• IMMUNE NETWORK
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• 제5권1호
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• pp.45-49
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• 2005

Background: Inflammation in the brain has known to be associated with the development of a various neurological diseases. The hallmark of neuro-inflammation is the activation of microglia, brain macrophage. Pro-inflammatory compounds including nitric oxide (NO) are the main cause of neuro-degenerative disease such as Alzheimer's disease (AD) which is resulted in cell death. Among those pro-inflammatory compounds, NO contributes to the cell death by directly or indirectly. Methods: In the study, we examined whether ursodeoxycholic acid (UDCA), a non-toxic hydrophilic bile acid, inhibits the NO production by a direct method using Griess reagent and by RT-PCR in the gene expression of inducible nitric oxide synthase (iNOS). In signal transduction, we also examined the NF-${\kappa}B$ (p65/p50), IKK, and I ${\kappa}B$, which are associated with the expression of iNOS gene using western blots. Results: In the present study, we found that UDCA effectively inhibited NO production in BV-2 microglial cell, and NF-${\kappa}B$ activation was reduced by suppressing IKK gene expression and by increasing the I${\kappa}B$ in cytosol comparing those to the positive control LPS. Conclusion: Taken together, these data suggested that UDCA may playa crucial role in inhibiting the NO production and the results imply that UDCA suppresses a cue signal of the microglial activation via stimulators, such as ${\beta}$-amyloid peptides which are known to stimulate microglia in AD pathogenesis.

• 생명과학회지
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• 제27권1호
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• pp.8-14
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• 2017

치자 열매는 아시아에서 음식과 옷의 염료로 사용되어 왔다. 본 연구에서는 BV-2 소교세포에서 치자열매 열수추출물(GJ)의 항염증 효과를 관찰하고 그 작용 기전을 연구하였다. GJ는 세포에 독성을 유도하지 않으면서 lipopolysaccharide로 인한 nitric oxide (NO) 분비와 inducible nitric oxide synthase (iNOS) 생성 및 활성산소 생성을 억제하였다. 또한 GJ는 농도의존적으로 heme oxygenase-1 (HO-1)의 발현을 유도하였다. 더군다나 HO-1 siRNA를 처리했을 때는 GJ가 iNOS의 발현을 억제하지 못하였다. GJ는 HO-1의 발현에 관여하는 전사인자인 nuclear factor E2-related factor 2를 핵으로 이동시켰다. 또한 GJ에 의한 HO-1의 발현은 phosphatidylinositol 3-kinase(PI-3K) 및 p38 kinase 억제제에 의해 감소되었으며, GJ가 Akt와 p38 kinase의 인산화를 유도하였다. 이상의 결과를 종합해보면, GJ는 PI3K/Akt 및 p38 신호전달과정을 통해 HO-1의 발현을 유도함으로써 NO와 같은 염증매개물질의 생성을 억제한다는 것을 알 수 있다. 이러한 연구결과는 치자열매가 신경염증을 억제하는 새로운 기전을 밝힌 것이다.

• 대한약학회:학술대회논문집
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• 대한약학회 2003년도 Proceedings of the Convention of the Pharmaceutical Society of Korea Vol.2-2
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• pp.205.1-205.1
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• 2003

Several lines of recent evidences have shown that several pro-inflammatory genes or mediators, such as inducible nitric oxide synthase (iNOS)are strongly expressed in the ischemic brain. Inflammation is now recognized as a significant contributing mechanism in cerebral ischemia because anti-inflammatory compounds or inhibitors of iNOS have been proven to reduce ischemic brain damage. In iNOS assay, hexane fraction of M61 inhibited NO (iNOS IC50, 0.7${\mu}$g/ml). In vivo study was carried out to evaluate neuroprotective effect of hexane fraction of M61 after transient global ischemia using Mongolian gerbil ischemia model. (omitted)

• Archives of Pharmacal Research
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• 제27권1호
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• pp.111-117
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• 2004

Hepatic microcirculatory failure is a major component of reperfusion injury in the liver. Recent data provided some evidence that endothelium-derived vasoconstrictors and vasodilators may be functionally important to the control of the total hepatic blood flow under these conditions of circulatory failure. Since Kupffer cells provide signals that regulate the hepatic response in ischemia/reperfusion (I/R), the aim of this study was to investigate the role of Kupffer cells in the I/R-induced imbalance of vasoregulatory gene expression. Rats were subjected to 60 min hepatic ischemia, followed by 5 h of reperfusion. The Kupffer cells were inactivated by gadolinium chloride ($GdCl_3$, 7.5 mg/kg body weight, intravenously) 1 day prior to ischemia. Liver samples were obtained 5 hrs after reperfusion for RT-PCR analysis of the mRNA for genes of interest: endothelin-1 (ET-1), its receptors $ET_A and ET_B$, endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS) and heme oxygenase-1 (HO-1). ET-1 mRNA expression was increased by I/R. mRNA levels for $ET_A$ receptors showed no change, whereas $ET_B$ receptor transcripts increased in the I/R group. The increases in ET-1 and $ET_B$ mRNA were not prevented by the $GdCI_3$ pretreatment. The mRNA levels for iNOS and eNOS significantly increased within the I/R group with no significant difference between the I/R group and the $GdCl_3$-treated I/R group. HO-1 mRNA expression significantly increased in the I/R group and this increase was attenuated by $GdCI_3$. In conclusion, we have demonstrated that an imbalance in hepatic vasoregulatory gene expression occurs during I/R. Our findings suggest that the activation of Kupffer cells is not required for I/R-induced hepatic microvascular dysfunction.

• Archives of Pharmacal Research
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• 제27권2호
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• pp.225-231
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• 2004

This study was designed to investigate the effect of Trolox, a hydrophilic analogue of vitamin E, on the alteration of vasoregulatory gene expression during hepatic ischemia and reperfusion (I/R). Rats were subjected to 60 min of hepatic ischemia in vivo. The rats were treated intravenously with Trolox (2.5 mg/kg) or the vehicle as a control 5 min before reperfusion. Liver samples were obtained 5 h after reperfusion for a RT-PCR analysis on the mRNA for the genes of interest. These mRNA peptides are endothelin-1 (ET -1), potent vasoconstrictor peptide, its receptor $ET_A$ and $ET_B$, vasodilator endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), tumor necrosis factor-$\alpha$ (TNF-$\alpha$) and cyclooxygenase-2 (COX-2). It was seen that serum alanine aminotransferase and lipid peroxi-dation levels were markedly increased after I/R and Trolox significantly suppressed this increase. In contrast, the glutathione concentration decreased in the I/R group, and this decrease was inhibited by Trolox. ET-1 mRNA expression was increased by I/R, an increase which was prevented by Trolox. The mRNA levels for $ET_A$ receptor was significantly decreased, whereas ET$_{B}$ receptor transcript increased in the I/R group. The increase in $ET_A$ was prevented by Trolox. The mRNA levels for iNOS and HO-1 significantly increased in the I/R group and Trolox attenuated this increase. There were no significant differences in eNOS mRNA expression among any of the experimental groups. The mRNA levels for COX-2 and TNF-$\alpha$ significantly increased in I/R group and Trolox also attenuated this increase. Our findings suggest that I/R induces an imbalanced hepatic vasoregulatory gene expression and Trolox ameliorates this change through its free radical scavenging activity.y.

• Journal of Ginseng Research
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• 제37권1호
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• pp.54-63
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• 2013

Ginsenoside Rd is a primary constituent of the ginseng rhizome and has been shown to participate in the regulation of diabetes and in tumor formation. Reports also show that ginsenoside Rd exerts anti-oxidative effects by activating anti-oxidant enzymes. Treatment with ginsenoside Rd decreased nitric oxide and prostaglandin $E_2$ ($PGE_2$) in lipopolysaccharides (LPS)-challenged RAW264.7 cells and in ICR mouse livers (5 mg/kg LPS; LPS + ginsenoside Rd [2, 10, and 50 mg/kg]). Furthermore, these decreases were associated with the down-regulations of inducible nitric oxide synthase (iNOS) and cyclooxygenase (COX)-2 and of nuclear factor (NF)-${\kappa}B$ activity in vitro and in vivo. Our results indicate that ginsenoside Rd treatment decreases; 1) nitric oxide production (40% inhibition); 2) $PGE_2$ synthesis (69% to 93% inhibition); 3) NF-${\kappa}B$ activity; and 4) the NF-${\kappa}B$-regulated expressions of iNOS and COX-2. Taken together, our results suggest that the anti-inflammatory effects of ginsenoside Rd are due to the down-regulation of NF-${\kappa}B$ and the consequent expressional suppressions of iNOS and COX-2.

• 한국미생물·생명공학회지
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• 제45권1호
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• pp.35-42
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• 2017

본 연구에서, 비발효(ARE) 및 발효(ARE-F) 배초향 잎 열수 추출물의 피부 항산화 및 항염증 효능을 비교함으로써 프로바이오틱 발효의 효과를 검토하였다. ARE-F는 Lactobacillus rhamnosus HK-9 발효에 의하여 ARE로부터 제조되었다. In vitro 항염증 효능은 지질다당류(LPS)에 의하여 자극된 HaCaT 각질세포에서 일산화질소(NO), 활성산소종(ROS) 및 유도형 일산화질소합성효소(iNOS) 분석에 의하여 평가되었다. In vitro antiradical 활성은 2,2-diphenyl-2-picrylhydrazyl radical (DPPH) 소거 측정법에 의하여 평가되었다. LPS 자극에 의하여 증가된 NO, ROS 및 iNOS 수준에 미치는 ARE-F의 감소효과가 ARE에 의한 효과보다 현저히 강하였다. 그러나, ARE와 ARE-F의 DPPH 소거 효능에는 차이가 발견되지 않았다. ARE-F가 증강된 피부 항산화 및 항염증 효능을 갖고 있는데, 이 결과는 프로바이틱세균 발효가 배초향 잎의 일부 약리학적 효능을 증가시키는 효과적인 방법일 수 있음을 암시해 주고 있다. 요약하면, 배초향 잎 추출물의 피부항산화 및 항염증 활성이 프로바이오틱 세균인 L. rhamnosus HK-9에 의한 발효에 의하여 증강한다.

• IMMUNE NETWORK
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• 제4권2호
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• pp.108-115
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• 2004

Background: Production of nitric oxide (NO) by inducible NO synthase (iNOS) has been implicated in the pathology of autoimmune disease. It is unknown whether iNOS expression is increased within testes and whether iNOS and NO have essential roles in the pathogenesis of EAO. Methods: EAO was induced in guinea pig testes at 17 days after secondary immunization by administration of crude extract (CE) and purified glycoprotein 1 (GP1) from normal guinea pig testes. iNOS gene expression was assessed by RT-PCR and Northern blot analysis in testes. Localization of iNOS and Mac-1 and the indicator of NO-mediated tissue injury, nitrotyrosine, were detected in the testicular lesion by immunohistochemistry. Results: In control testes, inflammation and iNOS gene expression were not detected, whereas, in CE- and GP1-injected testes, inflammation and marked iNOS gene expression were evident at day 17 after secondary immunization. Immunohistochemistry of Mac-1 showed the colocalization with iNOS protein and nitrotyrosyl proteins in intertubules, suggesting that NO produced by infiltrated macrophages may be involved in inflammatory lesions of intertubules. Intraperitoneal administration of aminoguanidine significantly prevented EAO with reduction of inflammation, iNOS expression and nitrotyrosine formation. Conclusion: These results suggest that NO production by macrophages may be important in the pathogenesis of CE- and GP1-induced EAO. Furthermore, this study demonstrated the therapeutic potential of iNOS inhibitor in the treatment of inflammatory and autoimmune mediated-diseases.

• The Korean Journal of Physiology and Pharmacology
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• 제5권6호
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• pp.521-527
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• 2001

Diclofenac, a phenylacetic acid derivative, is a widely used non-steroidal anti-inflammatory drug (NSAID) to provide effective relief of inflammation and pain. Nitric oxide (NO) synthesized by inducible nitric oxide synthase (iNOS) has been implicated as a mediator of inflammation. We examined the inhibitory effects of diclofenac on the induction of iNOS in RAW 264.7 macrophages which were activated with lipopolysaccharide (LPS) plus interferon-gamma $(IFN-{\gamma}).$ Treatment of RAW 264.7 cells with diclofenac and other NSAIDs (aspirin and indomethacin) significantly inhibited NO production and iNOS protein expression induced by LPS plus $IFN-{\gamma}.$ Also, diclofenac but not aspirin and indomethacin, inhibited iNOS mRNA expression and nuclear factor-kappa B $(NF-{\kappa}B)$ binding activity concentration-dependently. Furthermore, transfection of RAW 264.7 cells with iNOS promoter linked to a CAT reporter gene revealed that only diclofenac inhibited the iNOS promoter activity induced by LPS plus $IFN-{\gamma}$ through the $NF-{\kappa}B$ sites of iNOS promoter. Taken together, these suggest that diclofenac may exert its anti-inflammatory effect by inhibiting iNOS gene expression at the transcriptional level through suppression of $NF-{\kappa}B$ activation.

• 한국방사선학회논문지
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• 제13권1호
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• pp.133-140
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• 2019

중성지방(Triglyceride, TG)는 죽상동맥경화증과 같은 혈관의 만성 염증성 병변을 유발하는 인자 중 하나이다. 종양괴사인자-알파 ($TNF-{\alpha}$), 인터루킨-1 베터 ($IL-1{\beta}$)와 같은 염증성 사이토카인은 염증 질환의 주요 요인으로 염증 부위에 T 림프구, 단핵구등의 면역 세포의 침윤을 유도하거나 세포 및 조직 괴사를 일으킴으로써 질병을 더욱 악화시킨다. 본 연구에서는 혈관 염증에 관여하는 Jurkat T 림프구와 U937 단핵구에 TG를 처리하였을 때 $TNF-{\alpha}$와 $IL-1{\beta}$의 발현에 미치는 영향을 조사하고자 했다. Jurkat T 세포에서 TG에 의해 $TNF-{\alpha}$의 mRNA 발현이 증가하였고, U937 단핵구에서는 TG에 의해 $TNF-{\alpha}$와 $IL-1{\beta}$ 모두 mRNA 발현이 증가하였다. 또한 유도성 산화질소합성효소(inducible nitric oxide synthase, iNOS)가 TG에 의한 $TNF-{\alpha}$와 $IL-1{\beta}$의 발현 증가에 관여하는지 확인하기 위해 iNOS 억제제인 W1400을 세포에 전처리하여 iNOS의 활성을 차단하였다. 그 결과, W1400을 전처리한 세포에서는 TG에 의한 $TNF-{\alpha}$ 및 $IL-1{\beta}$ mRNA 양이 대조군과 유사하게 낮은 수준으로 관찰되었다. 이는 혈관 내 TG의 증가가 T 림프구와 단핵구를 자극하여 iNOS 신호를 거쳐 염증성 사이토카인을 분비시키고 혈관염증질환을 발생하는데 관여하는 것을 확인시켜주었다. 결론적으로, 중성지방이 염증성 병변을 악화시키는데 있어 iNOS의 활성이 사이토카인 분비 등에 작용하며 병변을 더욱 악화시키는데 기여할 수 있다. 반면, iNOS 발현을 조절하여 고지혈증 환자의 치료에 유효한 표적 물질로 이용될 가능성이 있다고 사료된다.