• 동의생리병리학회지
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• 제34권1호
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• pp.7-13
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• 2020

Sanyeoleumja (SY) is the traditional Korean medicinal prescription for the treatment of inflammatory diseases of eyes. In this study, the anti-inflammatory effects of SY water extract were investigated. To measure the anti-inflammatory effects of SY, we examined the productions of inflammatory factor including nitric oxide (NO), prostaglandin E2 (PGE₂), inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), interleukin-6 (IL-6), interleukin-1β (IL-1β) in lipopolysaccharide (LPS)-induced RAW 264.7 cells. SY inhibited NO and PGE₂ production in a dose dependent manner and decreased the protein and mRNA expression of iNOS and COX-2. Also, SY decreased the mRNA expression of interleukin-6 (IL-6) and interleukin-1β (IL-1β). In conclusion, SY downregulated LPS-induced inflammatory factor productions, which could be a clinical basis for inflammatory diseases.

• 한국식품과학회지
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• 제49권3호
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• pp.343-348
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• 2017

상백피 및 상지 에탄올 추출물의 항염증 효과를 확인하기 위하여 RAW 264.7 세포를 사용하여 시험하였다. 상백피, 상지 추출물의 활성산소종, 산화질소(II), 프로스타글란딘 $E_2$, 인터류킨-6, 종양괴사인자-${\alpha}$ 등의 생성능과 COX-2, iNOS mRNA 발현을 확인하였다. 산화질소(II) 생성량은 두 추출물 모두 유의하게 비슷한 정도로 감소시켰으나, 활성산소종 생성에서는 상백피 추출물이 상지 추출물 보다 억제 효과가 높게 나타났다. 프로스타글란딘 $E_2$와 인터류킨-6 생성에서는 상지 추출물의 억제 효과가 강하게 나타났으며, COX-2와 iNOS mRNA 발현량 또한 상지 추출물의 상백피에 비해 유의하게 감소시킴을 확인하였다. 본 연구에서 확인한 결과로 상백피, 상지 추출물 모두 염증 매개인자 발현 억제를 통해 항염증 효과를 가짐을 알 수 있었으며, 상지 추출물이 상백피 추출물 보다 염증 완화 효과가 강한 것으로 나타났다. 향후에는 상백피 및 상지 추출물의 주요 활성 성분들의 차이와, 염증완화 메커니즘을 통한 관련 질환에 효과가 있는지 정확한 작용메커니즘 확인 등의 분석실험이 필요할 것으로 생각된다.

• Journal of Microbiology and Biotechnology
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• 제24권11호
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• pp.1464-1472
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• 2014

The microbial community structures of two continuous stirred tank reactors digesting turkey manure with pine wood shavings as well as chicken and swine manure were investigated. The reactor fed with chicken/swine wastes displayed the highest organic acids concentration (up to 15.2 g/l) and ammonia concentration (up to 3.7 g/l ammonium nitrogen) and generated a higher biogas yield (up to $366ml/g_{VS}$) compared with the reactor supplied with turkey wastes (1.5-1.8 g/l of organic acids and 1.6-1.7 g/l of ammonium levels; biogas yield was up to $195ml/g_{VS}$). The microbial community diversity was assessed using both sequencing and profiling terminal restriction fragment length polymorphisms of 16S rRNA genes. Additionally, methanogens were analyzed using methyl coenzyme M reductase alpha subunit (mcrA) genes. The bacterial community was dominated by members of unclassified Clostridiales with the prevalence of specific clostridial phylotypes in each reactor, indicating the effect of the substrate type on the community structure. Of the methanogenic archaea, methanogens of the genus Methanosarcina were found in high proportions in both reactors with specific methanosarcinas in each reactor, whereas the strict hydrogenotrophic methanogens of Methanoculleus sp. were found at significant levels only in the reactor fed with chicken/swine manure (based on the analyses of 16S rRNA gene). This suggests that among methanogenic archaea, Methanosarcina species which have different metabolic capabilities, including aceticlastic and hydrogenotrophic methanogenesis, were mainly involved in anaerobic digestion of turkey wastes.

• Archives of Pharmacal Research
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• 제27권7호
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• pp.769-775
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• 2004

Hepatic ischemia and reperfusion (l/R) predisposes the liver to secondary stresses such as endotoxemia, possibly via dysregulation of the hepatic microcirculation secondary to an imbalanced regulation of the vascular stress genes. In this study, the effect of hepatic I/R on the hepatic vasoregulatory gene expression in response to endotoxin was determined. Rats were subjected to 90 min of hepatic ischemia and 6 h of reperfusion. Lipopolysaccharide (LPS, 1 mg/kg) was injected intraperitoneally after reperfusion. Plasma and liver samples were obtained 6 h after reperfusion for serum aminotransferase assays and RT-PCR analysis of the mRNA for the genes of interest： endothelin-1 (ET-1), its receptors $ET_A$ and $ET_B$, endothelial nitric oxide synthase (eNOS), inducible nitric oxide synthase (iNOS), heme oxygenase-1 (HO-1), cyciooxygenase-2 (COX-2), and tumor necrosis factor-a (TNF-${\alpha}$). The activities of serum aminotransferases were significantly increased in the I/R group. This increase was markedly potentiated by LPS treatment. The ET-1 mRNA was increased by LPS alone, and this increase was significantly greater in both the I/R alone and I/R ＋ LPS groups compared to the sham. There were no significant differences in ETA receptor mRNA levels among any of the experimental groups. $ET_B$ mRNA was increased by both LPS alone and I/R alone, with no significant difference between the I/R alone and I/R ＋ LPS groups. The eN OS and HO-1 transcripts were increased by I/R alone and further increased by I/R ＋ LPS. The iNOS mRNA levels were increased by I/R alone, but increased significantly more by both LPS alone and I/R ＋ LPS compared to I/R alone. The TNF-${\alpha}$ mRNA levels showed no change with I/R alone, but were increased by both LPS alone and I/R ＋ LPS. The COX-2 expression was increased significantly by I/R alone and significantly more by I/R ＋ LPS. Taken collectively, significantly greater induction of the vasodilator genes over the constriction forces was observed with I/R ＋ LPS. These results may partly explain the increased susceptibility of ischemic livers to injury as a result of endotoxemia.

• 약학회지
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• 제47권5호
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• pp.311-318
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• 2003

Eurya emarginata (Thunb.) Makino (Theaceae) is distributed in coastal areas of island. The leaves of Eurya are used in the traditional medicine of the coastal areas of jeju island with the aim of diuresis or to treat ulcers. Nevertheless, there are few reports on the biological activity and constituents of E. emarginata. In this study, we investigated the pharmacological activity of the solvent extracts of E. emarginata on the several inflammatory markers (TNF-$\alpha$, IL-1$\beta$, IL-6, NO, iNOS and COX-2). Also we examined the antioxidizing effect of the solvent extracts by determination of DPPH radical-scavenging activity. Among the solvent fractions, EtOAc and BuOH extracts showed potent radical scavenging activity (RC$_{50}$=10.9 and 12.7 respectively). The subtractions of EF 5-4-6-3-2 and BF 1 potentially inhibited the mRNA expression of pro-inflammatory cytokines (IL-1$\beta$, IL-6 and TNF-$\alpha$) at the concentration of 100 $\mu\textrm{g}$/mι. Also the fractions inhibited the mRNA expression of pro-inflammatory cytokines (IL-1$\beta$, IL-6 and TNF-$\alpha$) and protein expression of iNOS and COX-2 at the concentration of 100 $\mu\textrm{g}$/mι. And then, the inhibition of iNOS was correlated with the decrease of nitrite level. These results suggest that E. emarginata may have anti-inflammatory activity through the inhibition of pro-inflammatory cytokines, iNOS and COX-2.2.

• 대한약침학회지
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• 제6권3호
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• pp.5-14
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• 2003

Objective : Recently, Herbal-acupuncture therapeutics has been used for the treatment of inflammatory diseases such as rheumatoid arthritis. Especially, we have been interested in chemical mediators concerned with inflammation such as prostaglandin, cytokine, nitric oxide. The purpose of this study is investigated that the effect of Scutellariae Radix-acupuncture solution in lipopolysaccharide-stimulated RAW 264.7 macrophages, performed several expeimental items : those are prostaglandin E. nitric oxide and cyclooxygenase-2. Method : The cytotoxicity of Scutellariae Radix-acupuncture solution in RAW 264.7 macrophages were measured by MTT-based cytotoxicity assay. In order to observe cyclooxygenase-2 mRNA expression in lipopolysaccharide-stimulated RAW 264.7 macrophages, RT-PCR was used. Prostaglandin $E_2$ formation and nitric oxide production was measured by competitive enzyme immunoassay and Griess assay. Results : 1. The MTT assay demonstrated that cytotoxic effect of Scutellariae Radix-acupuncture solution in RAW 264.7 macrophages were not appeared before concentration of 5mg/mL 2. Scutellariae Radix-acupuncture solution inhibited cyclooxygenase-2 mRNA expression in lipopolysaccharide-treated RAW 264.7 macrophages. 3. Scutellariae Radix-acupuncture solution inhibited nitric oxide production in lipopolysaccharide-treated RAW 264.7 macrophages. 4. Scutellariae Radix-acupuncture solution inhibited prostaglandin $E_2$ formation in lipopolysaccharide-treated RAW 264.7 macrophages. Conclusion : On the basis of these results, It was shown that Scutellariae Radix-acupuncture solution is significantly able to inhibit the production of $PGE_2$ and NO, as well as COX-2 mRNA expression. Our results may provide new mechanism by which Scutellariae Radix-acupuncture solution accounts for its beneficial effect on accelerating wound healing and anti-inflammation.

• 생약학회지
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• 제52권2호
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• pp.92-98
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• 2021

Gryllus bimaculatus extract (GbE) have reported that anti-inflammatory activity by suppression of pro-inflammatory cytokines. However, the effects of fermented Gryllus bimaculatus extract (FGbE) have not yet been investigated. In this study, we evaluated the anti-inflammatory and anti-wrinkle effect of the fermented Gryllus bimaculatus extracts using Bacillus subtilis (JB PMB-18) in RAW264.7 cells. Both GbE and FGbE exerted no cytotoxic effects until 1000 ㎍/mL concentration. FGbE decreased NO production and decreased iNOS and COX-2 mRNA levels in a concentration-dependent manner. In addition, the protein production of inflammatory cytokines TNF-α, IL-1β and IL-6 was effectively reduced compared to the GbE. Inhibitory activities of elastase and collagenase associated with skin wrinkle improvement were measured to be 45% and 69%, respectively, at a concentration of 500 ㎍/mL in FGbE. From these results, FGbE can be used as a health functional food and skin functional cosmetic materials for preventing inflammatory diseases because it has excellent anti-inflammatory and anti-wrinkle effects.

• 자연과학논문집
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• 제9권1호
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• pp.19-24
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• 1997

PCR 방법을 이용하여 K11 RNA 중합효소를 coding하는 Klebsiella phage gene 1을 cloning 하였고 lac 전사촉진제 조절 하에 발현시켰다. K11 RNA 중합효소는 DAEA-sephacel과 Affigel blue column chromatographies를 사용하는 상용 방법으로 분리하였다. DAEA-sephacel의 0.2-0.3 M $NH_4Cl$ 분획에서 K11 RNA 중합효소의 활성을 보였고, 다음 단계의 Affigel blue column에서 SDS-polyacryl amide gel 상의 단일 band로 분리되었다. K11 RNA 중합효소는 T7 그룹 phage RNA 중합효소로 다른 T7 그룹phage RNA 중합효소와 많은 상동성을 보인다. (대장균 phage T7, T3과 Salmonella tyhimurium phage SP6 RNA 중합효소). 이미 우리는 T7과 SP6 전사촉진제 변이체를 제조한 바 있고 T7과 SP6 RNA 중합효소의 전사촉진제 특이성을 연구한 바 있다 (이상수와 강창원, 1993). K11 RNA 중합효소의 전사촉진제 특이성을 알아보기 위해 SP6 전사촉진제 변이체를 사용하여 in vitro K11 RNA 중합효소의 활성을 측정하였다. 이 변이체 중 K11 전사촉진제와 가장 유사한 것이 가장 높은 K11 RNA 중합효소 활성을 보였다.

• 동의생리병리학회지
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• 제17권4호
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• pp.1065-1074
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• 2003

In order to evaluate the antiallergic effects of Kamiyukgunja-tang (KYGJT), studies were done. We measured the cytotoxic activity for lung fibroblast cell, cytokines transcript expression, production of INF-γ, IL-10, IL-4, GM-CSF, IL-1 β, TNF-α. IL-5 proliferation of B cell in anti-CD40mAb plus r1L-4 stimulated murine splenic B cells. The results were obtained as follows : 1. KYGJT was not showed cytotoxicity in the fibroblast lung cell. 2. KYGJT increased the gene synthesis of INF-γ, IL-10, GM-CSF(m-RNA). 3. KYGJT decreased the gene synthesis of IL-1β, IL-4, TNF-α, IL-5(m-RNA). 4. KYGJT decreased the appearance of TNF-α significantly. 5. KYGJT decreased the appearance of IgE significantly. 6. KYGJT decreased the proliferation of B cell significantly. 7. KYGJT decreased the appearance of Histamin Release Production significantly. The facts above prove that KYGJT is effective against the allergy. Thus. I think that we should study on this continuously

• Archives of Pharmacal Research
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• 제21권3호
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• pp.291-297
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• 1998

Cop protein has been overexpressed in Escherichia coli using a T7 RNA polymerase system. Purification to apparent homogeneity was achieved by the sequential chromatography on ion exchange, affinity chromatography, and reverse phase high performance liquid chromatography system. The molecular weight of the purified Cop was estimated as 6.1 kDa by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). But the molecular mass of the native state Cop was shown to be 19 kDa by an analytical high performance size exclusion chromatography, suggesting a trimer-like structure in 50 mM Tris-HCI buffer (pH 7.5) containing 100 mM NaCl. Cop protein Was calculated to contain $39.1% {\alpha}-helix, 16.8% {\beta}-sheet$, 17.4% turn, and 26.8% random structure. The DNA binding property of Cop protein expressed in E. coli Was preserved during the expression and purification process. The isoelectric point of Cop was determined to be 9.0. The results of amino acid composition analysis and N-terminal amino acid sequencing of Cop showed that it has the same amino acid composition and N-terminal amino acid sequence as those deduced from its DNA sequence analysis, except for the partial removal of N-terminal methionine residue by methionyl-aminopeptidase in E. coli.