• 대한한의학방제학회지
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• 제28권4호
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• pp.339-349
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• 2020

Objectives : Wiryeong-tang (WRT) is a traditional herbal medicine used to treat kidney-related diseases. However, the anti-inflammatory and anti-gastritis effect of Wiryeong-tang was not well known. Therefore, we experimented to confirmed the anti-inflammatory and anti-gastritis effects of Wiryeong-tang. Methods : The RAW 264.7 cells were pre treated with Wiryeong-tang mix soft extract (WRT-mse; 50, 100 and 200 ㎍/mL) for 1 hrs, and then incubated with lipopolysaccharide (LPS; 500 ng/mL). Cell viability was measured by the MTT method, and nitric oxide (NO) was measured with griess reagent. In addition, pro-inflammatory cytokines were measured by enzyme-linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR). For anti-gastritis effect in vivo, acute gastritis was induced using 150 mM HCl/60% ethanol used ICR mice. WRT-mse (133 mg/kg) was pre treated for 3 days and then treated with 150 mM HCl/60% ethanol 1 hrs later. Then gastritis was observed and inflammatory cytokines in the gastric tissue was measured. Results : The 8 marker components of the WRT-mse were determined by simultaneous analysis using HPLC. WRT-mse was not toxic and inhibited pro-inflammatory cytokines such as IL-1β, IL-6 and TNF-α at NO production, protein and mRNA levels. Also, it was confirmed that WRT-mse improved bleeding and edema in gastritis, and suppresses inflammatory cytokines. Conclusion : In summary, our results suggest that the treatment of the WRT-mse reduced and improved the 150 mM HCl/60% ethanol induced acute gastritis and the inflammation caused by LPS stimulation in RAW 264.7 cells. Therefore, this study may provide useful drug or clinical evidence for WRT-mse to prevent inflammation.

• 대한한의학회지
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• 제37권1호
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• pp.21-33
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• 2016

This study aimed to investigate the stability and biological activities of BHSST decoction depending on the preservation temperature and periods. Methods: BHSST decoction was preserved at room temperatures (R/T, $23{\pm}1^{\circ}C$) or refrigeration ($4^{\circ}C$) for 0, 30, 60 and 90 days. To evaluate the stability of BHSST decoction, pH and sugar content were estimated. In addition, high-performance liquid chromatography (HPLC) analysis was performed to determine marker compounds of BHSST decoction. To evaluate anti-inflammatory effect, nitric oxide (NO) and prostaglandin $E_2$ ($PGE_2$) productions were measured in LPS-stimulated RAW 264.7 macrophages. Antioxidant activity was examined using the assays for 3-ethyl-benzothiazoline-6-sulfonic acid (ABTS) and 1-1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities. Results: There was no change in pH and sugar content depending on the preservation temperature and periods of BHSST decoction. Among the major components of BHSST, contents of liquiritin, baicalein and wogonin was reduced time-dependently both at R/T and $4^{\circ}C$. Inhibitory effects of BHSST decoction on NO and PGE2 productions were slightly decreased in a time-dependent manner by 90 days of preservation. In addition, BHSST decoction maintained ABTS and DPPH radical scavenging activities by 60 days while significantly reducing the activities in 90 days of preservation at R/T. By contrast, BHSST decoction had no significant change of ABTS and DPPH radical scavenging activities by 90 days at $4^{\circ}C$. Conclusions: Our results suggest that the stability and efficacy of BHSST decoction are maintained for 60 days at $4^{\circ}C$ rather than R/T.

• BMB Reports
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• 제38권1호
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• pp.58-64
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• 2005

Myo-inositol monophosphate phosphatase (IMPP) is a key enzyme in the phosphoinositide cell-signaling system. This study found that incubating the IMPP from a porcine brain with pyridoxal-5'-phosphate (PLP) resulted in a time-dependent enzymatic inactivation. Spectral evidence showed that the inactivation proceeds via the formation of a Schiff's base with the amino groups of the enzyme. After the sodium borohydride reduction of the inactivated enzyme, it was observed that 1.8 mol phosphopyridoxyl residues per mole of the enzyme dimer were incorporated. The substrate, myo-inositol-1-phosphate, protected the enzyme against inactivation by PLP. After tryptic digestion of the enzyme modified with PLP, a radioactive peptide absorbing at 210 nm was isolated by reverse-phase HPLC. Amino acid sequencing of the peptide identified a portion of the PLP-binding site as being the region containing the sequence L-Q-V-S-Q-Q-E-D-I-T-X, where X indicates that phenylthiohydantoin amino acid could not be assigned. However, the result of amino acid composition of the peptide indicated that the missing residue could be designated as a phosphopyridoxyl lysine. This suggests that the catalytic function of IMPP is modulated by the binding of PLP to a specific lysyl residue at or near its substrate-binding site of the protein.

• Journal of Plant Biotechnology
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• 제31권3호
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• pp.231-237
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• 2004

Oxycarotenoids는 녹색식물, 곰팡이, 효모, 버섯 및 세균 등이 만들어 내는 황색, 적색 또는 자색의 polyene계 색소로 분자내에 산소를 함유하며 생체내에서 중요한 역할을 담당하고 있다. 본 실험에서는 Oxycarotenoids의 생합성 경로상에 존재하는 $\beta$-carotene hydroxylase 유전자 (Chyb)가 재조합된 Ti-plasmid (pGCHYB)를 A. tumerfacience GV3101에 의해 Arabidopsis thaliana (cv. Columbia)에 형질전환하였다. 50 mg/L hygromycin 함유한 MS 배지에서 선발된 개체를 이용하여 Chyb 유전자의 도입여부를 PCR로 분석한 결과, 대조구에서는 Chyb 유전자의 증폭 되지 않았으나 형질전환체에서는 증폭 산물을 확인 할 수 있었다. 또한 형질전환체의 발현여부를 RT-PCR분석한 결과 도입된 Chyb 유전자가 안정적으로 발현되었다. 형질전환체의 carotenoids를 HPLC 분석한 결과 xanthophyll cycle carotenoids (violaxanthin과 zeaxanthin)의 함량 및 $\beta$-carotene 함량은 감소되었으며, 대조구 Arabidopsis에는 생합성되지 않는 astaxanthin이 생합성되었다. 따라서 본 실험에서 육성된 형질전환체를 이용하여 oxycarotenoids 생합성 과정상의 중간대사물질의 표지, 관여된 transcript 및 metabolite 분석 등을 통해 carotenoids 대사계의 연구소재로 활용 할 수 있을 것으로 기대한다.

• Journal of Microbiology and Biotechnology
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• 제8권2호
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• pp.171-177
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• 1998

The strain A49, which produces a new type of extracellular polysaccharide was isolated from soil samples. From morphological, physiological and biochemical tests, the strain A49 was identified as a Bacillus polymyxa and named Bacillus polymyxa A49. Bacillus polymyxa A49 was found to produce a highly viscous extracellular polysaccharide when grown aerobically in a medium containing glucose as the sole source of carbon. The polysaccharide (A49 POL) showed a homogeneous pattern on gel permeation chromatography (GPC) and its molecular weight was estimated to be about 1.6 mega dalton (mDa). The FT-IR spectrum of A49-POL revealed typical characteristics of polysaccharides. As a result of investigations with HPLC and carbozole assay, A49-POL was found to consist of L-fucose, D-galactose, D-glucose, D-mannose, and D-glucuronic acid, with the molar ratio of these sugars being approximately 1:2:7:50:12. Rheological analysis of A49 POL revealed that it is pseudoplastic and has a higher apparent viscosity at dilute concentrations than does xanthan gum. The consistancy factor of A49 POL was found to be higher, and the flow index of A49 POL lower, than xanthan gum. Its apparent viscosity was comparatively unstable at various temperatures. the A49 POL showed the highest apparent viscosity at pH 3. When salts were added to A49 POL solution, the solution was compatible with up to 10% KCl, 35% NaCl, 55% $CaCl_2$, 55% $MgCl_2$, 55% $K_2HPO_4$, and 110% $Ca({NO_3})_2$, respectively.

• Journal of Microbiology and Biotechnology
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• 제14권5호
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• pp.985-990
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• 2004

Levans produced from Microbacterium laevaniformans were isolated, characterized, and fractionated by molecular weight. TLC, HPLC, and GC-MS analyses of the exopolysaccharide showed that it was a fructan-type polymer and was composed of (2,6)- and (2,1)-glycosidic linkages. $^{13}C$-NMR analysis proved that the polysaccharide was mainly a $\beta$-(2,6)-linked levan-type polysaccharide. To investigate the cytotoxicity of the acetone-precipitated levan fractions such as M1, M2, and M3, HepG2, P388D1, U937, SNU-1, and SNUC2A cell lines were screened. Among the cell lines tested, the cytotoxicity of M1- M3 fractions were detected from only SNU-1 and HepG2 cells at the dosage level of $100-800\mu\textrm{g}ml$. The M2 fraction M_r$, 80,000) at 400 $mu{g/ml}$ had the greatest cell growth inhibition (84.6%) on SNU-1, while the M1 $(M_r$, 50,000) at $800\mu\textrm{g}ml$ showed the greatest (46.32%) on HepG2. To obtain more uniform M_r$ fractions of levan, the levan was further fractionated from S1 $(M_r$ 1,000,000) to S5 $(M_r$ 10,000) using gel permeation chromatography. Again, the S1-S5 fractions had strong cytotoxicity on SNU-1 and HepG2 cell lines. The greatest inhibition effects of S4 $(M_r$ 80,000) on SNU-1 and S5 $(M_r$ 10,000) on HepG2 were shown to be 49.5% and 73.0%, respectively. The cytotoxicity of the levan fractions was more effective on SNU-1 than on HepG2. Although the relationship between the Mw and the cytotoxicity was not clear, smaller $M_r$, fractions of levan showed greater growth inhibition effect on the cancer cell lines in general. Therefore, it was indicated that a specific Mw class of levan is responsible for the effective cytotoxicity.

• Asian Journal of Atmospheric Environment
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• 제5권3호
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• pp.146-151
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• 2011

1-Nitropyrene (1-NP), 2-NP and 2-nitrofluoranthene (2-NFR) are useful markers for studying the atmospheric behaviors of polycyclic aromatic hydrocarbons (PAHs) and nitropolycyclic aromatic hydrocarbons (NPAHs). However, present methods for measuring trace levels of these compounds are lesssensitive and laborious. Here we describe several improvements to a previously reported high-performance liquid chromatography-chemiluminescence detection system that allows it to determine trace levels of 1-, 2-NPs and 2-NFR. The proposed system was equipped with a reducer column packed with Pt/Rh instead of zinc whose life-time was limited. The combination of Cosmosil MS-II (monomeric ODS) and AR-II (polymeric ODS) columns was used instead of polymeric ODS columns as the separator column to improve the separation. An ethanol mixture with acetate buffer (pH 5.5) was used in place of an acetonitrile mixture with the same buffer to activate the reducer column. The same ethanol mixture was used as the mobile phase for the clean-up column. The switching time of the column switching valve was optimized to concentrate the amino-derivatives of above NPAHs quantitatively on the concentrator column. The concentrations of bis(2,4,6-trichlorophenly) oxalate and hydrogen peroxide in the chemiluminescence reagent solution were optimized to 0.4 mM and 30 mM, respectively, to increase the sensitivity. Under the above conditions, the detection limits (S/N=3) of 1-, 2-NPs and 2-NFR were 1 fmol (0.25 pg), 10 fmol (2.5 pg) and 4 fmol (1 pg), respectively. The proposed system was effectively used to determine trace levels of 1-, 2-NPs and 2-NFR in airborne particulates collected at Noto Peninsula. The atmospheric concentrations of 1-, 2-NPs and 2-NFR were not more than sub pg $m^{-3}$ levels. They were higher in winter (January) than in summer (July). In both seasons, the concentrations were in decreasing order, [2-NFR]>[1-NP]>[2-NP].

• Applied Biological Chemistry
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• 제51권3호
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• pp.188-193
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• 2008

수삼을 수분함량 별로 건조한 다음 원형이 변하지 않는 팽화 인삼을 제조할 수 있는 최적의 압력조건을 찾아 팽화 인삼을 제조하고 각 조건 별 시료의 추출수율, 조사포닌, ginsenoside함량 변화를 관찰하였다. 팽화 처리 후 외관상의 가장 큰 변화는 갈변과 부피팽창이었다. 추출수율 측정 결과, 대조군은 37.6%, 팽화 인삼의 경우 $50.0{\sim}62.1%$의 범위로 측정되었다. 조사포닌 함량의 경우 대조군은 11.0 mg/g ginseng, 팽화 인삼은 $19.6{\sim}48.8mg/g$ ginseng의 범위로 측정되었다. 팽화 인삼에서는 홍삼 특유 사포닌인 ginsenoside-Rg3가 검출되었다. Ginsenoside-Rg3의 생성량은 팽화 압력이 증가함에 따라 유의적으로 증가하는 경향을 나타내었다. Ginsenoside-Rg3를 제외한 나머지 기본 ginsenoside와 total ginsenoside함량은 대조군에 비해 모두 증가하였지만, 팽화 압력이 증가함에 따라 그 양이 감소하는 경향을 나타내었다. 따라서 본 연구 결과 인삼을 적절한 조건에서 팽화처리하였을 경우 추출수율, 조사포닌 및 조사포닌 함량의 증진과 일부 ginsenoside의 변형을 확인할 수 있었다.

• Journal of Applied Biological Chemistry
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• 제61권2호
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• pp.109-117
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• 2018

최근까지도, 현대사회의 암 발생률은 급격하게 증가하고 있다. 인체 내부에서 내재적 또는 외재적인 요인에 의해 DNA 손상이 발생되고, 세포는 DNA 손상에 대한 방어기작을 통해 스스로를 방어한다. 또한, 비정상적인 DNA 생성 및 결손된 DNA 가닥의 복원은 노화, 암, 염증 등 다양한 질병으로부터 기인한다. 많은 연구자는 이러한 DNA 손상을 억제하기 위하여 적절한 소재 탐색에 많은 관심을 두고 있으며, 특히 합성화합물의 부작용이 알려지면서, 천연물을 기반으로 한 암 예방적 소재에 대한 연구가 많이 이루어지고 있다. 토복령은 백합과(Liliacese)에 속하는 청미래덩굴(Smilax china L.)의 근경이며, 전통적으로 해독과 종기 등의 치료제로 사용되어왔다. 하지만 토복령의 DNA 손상에 대한 억제 효과에 대한 연구는 미흡하다. 본 논문에서는 토복령의 항산화 효과 및 DNA 손상에 대한 억제 효과를 확인하고, 식물이 포함하는 phenolic 화합물의 활성과 연관 관계를 확인하고자 하였다. 항산화 효과를 확인하기 위해, DPPH 라디칼 및 ABTS 라디칼에 대한 소거 활성을 확인하였다. 토복령 추출물은 DPPH 및 ABTS 라디칼을 효과적으로 제거하였으며, 높은 환원력을 나타냈다. HPLC 분석을 통해 phenolic 화합물을 정량 및 동정하였으며, 항산화 효과와 phenolic 화합물의 연관 관계를 확인하였다. 또한, $OH^-$ 라디칼 및 $Fe^{2+}$으로 유발된 plasmid DNA 손상에 대한 방어 효과를 확인하였다. 세포 수준에서, DNA 손상에 대한 저해 효과는 산화적 스트레스로 유발된 NIH 3T3 세포의 ${\gamma]$-H2AX 및 p53 단백질 발현 저해 활성을 확인하였다. 또한, H2AX 및 p53 mRNA 수준의 저해 활성을 확인하였다. 결론적으로, 토복령 추출물의 phenolic 화합물의 항산화 효과 및 DNA 손상에 대한 억제 효과를 확인하였다.

• 대한임상검사과학회지
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• 제39권3호
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• pp.196-200
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• 2007

호모시스테인은 관상동맥질환, 뇌혈관 질환의 위험인자로 알려지고 있다. 초기의 호모시스테인 측정은 HPLC를 이용하여 일반적인 검사실에서 이용하기에는 어려움이 많았으며 현재 여러 가지 면역분석기가 도입되어 시행되고 있다. 하지만 최근 자동화학분석기를 이용한 새로운 효소법의 시약이 개발되어 이 시약의 유용성 평가를 하기위해 본 연구에서는 기존의 면역분석기와 비교분석하였다. 그 결과 정밀도 평가에서 변이계수는 0.98 - 3.23%, 2.55~4.58%를 나타내었고, 자동화학분석기(TBA 200FR와 Advia 1650)를 이용한 새로운 효소법(HBI)과 기존의 면역분석기(Advia Centaur와 Immulite 2000)간의 상관관계 평가에서 상관계수는 0.9632와 0.9625로 우수한 상관관계를 보였다. 이 결과를 토대로 호모시스테인 측정에 있어 새로운 효소법의 자동화학분석기를 이용한 방법이 제시되어 보다 간편하고 쉬운 routine analysis에 적합하리라 사료된다.