• 제목/요약/키워드: spectrofluorometer

검색결과 23건 처리시간 0.017초

UVB로 산화적 손상을 유도한 피부섬유아세포에 Saponaria 추출물의 보호효과 (Protective Effect of Saponaria Extract Against UVB-Damage in Skin Fibroblasts)

  • 김보애
    • 한국응용과학기술학회지
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    • 제39권5호
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    • pp.644-651
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    • 2022
  • 피부는 인체를 구성하는 가장 큰 장기로 생체 내부를 보호한다. 자외선은 피부에 광노화와 산화적 손상을 비롯한 다양한 염증반응을 일으킨다. 본 연구의 목적은 섬유아세포에서 UVB를 조사하여 Saponaria 추출물의 보호 효과를 조사하는 것이다. 본 연구에서는 UVB에 의한 세포독성과 산화적 세포사멸, NO 및 PGE2 생성에 대한 보호활성을 나타내는 Saponaria의 유효성을 평가하였다. HS68 세포를 UVB(120mJ/cm2)에 조사하고 100, 200, 400 ㎍/mL의 다양한 농도로 Saponaria 추출물로 24시간 동안 처리하였으며, 자외선 B에 의해 생성된 세포 내 활성 산소 종(ROS)은 DCF-DA 염색 후 분광 형광계를 사용하여 검출하였다. 또한 지질 과산화는 배양 배지로 분비되는 8-이소프로스탄의 수준을 측정하여 분석하였다. 그 결과 Saponaria 추출물이 UVB에 의한 세포독성을 효과적으로 억제하였다. 산화적 세포 손상은 UVB로 유도된 HS68 섬유아세포에서 PGE2를 매개하였고, 이는 사포나리아 추출물 처리에 의하여 유의하게 억제되었다. 또한, 이들 추출물의 보호 효과는 농도 의존적으로 세포내 ROS 생성 및 지질 과산화 억제에 의해 매개되는 것으로 평가되었다. 이러한 결과는 Saponaria 추출물이 자외선 B에 의한 산화적 스트레스로 매개한 피부 손상을 억제하여 세포 보호효과를 나타내므로 항노화 기능성 소재로 활용될 수 있을 것으로 사료된다.

황백(黃柏)의 berberine이 DNA의 기능조절에 미치는 영향에 관한 형광이방성 연구 (Fluorescence Anisotropy Study on the Effect of Phellodendri Cortex's Berberine on Regulation of the Function of DNA)

  • 이성경;한효상;허성호
    • 대한본초학회지
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    • 제33권5호
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    • pp.105-110
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    • 2018
  • Objectives : We tried to observe the fluorescence anisotropy and intensity of ethidium ion in the intercalating binding interaction between DNA and ethidium ions in the presence of berberine, and then tried to explain the effect of berberine on the intercalating interaction of ethidium ion with DNA. Methods : DNA(calf thymus DNA), berberine and ethidium bromide(EtBr) were purchased from Sigma-Aldrich Co. Proper amount of each compound was dissolved in 20 mM sodium phosphate buffer(pH 7.0) containing 100 mM of NaCl to prepare stock solutions. Collections of the fluorescence anisotropy and intensity data were performed on JASCO FP-8300 spectrofluorometer equipped with a polarizer and a Peltier temperature controller. The excitation of ethidium ion was done at 550 nm and the emission data were collected at 600 nm. For Stern-Volmer plot, the fluorescence data were collected at $18^{\circ}C$ and $30^{\circ}C$. Results : According to the results of this research, the weak competitive binding pattern between ethidium ion and berberine appeared in binding with DNA at low ratio of DNA to ethidium ion. But at high ratio of DNA to ethidium ion, this weak competition disappeared. Instead, berberine might bind to DNA by intercalating way. In other words, berberine could de-intercalate ethidium ion from DNA at low concentration of DNA relative to ethidium ion, but could not at high concentration of DNA relative to ethidium ion. In addition, the mechanism of fluorescence quenching of ethidium ion could also proceed differently, depending on the ratio of the amount of DNA to that of ethidium ion. Conclusions : The effect of berberine on the DNA-ethidium ion intercalating interaction could work differently, depending on the relative ratio of the amount of DNA to that of ethidium ion. This study also showed that fluorescence anisotropy analysis is very useful method to obtain detailed information for investigation of the complex binding interactions. In order to fully understand the mechanism of action of the pharmacological effect by berberine, studies on the effect of berberine on the action of proteins such as various enzymes closely related to berberine-induced medicinal effects should be continued.

정상인에서 혈중 연과 zinc protoporphyrin과의 상관관계 및 HPLC 와 Hematofluorometer로 측정한 zinc protoporphyrin량간의 비교 (The Relationship between Zinc protoporphyrin and Lead levels in Normal Adults' Blood And Comparison of Zinc protoporphyrin values by High Performance Liquid Chromatograph and Hematofluorometer)

  • 김강윤;김현욱
    • 한국산업보건학회지
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    • 제3권2호
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    • pp.141-151
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    • 1993
  • Blood samples obtained from 200 adults who had visited the "S" general hospital were analyzed to compare the zinc protoporphyrin (ZPP) levels quantified by high performance liquid chromatograph (HPLC) and by hematofluorometer (HF) to investigate the methodological difference if any and the relationship between the levels of blood lead and ZPP among no-lead exposed adults. Also investigated were the distribution of ZPP and protoporphyrin IX (PPIX) concentrations, the establishment of normal levels of blood ZPP and blood lead, and the contribution of age and sex factors to these values. These subjects had no previous occupational exposure to lead. The results obtained were as follows : 1. The mean values of blood lead for male and female subjects were $9.46{\pm}2.44{\mu}g/dl$ and $8.09{\pm}2.17{\mu}g/dl$, respectively. The difference observed in the mean concentrations between male and female subjects was statistically very significant. 2. The mean values of blood ZPP by HPLC for male and female subjects were $15.94{\pm}4.55{\mu}g/dl$ and $22.26{\pm}6.61{\mu}g/dl$, respectively. The difference observed in the mean concentrations between male and female subjects was statistically not significant. The mean values of blood PPIX by HPLC for male and female subjects were $2.51{\pm}1.78{\mu}g/dl$ and $2.81{\pm}1.56{\mu}g/dl$, respectively. The difference observed in the mean concentrations between male and female subjects was statistically not significant. 3. The mean values of blood ZPP by HF for male and female subjects were $28.44{\pm}7.11{\mu}g/dl$ and $37.77{\pm}8.04{\mu}g/dl$, respectively. The difference observed in the mean concentrations between male and female subjects was statistically very significant. 4. No statistically significant correlation was found between the levels of blood ZPP and blood lead. 5. The ratio of ZPP and protoporphyrin IX (PPIX) concentration to erythrocyte protoporphyrin (EP, EP=ZPP+PPIX) concentration was 87.4% and 12.6%, respectively. 6. A statistically very significant correlation was found between the ZPP concentrations determined by HPLC and the values by HF (r=0.7565). The ZPP concentraitons quantified by HF were 1.75 times as high as the values obtained by HPLC. 7. The blood ZPP concentrations quantified by HPLC, HF, and spectrofluorometer (SF) from the blood samples obtained from 14 lead-exposed workers and from 16 no-lead exposed adults showed wide variations. The ZPP concentrations by HF were the highest followed by the levels obtained by SF and by HPLC. In the exposed group, no statistically significant difference was found among three methods of quantifying blood ZPP levels. In the no-lead exposed group, however, statistically significant difference was observed among these methods. The ZPP concentrations by HF were about twice as high as those of by HPLC or by SF. Among three methods of quantifying blood ZPP (HPLC, SF and HF), the results revealed significant difference. Therefore it is suggested that objective methods of quantifying blood ZPP and a system of correcting different ZPP levels be developed by the ministry of Labor.

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