• 약학회지
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• 제46권5호
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• pp.343-347
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• 2002

In the present study, effects of essential oils isolated from various plants have been evaluated on lipopolysaccharide (LPS)-induced release of nitric oxide (NO), prostaglandin E$_2$(PGE$_2$) and tumor necrosis factor-a (TNF-$\alpha$) by the macrophage RAW 264.7 cells. Among the tested essential oils, essential oil of Ligularia fischeri var. spiciformis (LF-oil) significantly inhibited the LPS-induced generation of NO, PGE$_2$ and TNF-$\alpha$ in Raw 264.7 cells. Consistent with these observations, the expression of inducible NO synthase (iNOS) and cyclooxygenase (COX)-2 enzyme was inhibited by LF-oil in a concentration-dependent manner. Thus, this study suggests that inhibition of release of iNOS, COX-2 expression, and TNF-$\alpha$ by the essential oil of Ligularia fischer may be one of the mechanisms responsible for the anti-inflammatory effects of this medicinal plant.

• Natural Product Sciences
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• 제25권1호
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• pp.16-22
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• 2019

Inula helenium L. is rich source of eudesmane-type sesquiterpene lactones, mainly alantolactone and isoalantolactone, which have the various pharmacological functions. In this study, we examined the inhibitory effects of nitric oxide (NO) production of hexane, methylene chloride, ethyl acetate, butanol, and water fractions from I. helenium and investigated the anti-inflammatory effect of hexane fraction of I. helenium (HFIH) in LPS-induced RAW 264.7 cells. Quantification of alantolactone and isoalantolactone from HFIH was carried out for the standardization by multiple reaction monitoring using triple quadrupole mass spectrometer. HFIH significantly inhibited inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) protein as well as their downstream products NO and prostaglandin $E_2$ ($PGE_2$) in LPS-stimulated RAW 264.7 cells. Moreover, HFIH suppressed $NF-{\kappa}B$ transcriptional activity by decreasing the translocation of p65 to the nucleus. The in vivo study further confirmed that HFIH attenuated the paw edema induced by carrageenan in an acute inflammation model. These findings suggest that HFIH may be useful as a promising phytomedicine for inflammatory-associated diseases.

• Natural Product Sciences
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• 제25권1호
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• pp.64-71
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• 2019

The present study was conducted to investigate anti-nociceptive and anti-inflammatory effects of the leaves of Ilex latifolia Thunb (I. latifolia) in in vivo and in vitro. Writhing responses induced by acetic acid and formalin- and thermal stimuli (tail flick and hot plate tests)-induced pain responses for nociception were evaluated in mice. I. latifolia (50 - 200 mg/kg, p.o.) and ibuprofen (100 mg/kg, p.o.), a positive non-steroidal anti-inflammatory drug (NSAID), inhibited the acetic acid-induced writhing response and the second phase response (peripheral inflammatory response) in the formalin test, but did not protect against thermal nociception and the first phase response (central response) in the formalin test. These results show that I. latifolia has a significant anti-nociceptive effect that appears to be peripheral, but not central. Additionally, I. latifolia (50 and $100{\mu}g/mL$) and 3,5-di-caffeoyl quinic acid methyl ester ($5{\mu}M$) isolated from I. latifolia as an active compound significantly inhibited LPS-induced NO production and mRNA expression of the pro-inflammatory mediators, iNOS and COX-2, and the pro-inflammatory cytokines, IL-6 and $IL-1{\beta}$, in RAW 264.7 macrophages. These results suggest that I. latifolia can produce antinociceptive effects peripherally, but not centrally, via anti-inflammatory activity and supports a possible use of I. latifolia to treat pain and inflammation.

• 대한한의학방제학회지
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• 제12권2호
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• pp.163-173
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• 2004

Chelidonii Herba (CHE, Baek-gul-chae in Korean), which has its original description in Gu-Hwang-Bon-Cho, a classic book of oriental Herbal book, is widely used in the treatment of stomach cancer, jaundice, gasrtic ulcer, edema and stomach pain, in Korea, Japan and China. The present study was conducted to evaluate the effect of CHE on the nitric oxide (NO) production, iNOS and COX-2 expression in lipopolysaccharide - activated Raw 264.7 cells. After the treatment of CHE, NO production was monitored by measuring the nitrite content in culture medium, cell viability was measured by MIT assay. COX-2 and iNOS were determined by lmmunoblot analysis. The production of nitric oxide was significantly inhibited by pretreatment (1h) with CHE (0.1-0.3 mg／ml) on LPS-activated Raw264.7 cells. The expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase 2 (COX-2) protein were up-regulated by LPS, but the increased levels of iNOS and COX-2 were inhibited by pretreatment of CHE (0.1-0.3 mg／ml), respectively. Thus, the present data suggest that CHE may play an important role in adjunctive therapy in Gram-negative bacterial infections.

• 한국응용약물학회:학술대회논문집
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• 한국응용약물학회 2003년도 Annual Meeting of KSAP : International Symposium on Pharmaceutical and Biomedical Sciences on Obesity
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• pp.101-101
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• 2003

Prostaglandins (PGs) and nitric oxide (NO) produced by inducible cyclooygenase (COX-2) and nitric oxide synthase (iNOS), respectively, have been implicated as important mediators in the process of inflammation and carcinogenesis. On this line, the potential COX-2 or iNOS inhibitors have been considered as anti-inflammatory and cancer chemopreventive agents. In our continuing efforts of searching for novel cancer chemopreventive agents from natural products, we isolated natural sesquiterpenoids as potential COX-2 and iNOS inhibitors in cultured lipopolysaccharide (LPS)-activated mouse macrophage RAW 264.7 cells. Alantolactone, a natural eudesmane-type sesquiterpenoid, exhibited a potent inhibition of COX-2 (IC50 = 0.4 $\mu\textrm{g}$/$m\ell$) and iNOS activity (IC50 = 0.08 $\mu\textrm{g}$/$m\ell$) in the assay system determined by PGE2 and NO accumulation, respectively. The inhibitory potential of alantolactone on the PGE2 and NO production was well coincided with the suppression of COX-2 and iNOS protein and mRNA expression in LPS-induced macrophages. Furthermore, alantolactone inhibited NF-kB but not AP-l binding activity on nuclear extracts evoked by LPS-stimulated macrophage cells, suggesting the possible involvement of NF-kB in the regulation of COX-2 and iNOS expression. In further study with COX-2-expressing human colon HT-29 cells, alantolactone inhibited the cell proliferation, down-regulated COX-2, and inhibited the ERK phosphorylation in the early time. These results suggest that a natural sesquiterpenoid alantolactone might be a potential lead candidate for further developing COX-2 or iNOS inhibitor possessing cancer chemopreventive or anti-inflammatory activity

• 대한본초학회지
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• 제22권3호
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• pp.117-125
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• 2007

Objective : Lonicera japonica (Caprifoliaceae) has long been used for treatment of infectious diseases in oriental countries. The aim of this study was to investigative the effect by which the aqueous extract from flower of L. japonica (LJFAE) inhibited the lipopolysaccharide (LPS)-induced inflammatory mediators in murine macrophages, RAW 264.7 cells Methods : The dried flowers of L. japonica were extracted with distilled water at $100^{\circ}C$ for 7 h. The extract was filtered through 0.45 ${\mu}m$ filter, freeze-dried. The dried extract was dissolved in Hank's balanced salt solution (HBSS) and filtered through 0.22 ${\mu}m$ filter before use. Accumulated nitrite, an oxidative product of nitric oxide (NO), was measured in the culture medium by the Griess reaction. The levels of prostaglandin E2 (PGE2), tumor necrosis factor-$\alpha$ (TNF-$\alpha$), interleukin-1$\beta$ (IL-1$\beta$), and IL-6 production, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) expression were measured by enzyme-linked immunosorbent assay and Western blot analysis. Results: LJFAE (10-400 ${\mu}g$/ml) per se had no cytotoxic effect in unstimulated macrophages, but LJFAE concentration-dependently reduced NO, PGE2, TNF-, IL-l, and IL-6 production and COX-2 activity caused by stimulation of LPS. The levels of iNOS and COX-2 protein expressions were markedly suppressed by the treatment with LJFAE in a concentration dependent manner. Conclusions : These results suggest that LJFAE suppress the NO and PGE2production in macrophages by inhibiting iNOS and COX-2 expression and these properties may contribute to the anti-inflammatory activity of Lonicera japonica.

• 대한한의학회지
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• 제26권3호
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• pp.188-203
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• 2005

Objectives : This study was designed to investigate the effects of Jayun-tang extract (JYT) on the change of cerebral hemodynamics [regional cerebral blood flow (rCBF), pial arterial diameter (PAD) and mean arterial blood pressure (MABP)] in normal and cerebral ischemic rats, na to determine the mechanisms of action of JYT. Methods : We investigated whether JYT inhibits lactate dehydrogenase activity in neuronal cells and cytokines production in serum of cerebral ischemic rats. Results : 1. JYT significantly increased rCBF and PAD in a dose-dependent manner, but MABP was not changed by injecting JYT. These results suggested JYT significantly increased rCBF by dilating PAD. 2. The JYT-induced increase in rCBF was significantly inhibited from pretreatment with indomethacin (1mg/kg, i.p.), an inhibitor of cyclooxygenase and methylene blue $(10{\mu}g/kg, i.p.)$, an inhibitor of guanylate cyclase. 3. The JYT-induced dilation in PAD was significantly inhibited from pretreatment with indomethacin, but was increased by pretreatment with methylene blue. 4 The JYT-induced increase in MABP was reduced by pretreatment with indomethacin and methylene blue. 5. JYT significantly inhibited lactate dehydrogenase activity in neuronal cells. These results suggest that JYT prevented the neuronal death. 6. Both rCBF and PAD were significantly and stably increased by JYT $(10{\mu}g/kg,\;i.p.)$ during the Period or cerebral reperfusion, which contrasted with the findings of rapid and marked increase in the control group. 7. In cytokine production in the serum drawn from femoral artery 1hr after middle cerebral artery occlusion, the sample group showed significantly decreased production of $IL-1\beta$ and $TNF-\alpha$ as well as increased production of IL-10 and $TGF-\beta$ compared with rho control group. 8. In cytokine production in the serum drawn from femoral artery 1hr after reperfusion, the sample group showed significantly decreased production of $IL-1\beta$ and $TNF-\alpha$ as well as significantly increased production of IL-10 and $TGF-\beta$ compared with the control group. Conclusions : JYT mediated by cyclooxygenase had an inhibitive effect on brain damage by inhibiting lactate dehydrogenase activity, $IL-1\beta$ and $TNF-\alpha$ production, and by accelerating IL-10 and $TGF-\beta$ production. The author feels that JYT had anti-ischemic effects through the improvement of cerebral hemodynamics and inhibitive effects on brain damage.

• 대한치과교정학회지
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• 제35권5호
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• pp.351-360
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• 2005

이 연구는 $PGE_2$ 생합성 억제제인 인도메타신의 투여 시 성장기 개의 하악두 연골에 나타나는 cyclooxygenase-2 (GOX-2)와 insulin-like growth factor 1(IGF-1)의 발현과 분포를 관찰하여 인도메타신이 하악두 연골 성장에 미치는 영향을 규명하기 위하여 시행하였다 생후 12- 3주된 잡견 8마리를 4군으로 구분하였다 통상적 복용량인 인도메타신 2mg/Kg/day을 각각 7일과 14일간 투여한 군 과량인 8mg/Kg/day을 14일간 투여한 군과 무처치군인 대조군으로 구분하였으며. 하악두를 연구대상으로 하였다. 연구대상 하악두는 $5{\mu}m$ 두께의 절편을 만들어, H-E 중염색, COX-2 면역염색 IGF-1 면역염색을 시행하여 광학 현미경으로 검경하였으며, tartrate resistant acid phosphatase(TRAP) 염색 후 파연골세포의 수를 측정하여 다음과 같은 결과를 얻었다. 인도메타신은 하악두 연골의 증식대에서 COX-2와 IGF-1의 발현과 분포를 억제시켰으며, 인도메타신의 투여기간에 비례해서 파연골세포 수는 유의성 있게 감소하였다 (p<0.01) IGF-1의 발현과 분포는 인도메타신의 투여 양과 기간에 비례하여 억제되었다 이상의 결과에 의하면 인도메타신의 투여는 하악두 연골에서 COX-2와 IGF-1의 발현과 분포를 억제하고 파연골세포의 수를 감소시켜 하악두 성장을 억제할 가능성이 있음을 시사한다.

• BMB Reports
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• 제43권1호
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• pp.40-45
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• 2010

AMP-activated protein kinase (AMPK) is a heterotrimeric enzyme that plays a central role in cellular metabolic stress. Modulation of nitric oxide (NO) and cyclooxygenase-2 (COX-2) is considered a promising approach for the treatment of inflammation and neuronal diseases. In this study, the AMPK gene was fused in-frame with PEP-1 peptide in a bacterial expression vector to produce a PEP-1-AMPK fusion protein. Expressed and purified PEP-1-AMPK fusion proteins were transduced efficiently into macrophage Raw 264.7 cells in a time- and dose-dependent manner. Furthermore, transduced PEP-1-AMPK fusion protein markedly inhibited LPS-induced iNOS and COX-2 expression. These results suggest that the PEP-1-AMPK fusion protein can be used for the protein therapy of COX-2 and NO-related disorders such as inflammation and neuronal diseases.

• 분석과학
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• 제28권4호
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• pp.260-269
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• 2015

골관절염에 대한 참당귀 추출분말의 치료효과를 검토하고자 흰쥐의 monosodium iodoacetate(MIA)로 유발된 골관절염 부위에서 시료를 채취하여 염증관련 효소 및 염증성 cytokines의 발현에 대하여 참당귀 추출분말의 억제효능을 검토하였다. 고농도의 참당귀 추출분말 (50 μg/mL) 투여에서도 독성이 관찰되지 않았으며, 동일조건하에서 interleukin-1α (IL-1α)로 유발된 nitric oxide (NO)의 생성을 효과적으로 억제하였다. 특히, 관절연골 조직의 inducible nitric oxide synthase (iNOS) 및 cyclooxygenase-2(COX-2)의 발현을 농도 의존적으로 억제하였다. 따라서 참당귀 추출분말은 항염증 효능을 나타내는 농도에서 독성 없이 사용할 수 있으며, iNOS발현을 억제하여 방출되는 신호전달 물질인 NO의 생성을 억제하였다. 또한 참당귀 추출분말의 처리로 염증유발 부위에서 염증성 cytokines으로 알려진 tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β) 및 interleukin-6 (IL-6)의 발현이 억제됨을 확인하였다. 참당귀 추출분말의 항 염증효과는 TNF-α, IL-1β 및 IL-6의 혈중농도를 낮추어 염증부위뿐만 아니라 전체적으로 항염증효과를 나타내었다. 본 실험결과, 참당귀 추출분말의 투여는 MIA 또는 IL-1α로 유발되는 골관절염 동물모델에서 염증인자, 관련 효소의 발현 및 관련 신호전달 물질의 생성을 효과적으로 억제하여, 슬관절의 활액 내 glycosaminoglycan (GAG) 및 관절연골의 proteoglycan (PG)의 분해를 방지하여 골관절염의 발생을 억제할 것으로 추정되었다. 한편, 참당귀 추출분말의 주성분인 decursin은 혈중에서 2시간이내에 decursinol로 전환되어 8시간이상 LC-MS/MS로 검출되었다, 따라서 참당귀 추출분말에 의한 염증성 사이토카인 TNF-α, IL-1β 및 IL-6의 억제활성은 항염증활성이 큰 decursinol에 의한 것으로 추정된다.