• 대한약학회:학술대회논문집
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• 대한약학회 2002년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2
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• pp.321.2-321.2
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• 2002

Possible role of anti-inflammatory effects of a new iridoids, patridoid I. II and II-A which were isolated from Patrinia saniculaefolia. examined by assessing their effects on tumor necrosis factor $\alpha$ (TN F$\alpha$) and 2 enzymes, inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in the lipopolysaccaride (LPS)-stimulated murine macrophage-like cell line RAW 264.7. Among them. patridoid II consistently inhibited the production of TNF$\alpha$ and NO production in a dose dependent manner. But patridoid I and patrioid ll isomer palrioid ll-A. these compounds very weakly inhibited NO producion. Moreover. treatment of macrophage with these compounds, the decrease in NO products was accompanied by a decrease in iNOS protein level as assessed by Western Blot. But these compounds did not affect COX-2 protein expression in LPS-stimulated macrophage. Our results suggest that patridoid ll could become a leading compound for developing a novel of anti-inflammalory drugs.

• 한국식품영양과학회지
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• 제29권2호
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• pp.342-348
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• 2000

In this experiment, we show the effects of herbal plant extracts on the production of nitric oxide (NO) and TNF-$\alpha$. The extracts of Angelica gigas, Astragalus membranaceus, Acanthopanax sessiliflorus and Houttuynia cordata had no effect on NO synthesis by itself in mouse macrophage cell line (RAW264.7). However, the stimulation with these extracts in the presence of murine interferon-${\gamma}$(mIFN-${\gamma}$) resulted in increased NO synthesis. When these extracts were used in combination with mIFN-${\gamma}$, there were a marked cooperative induction of NO and TNF-$\alpha$ synthesis in a dose-dependent manner. The same results were obtained in the mouse peritoneal macrophages used. The optimal concentration of these extracts on NO synthesis was shown at 100$\mu\textrm{g}$/mL with 100U/mL of mIFN-${\gamma}$. NO synthesis was inhibited by NG-monomethyl-L-arginine. When cell lines were treated with extracts, the expression of inducible NO synthetase (iNOS) was markedly increased in RT-PCR analysis. In addition, synergy between mIFN-${\gamma}$ and extracts was dependent on extracts-induced tumor necrosis factor-$\alpha$(TNF-$\alpha$). These results suggest that water extracts of herbal plants can induce iNOS, NO and TNF-$\alpha$ synthesis of mouse macrophage cell line (RAW264.7) and peritoneal macrophages in combination with mIFN-${\gamma}$.

• Journal of Acupuncture Research
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• 제25권3호
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• pp.95-106
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• 2008

Objectives : This study is to evaluate Effect of Hong Hua Za acupuncture anti-inflammatory on rheumatoid arthritis that caused by Lipopolysaccharide in mouse. Methods : Hong Hua Za(Carthami Tinctorii Fructus) acupuncture(HA) was prepared and tested for therapeutic potential of rheumatoid arthritis, by measuring the inhibition of inducible nitric oxide synthase (iNOS) expression, nitric oxide(NO) production in vitro experiment, by measuring Tumpr Necrosis Factor-$\alpha$(TNF-$\alpha$) & Nuclear Factor kappa B(NF-$\kappa$B) p65 activation, synovial hyperplasia, angiogenesis and fibrosis in synovial membrane of knee joint of mice. Results : HA showed therapeutic effects on Lipopolysaccharide(LPS)-induced rheumatoid arthritis. HA inhibited iNOS mRNA and NO in RAW264.7cell of LPS-induced rheumatoid arthritis in a dose dependent manner. HA also showed significant inhibition of TNF-$\alpha$ & NF-$\kappa$B p65, activation, synovial hyperplasia, angiogenesis and fibrosis in synovial membrane of knee joint of mice. Conclusions : These results suggest that HA has an therapeutic effects on LPS induced-rheumatoid arthritis by inhibiting TNF-$\alpha$ activation.

• Biomolecules & Therapeutics
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• 제23권2호
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• pp.110-118
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• 2015

According to the expansion of lifespan, neuronal disorder based on inflammation has been social problem. Therefore, we isolated shikonin from Lithospermum erythrorhizon and evaluated anti-inflammatory effects of shikonin in lipopolysaccharide (LSP)-stimulated BV2 microglial cells. Shikonin dose-dependently inhibits the expression of the proinflammatory mediators, nitric oxide (NO), prostaglandin $E_2$ ($PGE_2$), and tumor necrosis factor-${\kappa}B$ (TNF-${\alpha}$) as well as their main regulatory genes and products such as inducible NO synthase (iNOS), cyclooxygenase-2 (COX-2), and TNF-${\alpha}$ in LPS-stimulated BV2 microglial cells. Additionally, shikonin suppressed the LPS-induced DNA-binding activity of nuclear factor-${\kappa}B$ (NF-${\kappa}B$) to regulate the key regulatory genes of the proinflammatory mediators, such as iNOS, COX-2, and TNF-${\alpha}$, accompanied with downregulation of reactive oxygen species (ROS) generation. The results indicate that shikonin may downregulate the expression of proinflammatory genes involved in the synthesis of NO, $PGE_2$, and TNF-${\alpha}$ in LPS-treated BV2 microglial cells by suppressing ROS and NF-${\kappa}B$. Taken together, our results revealed that shikonin exerts downregulation of proinflammatory mediators by interference the ROS and NF-${\kappa}B$ signaling pathway.

• Clinical and Experimental Pediatrics
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• 제56권3호
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• pp.116-124
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• 2013

Purpose: Tumor necrosis factor (TNF)-${\alpha}$ is thought to contribute to pulmonary hypertension. We aimed to investigate the effect of infliximab (TNF-${\alpha}$ antagonist) treatment on pathologic findings and gene expression in a monocrotaline-induced pulmonary hypertension rat model. Methods: Six-week-old male Sprague-Dawley rats were allocated to 3 groups: control (C), single subcutaneous injection of normal saline (0.1 mL/kg); monocrotaline (M), single subcutaneous injection of monocrotaline (60 mg/kg); and monocrotaline + infliximab (M+I), single subcutaneous injection of monocrotaline plus single subcutaneous injection of infliximab (5 mg/kg). The rats were sacrificed after 1, 5, 7, 14, or 28 days. We examined changes in pathology and gene expression levels of TNF-${\alpha}$, endothelin-1 (ET-1), endothelin receptor A (ERA), endothelial nitric oxide synthase (eNOS), matrix metalloproteinase (MMP) 2, and tissue inhibitor of matrix metalloproteinase (TIMP). Results: The increase in medial wall thickness of the pulmonary arteriole in the M+I group was significantly lower than that in the M group on day 7 after infliximab treatment (P<0.05). The number of intraacinar muscular arteries in the M+I group was lower than that in the M group on days 14 and 28 (P<0.05). Expression levels of TNF-${\alpha}$, ET-1, ERA, and MMP2 were significantly lower in the M+I group than in the M group on day 5, whereas eNOS and TIMP expressions were late in the M group (day 28). Conclusion: Infliximab administration induced early changes in pathological findings and expression levels of TNF-${\alpha}$, and MMP2 in a monocrotaline-induced pulmonary hypertension rat model.

• 대한한방부인과학회지
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• 제20권3호
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• pp.111-128
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• 2007

목적 : 이 연구는 천식, 기관지염, 폐렴, 결핵, 산후감모 등의 호흡기 질환에 사용되는 가미지황탕(加味地黃場)의 항염작용(抗炎作用)의 효과에 대해 알아보기 위해 시행되었다. 방법 : 가미지황탕(加味地黃場)의 항염작용(抗炎作用)의 효과를 평가하기 위해 세포독성에 미치는 영향, NO, $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 생성량에 미치는 영향, $TNF-{\alpha}$, $IL-1{\beta}$, IL-6 유전자 발현에 미치는 영향, iNOS, COX-2 유전자 및 단백질 발현에 미치는 영향, $PGE_2$ 합성에 미치는 영향 및 COX-2, $NF-{\kappa}B$ 활성에 미치는 영향에 대한 실험평가를 하였다. 결과 : 가미지황탕(加味地黃場)은 MTT 분석을 통한 RAW 264.7 세포주의 생존력 평가에서 세포독성이 없었고, LPS로 유도된 RAW 264.7 세포주에서 NO, $TNF-{\alpha}$, $IL-1{\beta}$ 및 IL-6 생성량을 농도 의존적으로 억제하였다. 가미지황탕(加味地黃場)은 400 g/ml 농도에서 LPS로 유도된 RAW 264.7 세포주에 대해 $TNF-{\alpha}$, $IL-1{\beta}$ 및 IL-6 유전자 발현을 농도 의존적으로 억제하였고, LPS로 유도된 RAW 264.7 세포주에서 iNOS와 COX-2 유전자 및 단백질 발현은 농도 의존적으로 억제하였다. 또한 그 농도에 따라 $PGE_2$ 생성량이 현저하게 억제하였고, LPS로 유도된 COX-2 및 $NF-{\kappa}B$ 전사활성을 농도 의존적으로 억제함으로써 iNOS와 COX-2 유전자 발현을 억제하였다. 결론 : 이상의 실험을 통해 가미지황탕(加味地黃場)은 iNOS나 COX-2와 같은 Cytokine이 있는 효소에 의해 합성되고 천식에서 증가하는, 혈관과 기관지 긴장도와 관련 있는 NO와 $PGE_2$ 생성량을 억제하고, 염증과 관련된 $TNF-{\alpha}$, $IL-1{\beta}$, IL-6의 생성량을 억제하였다. 또한 $NF-{\kappa}B$ 활성을 억제함으로써 iNOS 및 COx-2 유전자 발현을 억제하였으므로 부인과 영역에 있어서도 산후감모, 만성해수 및 천식 등의 기관지의 염증질환에 응용할 수 있을 것으로 사료된다.

• Natural Product Sciences
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• 제16권4호
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• pp.265-270
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• 2010

The methanol extract of Zanthoxylum rhetsa (MZRR) were evaluated for its ability to suppress the formation of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) in lipopolysaccharide (LPS)-activated RAW 264.7 macrophages. MZRR presented an inhibition of LPS-induced production of nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) in RAW 264.7 macrophages. Western blotting and RT-PCR analyses demonstrated that MZRR significantly inhibited the protein and mRNA expressions of iNOS and COX-2 in LPS-activated macrophages in a dose-dependent manner. LPS-induced COX-2, iNOS, and nuclear factor kappa beta (NF-${\kappa}B$) activity were also decreased in the presence of MZRR. The production of tumor necrosis factor-$\alpha$ (TNF-$\alpha$), the mRNA expression levels of pro-inflammatory cytokines, including TNF-$\alpha$ and IL-$1{\beta}$, were reduced after MZRR administration in a dose dependent-manner. These results suggest that the MZRR extract involved in the inhibition of iNOS and COX-2 via the NF-${\kappa}B$ pathway, revealing a partial molecular basis for anti-inflammatory properties of the MZRR extract.

• Journal of Acupuncture Research
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• 제25권5호
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• pp.105-116
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• 2008

Objectives : The objective of this study is to investigate the suppressing effect of the cervi pantotrichum cornu pharmacopuncture on the expression of iNOS mRNA and production of NO in synoviocytes from artificially arthritis-induced mice. Methods : In vitro test, synoviocytes extracted from a knee joint of a mouse were cultivated, and the herbal extract of cervi pantotrichum cornu($0.4mg/m{\ell}$, $0.6mg/m{\ell}$, $0.8mg/m{\ell}$, and $1.0mg/m{\ell}$) was added into the wells of synoviocytes to suppress the expression of iNOS mRNA and production of NO. In vivo test, each ten mice were allocated into three groups; Normal group, CIA-elicitated group(CIA), and group treated with cervi pantotrichum cornu pharmacopuncture after CIA elicitation(CCA). The extract of cervi pantotrichum cornu was injected into the acupoint of $SP_{10}$ to observe the changes of foot thickness in mice and the suppression of MIF, TNF-$\alpha$, NF-${\kappa}B$ p65, and iNOS. Results : In vitro test, the expression of iNOS mRNA and production of NO were dose-dependently decreased in the wells of synoviocytes treated with PMA. In vivo test, the suppression of MIF, TNF-$\alpha$, NF-${\kappa}B$ p65, and iNOS was clearly shown in the pieces of the synovial joint treated with the extract of cervi pantotrichum cornu. The foot thickness also decreased dose-dependently. Conclusions : It is speculated that the cervi pantotrichum cornu pharmacopuncture can be applicable to the therapy of rheumatoid arthritis by suppressing the expression of iNOS mRNA and production of NO.

• 디지털융복합연구
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• 제15권11호
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• pp.513-522
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• 2017

Murine macrophage cell line인 RAW 264.7 대식세포에 참당귀(AG), 중국당귀(AS), 일당귀(AA)를 6시간과 24시간을 반응시킨 뒤 시간별로 나타나는 inflammation 반응을 확인하고자 하였다. LPS-induced RAW 264.7 대식세포에 AG, AS, AA를 6시간 반응 시킨 경우 $IL-1{\beta}$, $TNF-{\alpha}$, iNOS의 mRNA 발현량을 증가시켰고, AA에서 더 높은 Anti-inflammatory effect를 확인하였다. AG, AS, AA가 RAW 264.7 대식세포의 cell viability에 미치는 영향을 확인하기 위해 MTS Assay(24시간)를 수행한 결과 $1,600{\mu}g/ml$ 농도에서 모두 증가시켰다. AG, AS, AA가 LPS를 처리하지 않은 RAW 264.7 대식세포에 6시간동안 면역반응에 미치는 영향을 확인한 결과 AG와 AA를 $200{\mu}g/ml$ 농도로 처리한 RAW 264.7 대식세포에서 $IL-1{\beta}$, $TNF-{\alpha}$, iNOS의 발현이 증가되었다. 이 연구를 통해 AG, AS, AA는 RAW 264.7 대식세포에 LPS를 처리하였을 때 inflammation 반응을 촉진하며, 24시간 뒤 inflammation의 억제를 유도한 결과를 얻을 수 있었다. 향후 참당귀, 중국당귀, 일당귀 열수 추출물이 대식세포의 다양한 염증 반응 메커니즘에 미치는 작용을 확인하는 연구가 필요하다.

• Molecular & Cellular Toxicology
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• 제1권3호
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• pp.196-202
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• 2005

Nitric oxide (NO) is a small, diffusible, and highly reactive molecule, which plays dichotomous regulatory roles under physiological and pathological conditions. NO promotes apoptosis in some cells, and inhibits apoptosis in other cells. In the present study, we attempted to characterize the NO signaling pathway and cellular response in PC12 cells treated with cytokines. $IFN{\gamma}\;and\;TNF{\alpha}$ treatment resulted in a synergistic increase of nitrite accumulation, with the induction of inducible nitric oxide synthase (iNOS) in the PC12 cells. Moreover, as nitrite concentration increased, cell viability decreased. In order to explore MAP kinase involvement in nitric oxide production resultant from $IFN{\gamma}\;and\;TNF{\alpha}$ stimulation, we measured the activation of MAP kinase using specific MAP kinase inhibitors. PC12 cells pretreated with SB203580, a p38 MAP kinase-specific inhibitor, resulted in the inhibition of iNOS expression and NO production. However, PD98059, an ERK/MAP kinase-specific inhibitor, was not observed to exert such an effect. In addition, Stat1 activated by $IFN{\gamma}\;and\;TNF{\alpha}$ was interacted with p38 MAPK. These data suggest that p38 MAP kinase mediates cytokine-mediated iNOS expression in the PC12 cells, and Jak/Stat pathway interferes with p38 MAPK signaling pathway.