[국내논문]현초의 항산화 활성에 의한 산화적 DNA 손상 보호효과 및 항균활성 Antimicrobial activity and protective effect of Geranium thunbergii against oxidative DNA damage via antioxidant effect원문보기
본 연구에서는 현초의 식품소재 적용과 기능성 소재의 개발을 위해 현초 에틸 아세테이트 분획물을 이용하여 활성산소종의 소거활성과 산화적 DNA 손상 보호효과 및 항균활성에 대해 검증하였다. 현초를 메탄올로 추출하여 얻어진 추출물에 대해 n-hexane, chloroform, ethyl acetate, n-butanol, water의 용매를 이용하여 순차분획을 실시하였고, 얻어진 결과물에 대하여 다양한 항산화 측정 방법을 통하여 항산화 효능을 측정한 결과 에틸 아세테이트 분획물의 경우 DPPH 라디칼 소거능, ABTS 라디칼 소거능 및 환원력에서 효과가 높게 측정 되었으며, $50{\mu}g/mL$의 농도에서 각각 80.88%, 80.12%, 28%를 저해하는 것으로 측정되었다. 이러한 항산화 효능과 함께 산화적 DNA 손상 보호효과를 검증하였고 농도별로 억제하는 경향을 나타냈다. 또한 식품 소재 및 다양한 첨가 소재로 이용하기 위하여 항균활성을 측정하였으며, 에틸 아세테이트 분획물에서 연구에 사용된 모든 균주에 대하여 저해 활성을 보였다. 이러한 활성을 가진 현초 에틸 아세테이트 분획물의 활성물질을 검증하기 위하여 phenolic compound 및 flavonoid 대조군을 이용하여 LC 분석을 하였다. 그 결과 ellagic acid와 gallic aicd가 검출 되었으며 각각 55.14 mg/g, 5.42 mg/g 측정 되었다. 이는 결과적으로 현초는 다양한 식품소재로서의 활용될 수 있으며, 본 논문은 기능성 물질로 활용을 위한 기초자료가 될 것으로 사료 된다.
본 연구에서는 현초의 식품소재 적용과 기능성 소재의 개발을 위해 현초 에틸 아세테이트 분획물을 이용하여 활성산소종의 소거활성과 산화적 DNA 손상 보호효과 및 항균활성에 대해 검증하였다. 현초를 메탄올로 추출하여 얻어진 추출물에 대해 n-hexane, chloroform, ethyl acetate, n-butanol, water의 용매를 이용하여 순차분획을 실시하였고, 얻어진 결과물에 대하여 다양한 항산화 측정 방법을 통하여 항산화 효능을 측정한 결과 에틸 아세테이트 분획물의 경우 DPPH 라디칼 소거능, ABTS 라디칼 소거능 및 환원력에서 효과가 높게 측정 되었으며, $50{\mu}g/mL$의 농도에서 각각 80.88%, 80.12%, 28%를 저해하는 것으로 측정되었다. 이러한 항산화 효능과 함께 산화적 DNA 손상 보호효과를 검증하였고 농도별로 억제하는 경향을 나타냈다. 또한 식품 소재 및 다양한 첨가 소재로 이용하기 위하여 항균활성을 측정하였으며, 에틸 아세테이트 분획물에서 연구에 사용된 모든 균주에 대하여 저해 활성을 보였다. 이러한 활성을 가진 현초 에틸 아세테이트 분획물의 활성물질을 검증하기 위하여 phenolic compound 및 flavonoid 대조군을 이용하여 LC 분석을 하였다. 그 결과 ellagic acid와 gallic aicd가 검출 되었으며 각각 55.14 mg/g, 5.42 mg/g 측정 되었다. 이는 결과적으로 현초는 다양한 식품소재로서의 활용될 수 있으며, 본 논문은 기능성 물질로 활용을 위한 기초자료가 될 것으로 사료 된다.
This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further ...
This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents. And then, its antioxidant activity was measured using various assays including total phenolic content and protection against oxidative DNA damage, and antimicrobial activities were examined using minimum inhibiting concentration (MIC) test, and paper disc method. In addition, high-performance liquid chromatography was performed to analyze the major chemical components of ethyl acetate fraction. The G. thunbergii fraction with ethyl acetate exhibited higher antioxidant and antimicrobial activities than the other fractions. The results showed that G. thunbergii ethyl acetate fraction at $50{\mu}g/mL$ had strong DPPH and ABTS radical scavenging activities of 80.88% and 80.12%, respectively. In addition, the ethyl acetate fraction protected DNA from the oxidative damage induced by ferrous ion and hydroxyl radicals and showed high antimicrobial activity with diameter of inhibition zones ranging from 13.33 to 15.67 mm. High-performance liquid chromatography analysis revealed the major phenolic compounds of G. thunbergii to be ellagic acid and gallic acid. These results suggest that G. thunbergii might protect DNA against oxidative stress induced by reactive oxygen species and can be utilized as a natural source of antioxidant and antimicrobial agent in the food industry.
This study aimed to investigate the various biological activities of Geranium thunbergii such as antimicrobial activity and protective effect against oxidative damage. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents. And then, its antioxidant activity was measured using various assays including total phenolic content and protection against oxidative DNA damage, and antimicrobial activities were examined using minimum inhibiting concentration (MIC) test, and paper disc method. In addition, high-performance liquid chromatography was performed to analyze the major chemical components of ethyl acetate fraction. The G. thunbergii fraction with ethyl acetate exhibited higher antioxidant and antimicrobial activities than the other fractions. The results showed that G. thunbergii ethyl acetate fraction at $50{\mu}g/mL$ had strong DPPH and ABTS radical scavenging activities of 80.88% and 80.12%, respectively. In addition, the ethyl acetate fraction protected DNA from the oxidative damage induced by ferrous ion and hydroxyl radicals and showed high antimicrobial activity with diameter of inhibition zones ranging from 13.33 to 15.67 mm. High-performance liquid chromatography analysis revealed the major phenolic compounds of G. thunbergii to be ellagic acid and gallic acid. These results suggest that G. thunbergii might protect DNA against oxidative stress induced by reactive oxygen species and can be utilized as a natural source of antioxidant and antimicrobial agent in the food industry.
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문제 정의
Therefore, the purpose of this study was to investigate the antioxidant and antimicrobial activities of G. thunbergii. To evaluate its antioxidant and antimicrobial activities, we first performed methanol extraction; this methanol extract was further partitioned using various solvents.
가설 설정
4)ND, not detected.
5)A-GMeans of the different superscripts within same column are significantly different at p<0.05.
6)a-dMeans of the different superscripts within same column are significantly different at p<0.05.
6)a-dMeans of the different superscripts within same row are significantly different at p<0.05.
대상 데이터
All analyses were conducted in triplicate. Statistical comparisons were carried out using SPSS 18.
Ascorbic acid, 2,2-diphenyl-1-picrylhydrazyl (DPPH), Folin-Ciocalteu’s phenol reagent, sodium carbonate, dimethyl sulfoxide (DMSO), potassium ferricyanide, butylated hydroxy anisole (BHA), trolox, trichloroacetic acid, ferric chloride and potassium persulfate were purchased from Sigma-Aldrich Chemical Co. (St. Louis, MO, USA).
The G. thunbergii was purchased from Omniherb of Gyeongsan (Gyeongbuk province, Korea). The samples were pulverized to 80 mesh size using Sung Chang Machine (ACM10INCH, Namyangju, Korea) and kept at -20℃ until assayed.
To determine antimicrobial activity, Bacillus subtilis (KCTC 1666), Bacillus cereus (KCTC 1012), Staphylococcus epidemidis (KCTC 1917), Staphylococcus aureus (KCTC 1916), Listeria monocytogenes (KCTC 3569), Escherichia coli (KCTC2643) were used. These microorganisms were kept frozen at -80℃ in broth containing glycerol (40%, v/v).
데이터처리
All analyses were conducted in triplicate. Statistical comparisons were carried out using SPSS 18.0 statistical software (SPSS, Chicago, IL, USA) and significance was determined by one-way ANOVA followed by Duncan multiple range test for multiple comparisons. Means with different letters in groups one significantly different (p<0.
이론/모형
ABTS radical cation decolorization assay was performed using the Re method (21). 7.
Antimicrobial activity of solvent-partitioned fraction was performed using the paper disc method (15). The fractions were dissolved in DMSO.
Reducing power assay was performed using the Oyaizu method (22). One microliters of the sample was added to 1 mL of potassium ferricyanide (1%, w/v) and the reaction mixture was incubated in water bath at 50℃ for 20 min.
The MIC of ethyl acetate fraction from G. thunbergii methanol extract was measured using paper disc method. The concentrations of ethyl acetate fraction were from 0.
The TPC of the extract was measured using the Folin-Denis method (19). 0.
The antioxidant activity of G. thunbergii was measured using DPPH radical scavenging assay (20). Fifty microliters of the sample was added to 100 μL of 0.
성능/효과
(30) have reported the protective effect of extract of Crataegus pinnatifida pollen against DNA damage response to oxidative stress. According to the results, C. pinnatifida pollen showed antioxidant activity, protective effect against DNA damage, and their main phenolic compound was examined. Further, Abbas et al.
6 mg/mL. All analyses were conducted under same conditions as paper disc method, and MIC was defined as the lowest concentration of ethyl acetate fraction required for inhibition of bacteria. All analyses were conducted in triplicate for each extract.
The DPPH scavenging activity of G. thunbergii solventpartitioned fractions at 50 μg/mL concentration was as following order ethyl acetate fraction > butanol fraction > chloroform fraction > water fraction > n-hexane fraction, with the following values 80.88%, 73.48%, 26.24%, 13.76%, and 13.43%, respectively.
(6) showed that the antioxidant activity of Rhizoma homalomenae extract according to total polyphenolic content increases as also shown by this study. The correlation values were estimated as DPPH/ABTS radical scavenging activity and reducing power with correlation coefficients of r2=0.9804, r2=0.9828, and r2=0.9875, respectively (data not shown).
The yield of solvent-partitioned fractions followed the order: water fraction > butanol fraction > n-hexane fraction > ethyl acetate fraction> chloroform fraction and was found to be 49.52%, 18.16%,14.51%, 6.94%, and 4.35%, respectively.
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