Oxidative stress has been reported to be one of causes of neuritis. This study examined antioxidative activities of methanol extracts of six amphibian species known to be medicinal animals (Rana catesbeiana, R. coreana, R. rugosa, R. dybowskii, R. nigromaculata, and Hyla japonica) and investigated t...
Oxidative stress has been reported to be one of causes of neuritis. This study examined antioxidative activities of methanol extracts of six amphibian species known to be medicinal animals (Rana catesbeiana, R. coreana, R. rugosa, R. dybowskii, R. nigromaculata, and Hyla japonica) and investigated their effects of inhibiting nitric oxide (NO) production and cytotoxicity on the murine macrophage RAW264.7 cells. As inflammation is closely associated with reactive oxygen species, assays on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, xanthine oxidase inhibitory activity, superoxide anion radical scavenging activity and NO scavenging activity of the extracts of the six species were performed to investigate their antioxidative activity. The results obtained were as follows; All extracts showed antioxidative activity, and the activity of R. dybowskii was the highest in comparison among those. Anti-inflammatory effects of the extracts were also examined, the five extracts except that of R. rugosa did not show cytotoxicity for RAW264.7 cells at the maximal concentration ($1,000{\mu}g\;mL^{-1}$). Selectivity index, meaning NO scavenging activity compared to cytotoxicity, showed the highest level in the extract of R. dybowskii. These results will be very useful basic data for future studies on prevention and treatment of human diseases to understand the biological roles of amphibian extracts throughout the antioxidative or anti-inflammatory pathways.
Oxidative stress has been reported to be one of causes of neuritis. This study examined antioxidative activities of methanol extracts of six amphibian species known to be medicinal animals (Rana catesbeiana, R. coreana, R. rugosa, R. dybowskii, R. nigromaculata, and Hyla japonica) and investigated their effects of inhibiting nitric oxide (NO) production and cytotoxicity on the murine macrophage RAW264.7 cells. As inflammation is closely associated with reactive oxygen species, assays on 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activity, xanthine oxidase inhibitory activity, superoxide anion radical scavenging activity and NO scavenging activity of the extracts of the six species were performed to investigate their antioxidative activity. The results obtained were as follows; All extracts showed antioxidative activity, and the activity of R. dybowskii was the highest in comparison among those. Anti-inflammatory effects of the extracts were also examined, the five extracts except that of R. rugosa did not show cytotoxicity for RAW264.7 cells at the maximal concentration ($1,000{\mu}g\;mL^{-1}$). Selectivity index, meaning NO scavenging activity compared to cytotoxicity, showed the highest level in the extract of R. dybowskii. These results will be very useful basic data for future studies on prevention and treatment of human diseases to understand the biological roles of amphibian extracts throughout the antioxidative or anti-inflammatory pathways.
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문제 정의
96) and its efficiency was observed to be the highest among the extracts. These results of this study are considered to be important basic data for studies on antioxidants and anti-inflammatory components through extraction of active ingredients from amphibians and on isolation and mechanism of active components to prevent or to treat diseases. In the future, in vitro and in vivo biological functions of sequential fractions of amphibians are needed to be examined and additional researches on possibility of an industrial use of EtOH or spirit extracts are also considered to be necessary.
제안 방법
In traditional oriental medicine, it is used for nephropathy, diuresis, nutrition and flatulence, dried powder in warm honey water or boiled amphibian bodies by itself is eaten as a special efficient medicine of pulmonary tuberculosis, roborant and asthma. This study was conducted to provide necessary data for researches on antioxidants and anti-inflammatory materials and on their isolation and mechanism to prevent and cure diseases by investigating biological activity such as antioxidant activity and anti-inflammatory effect of methanol (MeOH) extracts of six amphibian species collected in Korea, which has been rarely studied and has been reported to be used as a fork remedy.
대상 데이터
Murine macrophage cell line RAW264.7 was obtained from Korean Cell Line Bank (Seoul, Republic of Korea) and was incubated at 37℃ with 5% CO2 conditions using Dulbecco’s modified Eagle’s medium (DMEM) including 100 units mL-1 penicillin-streptomycin and 10% fetal bovine serum (FBS).
Six amphibian species (R. catesbeiana, R. coreana, R. rugosa, R. dybowskii, R. nigromaculata, and H. japonica) were used for this study obtained from American Bullfrog Capture Operation Division (Jeongeup, Jeollabuk-do, Republic of Korea) and Frog Village (Muju, Jeollabuk-do, Republic of Korea). After lyophilization of animal specimens, samples were subsequently grinded, deposited on 500 mL 80% MeOH, and extracted three times by using a sonicator.
데이터처리
The results were presented with a mean and standard deviation and a statistical significance was analyzed with Student’s t-test.
이론/모형
To examine antioxidant activity of each sample, the Blois (1958) method measuring radical scavenging effect with DPPH (Sigma, USA) was used. DPPH solution was made by dissolving around 2 mg DPPH in 15 mL EtOH.
성능/효과
According to the results, MeOH extracts of R. catesbeiana, R. coreana, R. dybowskii, R. nigromaculata, and H. japonica did not show cytotoxicity to the maximal concentration, 1,000μg mL-1 and TC50 of R. rugosa showing 50% cytotoxicity was observed to be 333.6μg mL-1.
NO scavenging activity of the MeOH extracts of the six species of Amphibia was measured with the amount of nitrite by using SNP forming NO. At 2.5 mg mL-1 each sample showed 48~15% scavenging activity and the scavenging effect of R. dybowskii, R. nigromaculata and H. japonica was found to be good by recording 48.1, 42.5 and 35.1%, respectively while that of R. coreana, R. catesbeiana and R. rugosa was relatively low by recording 26.7, 20.2 and 15.6%, respectively. Like previous studies on antioxidant activity of medicinal insect extracts revealing that the extracts of Anomala albopilosa, Sympetrum eroticum and Anax parthenope had high antioxidant activity (Kim et al.
In addition, as R. dybowskii did not trigger cytotoxicity to 1,000μg mL-1 and its selectivity index was also the highest (6.96) and its efficiency was observed to be the highest among the extracts.
In conclusion, all MeOH extracts of the six species had antioxidant activity, and in comparison among those the activity of R. dybowskii was found to be the best by considering DPPH radical scavenging activity, xanthine oxidase inhibitory activity and NO scavenging activity. In addition, as R.
6μg mL-1. Selectivity index meaning NO scavenging activity compared to cytotoxicity was the highest in R. dybowskii by recording 6.96 and those of R. rugosa, R. coreana, R. catesbeiana, R. nigromaculata and H. japonica recorded 4.63, 3.46, 3.29, 2.92 and 2.51, respectively (Table 2, Fig. 2).
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