Acute Toxicity and Oxidative Stress Effects of Aerial Stem and Leaf Extract of Notopterygium incisum on Zebrafish(Danio rerio) Embryos
LIU Kang-Kang1, LI Yong-Juan1,2,*, HUANG Jin-Qiang1, WANG Qi1, GUO Zhi-Jia1, CHEN Xue-Mei1, WANG Xiao1
1 College of Animal Science and Technology, Gansu Agricultural University, Lanzhou 730070, China; 2 College of Science, Gansu Agricultural University, Lanzhou 730070, China
Abstract：Notopterygium incisum, as an endemic traditional Chinese herbal resource in northwestern China, its root extracts have shown unique potential in the field of aquatic pathogen prevention and control. However, systematic studies on the effects of stem and leaf extracts on aquatic organisms have not been conducted domestically or internationally, resulting in a lack of necessary safety data to support the development and utilization of these stems and leaves. To realize the rational exploitation of N. incisum stem and leaf resources, this study prepared an ethanol-water extract of the stems and leaves via ultrasonic extraction. Using zebrafish(Danio rerio) embryos as the test organism, a 96 h acute exposure experiment was carried out to detect changes in antioxidant enzyme activities and the expression levels of related genes, thereby investigating the acute toxicity and oxidative stress effects of the extract at low(1/5 LC50, 5.8 mg/L), medium(2/5 LC50, 11.6 mg/L), and high(4/5 LC50, 23.2 mg/L) concentrations on zebrafish embryos. Results showed that the stem and leaf extract of N. incisum inhibited the hatching of zebrafish embryos, with a 96 h median lethal concentration(96 h-LC50) of 29 mg/L. Under low and medium concentrations, the teratogenicity was low, and no significant effects on body length and heart rate. In contrast, high concentrations caused malformations in zebrafish embryos and larvae, such as trunk cysts, delayed yolk sac absorption, and pericardial edema. Antioxidant enzyme activities and the expression of related genes also exhibited concentration dependence: The activities of SOD, CAT, and GST were significantly decreased at low, medium, and high concentrations in a concentration dependent manner, and the MDA content was significantly higher at the high concentration than that at the low and medium concentrations(P<0.05). Specifically, low concentrations mainly induced the expression of antioxidant genes such as mn-sod and gstp-2; medium concentrations significantly activated core antioxidant pathway genes including ucp-2 and cat; high concentrations inhibited the expression of cox-1 and cat, but induced nqo-1, indicating that high concentrations might trigger more complex oxidative stress or toxic responses. Furthermore, the expression levels of cell apoptosis-related genes(bax and bcl-2) were significantly increased at medium concentrations, suggesting that medium concentrations were more likely to induce molecular responses associated with zebrafish embryo apoptosis. This study provides fundamental data for the development and utilization of abandoned herbal resources(stems and leaves of N. incisum) and the research of green aquatic drugs.
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