본 연구는 Symphoricarpos albus(S. albus) 추출물과 발효 추출물을 대상으로 zebrafish 배아를 이용하여 천연 화장품 소재로서의 유효성 평가 및 꼬리지느러미 재생력을 비교 평가하였다. 이를 위한 S. albus 추출물의 항산화 활성은 10-200μg/mL 농도에서 DPPH radical scavenging, FRAP activity, ABTS+ radical scavenging을 진행하였으며, 모두 농도 의존적인 radical 소거 활성을 보이고, S. albus 잎 추출물에 서 가장 높을 항산화 활성을 나타냈다. Zebrafish는 현재 각광받고 있는 실험 대체 동물로서 수많은 화장품 연구에 활용되고 있으며, 본 연구는 zebrafish 배아를 채취하여 응고율, 부화율, 심장 독성을 평가하였다. 그 결과 발효 추출물의 경우 100μg/mL 이상의 농도에서는 독성을 나타내는 것을 확인하였다. 재생 효능 을 평가하기 위해 zebrafish 꼬리지느러미를 절단하였고, 3일 동안 상처 회복력을 관찰하였다. 그 결과 72 시간 부터 S. albus 잎 추출물 200μg/mL에서 대조군 대비 17%의 재생 효과를 나타내었다. 이러한 결과 는 S. albus가 피부 개선용 항산화 및 재생을 위한 천연소재로서 활용 가능한 것으로 사료된다.
This study evaluated the risk of single and combined exposure to microplastics in zebrafish (Danio rerio) through biomarkers, such as survival rate, excretion rate, and histological alterations of organ systems. The experimental groups were the control (Cont.), single microplastics exposure group (MPs, 1.83%/fish total weight (g)), the copper group (Cu, 21.6 μg L-1), and a group with combined exposure to MPs and copper (MPs*Cu). The experiment was conducted with individual exposure (7 days) for MP excretion rate analysis and group exposure (14 days) for biomarker analysis. The daily excretion rate of MPs tended to decrease in a time-dependent manner. The copper concentration in the body was not significantly different between single and combined copper exposure. The degeneration of mucous cells in the skin, capillary dilation of the gill lamella, increased intestinal mucous, hepatocyte hypertrophy, and the degeneration of glomeruli and renal tubules were observed in all exposure groups. These histological alterations showed the highest tendency in the MPs*Cu group. In this study, the changes in biomarkers were attributed to the single effect of copper or the combined effect of copper and MPs rather than being solely influenced by MPs.
커피 부산물을 이용하여 대체실험동물 모델인 제브라피쉬 배아 독성 및 미백 효능에 대한 실험 을 진행하였다. 커피 부산물 추출물을 처리한 배아 독성 실험의 결과 24, 48, 72hpf에서 125ppm 농도에서 는 각각 3, 3, 5%로 응고율을 나타냈다. 배아의 부화율은 최고 농도인 125ppm에서 73%를 나타냈다. 제브 라피쉬 치어의 심장 박동수 실험에서 72hpf 후 심박수가 125ppm 농도에서 153회/60s’로 확인되었다. 음 성대조군은 148회/60s’으로 대조군의 비해 심박수의 변화가 크지 않았으며, 낮은 독성을 나타냈다. 또한 미백효능을 평가한 결과 커피 부산물 추출물의 농도가 증가할수록 멜라닌 형성이 저해되는 것으로 나타났 다. 본 연구 결과를 통해 천연 유래 부산물 소재가 화장품 원료로 활용할 수 있다는 가능성을 제안하며, 천 연 부산물의 부가가치를 높이는 연구 예시로서 화장품 산업에 활용되기를 기대한다.
Norflurazon is widely used on agricultural lands and has a high potential to pollute water sources. However, its effects on fish have not been fully elucidated. The purpose of our study was to determine whether norflurazon adversely affects the developmental stage of zebrafish, which are frequently used as a model system to evaluate the environmental impact of pollutants. Norflurazon interfered with the hatching of zebrafish embryos and induced several sublethal deformities including body length reduction, increased yolk sac volume, and enlargement of the pericardial region. We further examined the cardiotoxicity of norflurazon in the flk1:eGFP transgenic zebrafish line. The vascular network, mainly in the brain region, was significantly disrupted in norflurazon-exposed zebrafish. In addition, due to the failure of cardiac looping, norflurazon-exposed zebrafish had an abnormal cardiac structure. These developmental abnormalities were related to the apoptotic process triggered by norflurazon. Overall, the present study demonstrated the non-target toxicity of norflurazon by analyzing the hazardous effects of norflurazon on developing zebrafish.
Mecoprop-p, a chlorophenoxy herbicide, has been widely used since the 1980s. Due to its high water solubility, it could be detected in the aquatic environment, as it has already been detected in the surface water or groundwater in several countries. The toxicity of other chlorophenoxy herbicides has been reported; however, there are few studies on the toxicity of mecoprop-p, one of the chlorophenoxy herbicides, on aquatic organisms. Here, we investigated the toxic effects of mecoprop-p using zebrafish. After mecoprop-p exposure, we observed that the zebrafish larvae eyes did not form normally, heart edema was generated, and the body length was shortened. The number of cells undergoing apoptosis also increased in the anterior part including head, heart, and yolk sac of the mecoprop-p-treated zebrafish compared to the untreated controls. Moreover, cardiovascular structures, including the heart and aortic arches, were also malformed after exposure to mecoprop-p. Therefore, our results suggest that mecoprop-p could cause abnormal development in zebrafish larvae and there is also a high possibility that mecoprop-p would be toxic to other aquatic organisms.
다양한 동물 모델이 인간 질병, 의약품의 효능 및 작용 메커니즘을 연구하는 데 사용되고 있다. Zebrafish(Danio rerio)는 여러 가지 장점이 있어 인간 질병에 대한 중개 연구의 모델로 점점 더 폭넓게 활용되고 있다. 본 논문은 Pubmed, Google Scholar, Scopus에서 2020년 12월까지 최근 10년간 zebrafish 모델, 천연물(한약), in vivo 스크리닝의 키워드를 사용하여 저널에 게재된 논문을 검토하여 필 요한 정보를 얻었다. 이 리뷰에서 우리는 천연물(한약) 연구에 대한 다양한 제브라피쉬 질병 모델의 최 근 경향에 대해 논의하였다. 특히, 암, 안질환, 혈관 질환, 당뇨병 및 합병증, 피부질환에 중점을 두었고, zebrafish 배아를 사용하여 이들 질병에 대한 의약품의 분자 작용 메커니즘에 관해 언급하였다. Zebrafish는 실험실에서 임상 연구까지의 격차를 줄이는 데 중추적 역할을 할 수 있는 중요한 동물 모 델이다. Zebrafish는 의약품이나 화장품 개발, 질병의 병인론을 이해하기 위해 사용되고, 이로 인해 생의 학 연구에서 설치류의 사용을 줄이는 데 크게 기여하고 있다.
In this study, the potential toxicity of isoprocarb was demonstrated using zebrafish embryos. We treated isoprocarb (0, 29, and 58 mg/L) to the zebrafish embryos for 72 h then, we estimated morphological changes and apoptotic cell numbers. The increasing extent of apoptosis from the anterior to posterior region of developing zebrafish larvae was correlated with toxicity in the overall development process, including growth and normal organ formation. The appearance of abnormalities in the isoprocarb-treated groups in comparison to normal developing zebrafish larvae was verified using quantitative image analysis based on ImageJ software program. The vascular system comprising a complex interconnection of blood vessels was visualized in vessel-fluorescent transgenic zebrafish (fli1:eGFP). The main vasculature was malformed on isoprocarb treatment, and this was also related to cardiac defects. Taken together, normal embryonic development in zebrafish was interrupted owing to the acute toxicity of isoprocarb.
Dimethachlor is a synthetic herbicide, belonging to the chloroacetanilide group, that inhibits the undesirable growth of weeds via the suppression of very longchain fatty acid synthesis. Although dimethachlor has been shown to run off from agricultural fields into aquatic ecosystems, the toxicity of dimethachlor on aquatic invertebrates and vertebrates is unknown. In our study, we assessed the toxicity of dimethachlor on developing zebrafish embryos by analyzing viability, hatching ability, and phenotypic changes. Embryonic viability decreased from 48 h post-fertilization (hpf) at the highest concentration of dimethachlor. Decreased hatching ratio, shortened body length, and pathological changes in the eye, heart, and yolk sac were observed at sub-lethal concentrations. Additionally, dimethachlor increased the number of apoptotic cells and level of reactive oxygen species 120 hpf. Our results indicate that dimethachlor may act as an anti-developmental toxicant when accumulated in an aquatic environment.
This study investigated the regenerative and healing effect of spirulina on corneal epithelial injury by acid burn in zebrafish. Corneal epithelium of adult zebrafish was evaluated at 0, 12, 24, 48 h after acid burn injury with or without immersion with 0.02 and 0.04 mg/mL of spirulina water extract. The pathological changes of corneal epithelium were assessed by light microscopy. Gene and protein expressions of proliferating cell nuclear antigen (PCNA), matrix metalloproteinase (MMP) 9 and MMP13 were determined by immunohistochemistry or real-time PCR. Corneal epithelium was denuded totally after acid burn and gradually regenerated. PCNA-positive cells significantly increased in spirulina treated group. MMP9 and MMP13 mRNA transcripts were significantly decreased in spirulina treated groups. In conclusion, spirulina has regenerative and healing properties by increased keratocyte proliferation, and inhibited extracellular matrix degradation during the regeneration process of corneal epithelial injury by acid burn in zebrafish.