苗药了哥王提取物对斑马鱼的毒性作用

Study on the toxicity of the extract from Miao medicine Wikstroemia indica to zebrafish

目的研究苗药了哥王提取物对斑马鱼的毒性作用。方法以斑马鱼胚胎模型为对象,经10、20、40μg/mL了哥王提取物暴露处理后,检测并记录1 min内自主抽动次数、10 s内心率及其畸形、死亡发生情况;以斑马鱼模型为对象,经10~100μg/mL了哥王提取物暴露处理24、48、72 h后,计算各时间点了哥王提取物对斑马鱼的半数致死浓度(LC50);经低、中、高质量浓度(27、37、51μg/mL)了哥王提取物暴露处理后,观察斑马鱼肝脏表型和细胞凋亡、脂质沉积情况,检测其肝组织中丙氨酸转氨酶(ALT)、天冬氨酸转氨酶(AST)、乳酸脱氢酶(LDH)活性。结果与空白组比较,了哥王提取物10、20、40μg/mL组斑马鱼胚胎的自主抽动次数、畸形率(10μg/mL组除外)、死亡率均显著升高,胚胎心率(10、20μg/mL组除外)均显著降低(P<0.05);了哥王提取物干预24、48、72 h对斑马鱼的LC50分别为39.850、28.300、21.490μg/mL;了哥王提取物低、中、高质量浓度组斑马鱼的肝脏区域透明度降低、形态膨大,细胞凋亡和脂质沉积均有所增加,肝组织中ALT、AST、LDH活性均较空白组显著升高(P<0.05)。结论了哥王提取物对斑马鱼胚胎具有发育毒性,对斑马鱼具有肝损伤毒性。

OBJECTIVE To study the toxicity of the extract from Miao medicine Wikstroemia indica to zebrafish.METHODS Zebrafish embryo model was used as the object,after exposure to W.indica extract(10,20,40μg/mL),the number of spontaneous twitching within 1 min,heart rate within 10 s,the occurrence of malformation and death were detected and recorded.Zebrafish was used as the object,after exposure to W.indica extract(10-100μg/mL)for 24,48 and 72 h,and the median lethal concentration(LC50)of W.indica extract to zebrafish at different time points were calculated.After exposure to low,medium and high concentration(27,37,51μg/mL)of W.indica extract,the liver phenotype,hepatocyte apoptosis and lipid deposition of zebrafish were observed,and the activities of alanine aminotransferase(ALT),aspartate aminotransferase(AST)and lactate dehydrogenase(LDH)in liver tissue were detected.RESULTS Compared with blank group,the number of spontaneous twitching,malformation rate(except for 10μg/mL group)and mortality of embryos increased significantly in 10,20,40μg/mL groups of W.indica extract,and the heart rate(except for 10,20μg/mL group)of embryos decreased significantly(P<0.05).LC50of W.indica extract to zebrafish at 24,48 and 72 h were 39.850,28.300 and 21.490μg/m L,respectively.After drug treatment,the transparency of liver area of zebrafish in low,medium and high concentration groups of W.indica extract reduced and their shape were enlarged;apoptosis and lipid deposition increased;the activities of ALT,AST and LDH in liver tissue were significantly increased(P<0.05).CONCLUSIONS Miao medicine W.indica extract had developmental toxicity to zebrafish embryos and hepatotoxicity to zebrafish.