• The Korean Journal of Physiology and Pharmacology
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• 제17권4호
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• pp.315-320
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• 2013

Here, we show that radicicol, a fungal antibiotic, resulted in marked inhibition of inducible nitric oxide synthase (iNOS) transcription by the pancreatic beta cell line MIN6N8a in response to cytokine mixture (CM: TNF-${\alpha}$, IFN-${\gamma}$, and IL-$1{\beta}$). Treatment of MIN6N8a cells with radicicol inhibited CM-stimulated activation of NF-${\kappa}B$/Rel, which plays a critical role in iNOS transcription, in a dose-related manner. Nitrite production in the presence of PD98059, a specific inhibitor of the extracellular signal-regulated protein kinase-1 and 2 (ERK1/2) pathway, was dramatically diminished, suggesting that the ERK1/2 pathway is involved in CM-induced iNOS expression. In contrast, SB203580, a specific inhibitor of p38, had no effect on nitrite generation. Collectively, this series of experiments indicates that radicicol inhibits iNOS gene expression by blocking ERK1/2 signaling. Due to the critical role that NO release plays in mediating destruction of pancreatic beta cells, the inhibitory effects of radicicol on iNOS expression suggest that radicicol may represent a useful anti-diabetic activity.

• 한국독성학회:학술대회논문집
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• 한국독성학회 2002년도 Current Trends in Toxicological Sciences
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• pp.88-88
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• 2002

Cyclooxygenase(COX) metabolizes arachidonic acid to prostaglandins and thromboxanes. It has been reported that there is 'cross-talk' between COX-2 and nitric oxide synthase(NOS). Stimulation of eNOS of estrogen fed animal heart was not accompanied by the increase in COX-2 expression.(omitted)

• 한국미생물·생명공학회지
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• 제44권4호
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• pp.571-576
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• 2016

본 연구에서는 섬바디 나물의 항염증 효과를 알아보기 위하여 LPS로 염증을 유도한 Raw 264.7 세포에 대한 섬바디나물 80% 에탄올 추출물의 효과를 살펴보았다. 섬바디 나물 추출물을 LPS로 유도된 Raw 264.7 대식세포에서 전염증성인자(iNOS, COX-2)들을 생성하여 측정하였다. 섬바디 나물 추출물의 대식세포에서의 세포 독성 측정을 MTT를 수행하였다. 섬바디 나물 추출물의 세포 독성을 측정한 결과, $1,000{\mu}g/ml$의 농도에서 100% 이상의 세포 생존율을 보였다. 섬바디 나물 추출물의 50, 100, $500{\mu}g/ml$ 농도에서 iNOS와 COX-2의 단백질 발현 억제 효과를 측정하기 위해 western blot을 통해 측정하였고, 양성대조군으로는 ${\beta}-actin$을 사용하였다. 그 결과, 섬바디 나물 추출물을 western blot을 통해 측정한 iNOS, COX-2의 단백질 발현 억제 효과는 $500{\mu}g/ml$ 농도에서 각각 56%, 61.6%로 감소하였다. 섬바디 나물 추출물의 50, 100, $500{\mu}g/ml$ 농도에서 iNOS, COX-2의 mRNA의 발현억제 효과를 측정하기 위해 RT-PCR을 통해 측정하였으며, 양성 대조군으로 GAPDH를 사용하였다. 그 결과, 섬바디 나물 추출물을 RT-PCR을 통해 iNOS, COX-2의 mRNA 발현 억제 효과를 측정한 결과는 $500{\mu}g/ml$에서 각각 77.9%, 83.3%로 감소하였다. 이를 통해, 섬바디 나물 추출물은 염증을 억제시켜 주는 가능성이 있는 항염증 물질로써의 효과가 있을 것으로 보여진다.

• 대한한방내과학회지
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• 제39권1호
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• pp.1-8
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• 2018

Objectives: This study seeks to investigate the effects of Gamidohongsamul-tang (GDT) on the Gene expression levels of eNOS, KLF2, ICAM-1 and VCAM-1 in HUVEC cells. Methods: HUVEC cells were treated at a concentration of 1, 10, 100 (${\mu}g/ml$) of Gamidohongsamul-tang (GDT). To measure the NOS, KLF2, ICAM-1 and VCAM-1 gene expression in HUVEC cells, the synthesized cDNA was subjected to polymerase chain reaction (PCR) and electrophoresis was performed to verify gene expression level. Results: 1. GDT significantly increased eNOS and KLF2 gene expression. 2. GDT significantly reduced ICAM-1 and VCAM-1 gene expression. Conclusions: These experiments suggest that Gamidohongsamul-tang (GDT) regulates gene expression related with anti-dyslipidemic effects in HUVEC cells. In order to clinically apply this to diseases related to dyslipidemia, such as cardiovascular disease, additional in vivo experiments are needed to verify the anti-dyslipidemic effects of GDT.

• Archives of Pharmacal Research
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• 제25권2호
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• pp.151-153
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• 2002

Three pseudoguaianolide type sesquiterpenes, bigelovin(1), 2,3-dihydroaromaticin (2), and ergolide (3) were isolated as inhibitory constituents against inducible nitric oxide synthase (iNOS) from the flowers of Inula britannica var. chinensis. Bigelovin (1) exhibited a highly potent inhibitory activity on lipopolysaccharide (LPS)-induced iNOS in murine macrophage RAW 264.7 cells with an $IC_{50}$ value of 0.46 mM, which is about 8 times more potent than the known selective inhibitor of iNOS, $L-N^6-(1-iminoethyl)Iysine{\;}(IC_{50}{\;}3.49{\;}{\mu}M)$. 2,3-Dihydroaromaticin (2) and ergolide (3) also exhibited potent inhibitory activities on LPS-induced iNOS with $IC_{50}$ values of 1.05 and $0.69{\;}{\mu}M$, respectively.

• 한국산림과학회지
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• 제84권1호
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• pp.77-86
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• 1995

임목을 대상으로 삽입된 외래 유전자의 공간적, 시기별 발현 특성을 이해하기 위한 기초연구로서 온실에서 생육 중인 형질전환된 2년생 잡종 포플러 (Populus alba X P. grandidentata) Hansen 클론을 대상으로 삽입된 외래 유전자의 발현정도를 각 기관별로 조사하였다. Agrobacterium binary vector pRT45, pRT102 및 pRT104에 의해서 형질전환된 3계통의 형질전환체 Tr15, Tr345, Tr665 모두는 선발가능한 표식 유전자로서 nos promoter-NPT II 유전자가 대상 식물체의 genome에 삽입되어 있으며, 그외에, pin2 promoter-CAT 유전자(pRT45), nos promoter-PIN2 유전자(pRT102), Cauliflower Mosaic Virus 35s promoter-PIN2 유전자(pRT104)가 3계통의 형질전환체에 제각각 삽입되어 있는 잡종 포플러이다. 이들 3계통의 형질전환 포플러 식물체의 DNA를 PCR 검정 기법을 이용하여 분석해 본 결과 선발 가능한 표식 유전자인 NPT-II가 삽입되어 있음이 입증되었다 발현 정도를 비교 분석하기 위해서 NPT-ELISA 검정을 실시하였다. 삽입된 NPT II 유전자는 형질전환된 포플러의 잎, 엽병, 형성층 조직, 줄기의 목질부, 뿌리에서 발현되었으며, 발현 정도는 형질전환된 식물체의 계통에 따라서, 그리고 형진전환된 식물체의 부위에 따라서 다양하게 나타났다. pRT45에 의해서 형질전환된 Tr15 형질전환체의 경우, 늙은 잎과 엽병에서 NPT II 유전자가 가장 높은 수준으로 발현되었으며, 어린 잎과 뿌리 조직에서 가장 낮게 발현되었다. 삽입된 외래 유전자가 각 식물체간에, 각 기관에 따라서 각각 상이한 발현 정도를 나타내는 이와 같은 결과는 형질전환된 식물체에 대한 효과적인 선발과정이 요구됨을 의미함은 물론이고, 형질전환 식물체의 발달 과정에 따라서 삽입된 외래 유전자가 공간적, 시기적으로 각각 다르게 발현할 수 있다는 것을 나타낸다.

• Asian-Australasian Journal of Animal Sciences
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• 제18권5호
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• pp.626-631
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• 2005

Nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) is involved in cell apoptosis, which contributes to luteal regression and luteolysis in some species. In large domestic animals, no direct evidence for the relationship between NO and cell apoptosis in the process of corpus luteum regression is reported. The present study was conducted to investigate the localization of iNOS on porcine corpora lutea (CL) during the oestrus cycle and its relation to cell DNA fragmentation and CL regression. According to morphology, four luteal phases throughout the estrous cycle were defined as CL1, CL2, CL3 and CL4. By isoform-specific antibody against iNOS, the immunochemial staining was determined. Luteal cell DNA fragmentation was determined by flow cytometry. The results showed that no positive staining for iNOS was in CL1 and that iNOS was produced but limited to the periphery of CL2, while in the CL3, the spreading immunochemical staining was found inside the CL. No iNOS positive staining was detected in CL4. Meanwhile, DNA fragmentation increased dramatically when CL developed from CL2 to CL3 (p<0.05). In CL4, higher proportion of luteal cells still had fragmented DNA than that of luteal cells from CL1 or CL2 (p<0.05). These results indicate that iNOS expression is closely related to luteal cell apoptosis and then to luteal regression.

• 동의생리병리학회지
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• 제26권3호
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• pp.293-300
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• 2012

Inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) are important inflammatory mediators implicated in pathogenesis of inflammation and certain types of human cancers. The present study was designed to determine whether Red Ginseng (RG) could modulate $I{\kappa}B$-kinase, iNOS and COX-2 gene expression and immune responses in RAW 264.7 macrophages stimulated with lipopolysaccharide (LPS). RG extract suppressed the expression of LPS-induced $I{\kappa}B$, iNOS, COX-2, and immune responses in a dose-dependent manner. It also showed an anti-inflammatory effect by inhibiting NF-${\kappa}B$ immune response induced by LPS treatment. Inhibitory effect of RG on LPS-induced inflammation was mediated by suppressed phosphorylation of ERK, JNK and p38 through the regulation of the mitogen-activated protein kinase (MAPK) pathway leading to a decreased production of NO, iNOS, COX-2 and NF-${\kappa}B$. The results implied the role of RG as an inflammation regulator and its possible application for curing inflammatory diseases.

• 대한약침학회지
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• 제4권1호
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• pp.123-124
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• 2001

Purpose. Free radicals are implicated in the pathophysiology of aging, ischemic injury and neurodegenerative disorders. To deform]no whether Hominis Placenta extract prevents $H_2O_2$-induced apoptosis, we have performed morphological and biochemical analyses for the detection of apoptotic phenomena in the pineal tumor cell line $PGT-{\beta}$ We have also peformed cytochemical and immunocytochemical analyses for the detection of changes in nitric oxide synthase (NOS) activity and estimated the expression . of apoptotic genes using reverse transcription-polymerase chain reaction (RT-PCR) Methods. $PGT-{\beta}\;cells$ were pretreated with Hominis Placenta extracts $(0,\;10^{-2}\;{\mu}g/ml)$ for 2 hours and then exposed to $H_2O_2\;(0,\;50\;{\mu}M)$ for 3 hours. Appearance of apoptotic characteristics were monitored using 4, 6-diamidino-2-phenylindole dihydrochloride (DAPI) staining assay, terminal deoxynucleotidyl transferase-mediated dUTP-digoxigenin nick end labeling (TUNEL) assay and flow cytometric analysis. NOS activity was measured by NADPH-diaphorase cytochemistry. Expression of inducible NOS (iNOS) and nuclear factor kappa B (NF k B) was assessed via immunocytochemistry. The expression of apoptotic genes was examined by RT-PCR. Results. After 3 flours of exposure to $H_2O_2$, it was shown that $PGT-{\beta}\;cells$ treated with $H_2O_2(50\;{\mu}M)$ exhibit classical apoptotic features and increases in NOS activity and caspase-3 expression. Treatment with Hominis Placenta extract resulted in a reduced occurrence of apoptotic features. DAPI staining, TUNEL and flow cytometric assays revealed decreases in the occurrence of nuclear fragmentation and in the sub-Gl fraction in the $PGT-{\beta}\;cells$ treated with Hominis Placenta extract. Cells treated with Hominis Placenta extract also showed lower activity of NADPH-diaphorase and immunoreactivities of both iNOS and NF k B than those of $H_2O_2$-treated cells which were not treated with Hominis Placenta extract. By RT-PCR, it was shown that the level of caspase-3 mRNA was derreased In the cells treated with Hominis Placenta . extract. Conclusions. This study shows that Hominis Placenta extract prevents $H_2O_2$-induced apoptosis in $PGT-{\beta}\;cells$; inhibitions of iNOS and caspnse-3 are possible mechanisms of the protection against apoptosis.

• Journal of Acupuncture Research
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• 제25권4호
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• pp.1-9
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• 2008

Objectives : This study was to investigate the effect of low frequency electroacupuncture according to acupoints combination on NOS positive neurons in the PAG and Hippocampus of Rat with adjuvant induced Rheumatoid arthritis. Methods : Arthritis was induced by intradermal injection of FCA into base of tail. Experimental group were divided into 6 group ; Normal, Control, $ST_{36}$, $SP_9$, $ST_{36}+SP_9$, and NA. Normal group was injected with normal saline and not treated electroacupuncture. Control group was injected with FCA and not treated electroacupuncture. ST36 group was injected with FCA and treated by 2 Hz electroacupuncture at $ST_{36}$. $SP_9$ group was injected with FCA and treated by 2 Hz electroacupuncture at $SP_9$. $ST_{36}+SP_9$ group was injected with FCA and treated by 2 Hz electroacupuncture at $ST_{36}+SP_9$. NA group was injected with FCA and treated by 2 Hz electroacupuncture at non acupoint. Each groups were evaluated by the number of NOS-positive neurons in PAG and hippocampus by using an image analyzer and a microscope. Results : 1. The number of NOS-positive neurons of the NA group were significantly decreased in the CA2-3. 2. The number of NOS-positive neurons of the ST36 group were significantly decreased in the PAG and CA2-3. 3. The number of NOS-positive neurons of the SP9 group were significantly decreased in the PAG and DG 4. The number of NOS-positive neurons of the ST36+SP9 group were significantly decreased in the PAG, CA1 and DG. Conclusions : It is expected that low frequency electroacupuncture can be used a treatment of rheumatoid arthritic pain and that treatment of electroacupuncture at combined acupoints is more efficient than treatment of low frequency electroacupuncture at one acupoint.