자극에 의해서 폐내 여러 염증세포에서 만들어지며 이 산화질소는 경우에 따라 우리 몸에 이롭거나 해로운 양면성을 지니고 있다. 저자들은 쥐에서 고농도의 산소에 의한 폐손상이 저농도의 내독소 투여로 경감되는 기전에, 산화질소가 중요한 역할을 하는지 또는 황산화효소나 다른 항염증성 사이토카인이 중요한 역할을 하는지를 규명하고자 하였다. 방 법 : 총 120마리의 쥐 (Sprague-Dawley rat)를 24마리씩 5군으로 나누어 대조군은 실내 공기를, 고농도 산소군은 100%의 산소를 100%의 산소를 60시간 투여하였고 내독소군은 100% 산소 투여시 2일간 저용량의 내독소를 투여하였다. 다른 두 군은 산화질소 합성 억제물인 aminoguanidine(AG)과 N-nitro-L-arginine methyl ester (L-NAME)를 각각 2일간 고농도 산소와 내독소에 더하여 투여하였다. 각각의 군에서 폐손상의 정도와 사망률을 관찰하고 superoxide dismutase(SOD), catalase, nitric oxide, IL-6, IL-11을 기관지폐포세척액에서 측정하고, 고농도산소 투여군의 폐조직에서 iNOS synthase rnRNA의 발현을 비교하였다. 결 과: 1. 100%의 산소에 60시간 노출시켰을 때 쥐의 사망률은 8.3% 이었고 내독소 투여군은 4.2%, NAME 투여군이 37.5%, AG 투여군이 25%로 산화질소 합성 억제제에 의하여 사망률의 증가가 관찰되었다. 2. 폐의 손상 정도를 나타내는 폐의 wet/dry 중량비와 늑막액도 100%의 산소에 노출된 군에서 증가되었고 내독소 투여에 의하여 감소되었으며 NAME나 AG 투여군에서는 오히려 증가되었다. 3. 이러한 내독소에 의한 폐손상 억제효과가 항산화효소인 SOD나 catalase, 또는 protective cytokine인 IL-6나 IL-11등의 증가와 관련이 있는지를 관찰하였으나 이들 모두에서 유의한 변화를 관찰하지 못하였다. 4. 산화질소는 100% 산소에 노출시킨 군에서도 증가하였으나 내독소 투여군에서 유의하게 더욱 증가하였고 이는 L-NAME 나 aminoguanidine의 투여시 감소하였다. 5. iNOS mRNA의 발현도 내독소 투여군에서 유의하게 증가하였다. 결 론 : 쥐의 고농소 산소 투여에 의한 폐손상은 저용량의 내독소 투여로 경감되며, 이는 주로 내독소 투여에 의한 iNOS mRNA의 발현을 유도하여 생성된 산화질소의 증가에 기인하는 것으로 생각된다.