• Journal of Chest Surgery
• /
• 제32권2호
• /
• pp.138-143
• /
• 1999

배경: 혈관 내피층에서 분비되어 평활근층 이완을 일으키는 물질의 본체는 산화질소(nitric oxide, NO)이며 NO synthase(NOS)에는 뇌형(brain NOS, bNOS), 내피세포형(endothelial constitutive NOS, ecNOS) 및 유도형 (inducible NOS, iNOS) 등 세 가지 동위효소가 있음이 알려져 있다. 고혈압은 혈관 내피층 기능 이상을 보임이 알려져 있으나 NOS 동위 효소의 변화를 포함한 세포내 기전은 아직 확실치 않다. 저자들은 고혈압 기전을 구명하기 위한 일환으로 고혈압 혈관에서 NOS 동위효소가 어떻게 변화되는가 조사하고자 하였다. 대상 및 방법: 흰쥐에서 two-kidney, one clip (2K1C) 고혈압과 deoxycorticosterone acetate(DOCA)-salt 고혈압을 일으켰다. 4주 뒤 고혈압이 일어난 것을 확인하고 적출 흉부 대동맥 표본에서 Western blot 분석에 의한 NOS 동위효소 발현 조사 및 비색법에 의한 조직내 산화질소 정량을 하였다. 결과: 2K1C 및 DOCA-salt 흰쥐에서 실험군은 각각의 대조군에 비해 유의하게 높은 혈압을 보였다. 두 고혈압군에서 모두 적출 대동맥 표본의 bNOS 및 ecNOS 단백 발현이 감소되었다. iNOS 단백은 DOCA-salt 고혈압에서 변화를 보이지 않으나 2K1C 고혈압에서는 역시 감소를 보였다. 혈관조직내 산화질소 함량은 두 고혈압에서 모두 유의하게 감소되었다. 결론: 2K1C 및 DOCA-salt 고혈압에서 혈관의 NOS 발현과 산화질소 함량이 감소되어 있으며 이는 고혈압의 유지 기전에 공헌하리라 추측되었다.

• 한방안이비인후피부과학회지
• /
• 제35권1호
• /
• pp.1-10
• /
• 2022

Objectives : This study was conducted to confirm the anti-inflammatory effects of Aster glehni Water extracts. Methods : In this study, MTT assay was performed to detect cell viability. To evaluate the anti-inflammatory effects of Aster glehni Water extracts, we examined NO production in LPS-induced macrophages. Expressions of iNOS, COX-2, ERK, p38, JNK were also investigated by using western blot assay. Results : Aster glehni Water extracts have no cytotoxicity at 15.625-1,000㎍/㎖ in RAW 264.7 cells. Aster glehni Extracts inhibited the NO production in a dose-dependent manner in RAW 264.7 cells treated with LPS. Pretreated 250, 500, 1,000㎍/㎖ of Aster glehni water extracts had significantly suppressed expression levels of iNOS, COX-2, p-ERK, p-p38, p-JNK. Conclusions : These results suggest that Aster glehni Water extracts have anti-inflammatory effects and can be used for various inflammatory skin diseases.

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

• Natural Product Sciences
• /
• 제20권4호
• /
• pp.227-231
• /
• 2014

Cimicifuga heracleifolia extract (CHE) was investigated for its effects on the release of nitric oxide (NO) and at the level of inducible nitric oxide synthase (iNOS) gene expression in mouse macrophages. We found that C. heracleifolia elicited a dose-dependent increase in NO production and the level of iNOS mRNA. Since, iNOS transcription has been shown to be under the control of the transcription factor $NF-{\kappa}B$, the effects of CHE on $NF-{\kappa}B$ activation were examined. Transient expression assays with $NF-{\kappa}B$ binding sites linked to the luciferase gene revealed that the increased level of iNOS mRNA, induced by CHE, was mediated by the $NF-{\kappa}B$ transcription factor complex. By using DNA fragments containing the $NF-{\kappa}B$ binding sequence, CHE was shown to activate the protein/DNA binding of $NF-{\kappa}B$ to its cognate site, as measured by electrophoretic mobility shift assay. These results demonstrate that C. heracleifolia stimulates NO production and is able to up-regulate iNOS expression through $NF-{\kappa}B$ transactivation.

• Journal of Acupuncture Research
• /
• 제21권3호
• /
• pp.107-119
• /
• 2004

Objective : The purpose of this study was to investigate the effect of Bee Venom and melittin on the lipopolysaccharide(LPS) and sodium nitroprusside(SNP)-induced expressions of Cell viability, nitric oxide(NO), inducible nitric oxide synthase(iNOS), extra-signal response kinase(ERK), jun N-terminal Kinase(JNK) and p38 kinase(p38)- mitogen activated protein kinase(MAPK) Family- in RAW 264.7 cells, a murine macrophage cell line. Methods : The expressions of cell viability by MTT assay, NO by Nitrite assay and iNOS, ERK, JNK and p38 were determined by Western blotting. Results : 1. Compared with the control group, 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin increased cell viability of RAW 264.7 induced by LPS and SNP significantly respectively. 2. Compared with the control group, 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin inhibited expression of NO induced by LPS and SNP significantly respectively. 3. Compared with the control group, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin inhibited expression of iNOS induced by LPS significantly and 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin inhibited expression of iNOS induced by SNP significantly. 4. Compared with the control group, the expression of ERK induced by LPS and SNP decreased significantly in the treatment groups of $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin, which of p-ERK by LPS also did in 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin, but which of p-ERK by SNP did not decrease. 5. Compared with the control group, the. expression of JNK induced by LPS and SNP decreased significantly in the treatment groups of 5, $10{\mu}g/m{\ell}$ melittin, which of p-JNK by LPS in 5, $10{\mu}g/m{\ell}$ melittin and by SNP in $1{\mu}g/m{\ell}$ bee venom and $10{\mu}g/m{\ell}$ melittin decreased significantly. 6. Compared with the control group, the expression of p38 induced by LPS did not have significant difference, which induced by SNP decreased significantly in the treatment groups of 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin. p-p38 induced by LPS decreased significantly in the treatment group of $10{\mu}g/m{\ell}$ of melittin, which induced by SNP also decreased significantly in 0.5, 1, $5{\mu}g/m{\ell}$ bee venom and 5, $10{\mu}g/m{\ell}$ melittin.

• Natural Product Sciences
• /
• 제18권3호
• /
• pp.147-152
• /
• 2012

Ergosta-4,6,8(14),22-tetraen-3-one (1), ergosta-7,24(28)-dien-3-ol (2), and 5,8-epidioxyergosta-6,22-dien-3-ol(3) were isolated from the fruit body of Phellinus pini. Their structures were based on spectroscopic methods including IR, MS, and NMR (1D and 2D). These compounds were screened for their ability to inhibit nitric oxide (NO) production in LPS-activated RAW 264.7 cells. Compounds 1, 2, and 3 reduced NO production in the assay with $IC_50$ values of 29.7 ${\mu}M$ (1), 15.1 ${\mu}M$ (2), and 18.4 ${\mu}M$ (3) respectively. They also suppressed the expression of protein and m-RNA of iNOS and COX-2 in a dose dependent manner by western blot analysis and RT-PCR experiment in LPS-activated microglial cells.

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.78.1-78.1
• /
• 2003

Sauchinone, a known lignan, was isolated from the root of Saururus chinensis as an active principle responsible for inhibiting the production of NO in LPS-stimulated RAW264.7 cells by activity-guided fractionation. Sauchinone dose-dependently inhibited not only the production of NO, but also the expression of iNOS mRNA and protein in LPS-stimulated RAW 264.7 cells. Furthermore, sauchinone prevented LPS-induced NF-kB activation, which is known to play a critical role in iNOS expression, assessed by a reporter assay under the control of NF-kB. (omitted)

• 대한약학회:학술대회논문집
• /
• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
• /
• pp.194.4-195
• /
• 2003

Two phenolic flavonoids were isolated from the traditional medicine of Eastern Asia, Caesalpinia sappan L (Leguminosae). Brazilin (1) showed a significant inhibitory activity against inducible Nitric Oxide Synthase (iNOS) in lipopolysaccharides (LPS)-induced macrophage RAW 264.7 cells with an IC$\sub$50/ value of 1.68 mg/ml, which is more potent than the positive control, L-N$\^$6/-(1-iminoethyl)lysine (IC$\sub$50/ 3.49 mM). On the other hand, caesalpine J (2) was found to be inactive in the present iNOS assay system despite of their structural similarities. (omitted)

• 대한본초학회지
• /
• 제21권2호
• /
• pp.135-142
• /
• 2006

Objectives : On-Bi-Tang(OB) has been prescribed Chinese traditional medicine for the treatment of inflammatory disease such as chronic renal failure. In this study, we investigated the anti-inflammatory effect of OB extract in the BV2 murine microglial cells. Methods : After the water extract of OB was treated in BV2 cells, murine microglial line, the cell viability was measured by MTT assay. The production of nitric oxide (NO) and $TNF-{\alpha}$ was determined based on Griess reagent and enzyme linked immunosorbant assay (ELISA). mRNA expression of inducible nitric oxide synthase (iNOS) and $TNF-{\alpha}$ was analyzed by RT-PCR. Results : OB extract significantly inhibited the LPS-induced production of NO and TNF-a in BV2 cells. OB extract also suppressed the mRNA expression of iNOS and $TNF-{\alpha}$ in BV2 cells activated with LPS. Conclusion : These data suggests that OB extract may have the anti-inflammatory effect through the modulation of NO production and inflammatory cytokine such as $TNF-{\alpha}$.

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