香加皮水提取物对斑马鱼幼鱼肝脏毒性的初步研究

Effects of waterextract from cortex periplocae on hepatotoxicity of zebrafish

目的研究香加皮水提取物对斑马鱼幼鱼肝脏的毒性作用。方法将受精后发育4天的斑马鱼幼鱼暴露于不同浓度的香加皮水提取物中,24小时后,统计幼鱼的死亡率,计算量毒曲线;选取低于LC_(10)的三个药物暴露组和空白组,以幼鱼肝脏形态和面积变化、转基因幼鱼肝脏荧光面积和荧光强度以及幼鱼肝脏细胞凋亡、超氧化物歧化酶(superoxide dismutase,SOD)、谷胱甘肽过氧化酶(glutathione peroxidase,GSH-Px)、丙氨酸转氨酶(alanine aminotransferase,ALT)、天门冬氨酸转氨酶(aspartic transaminase,AST)活性为毒性评价指标。结果香加皮水提取物对幼鱼的量-毒回归曲线y=-1.0843+0.0014x(R^2=0.9524,r=0.97590,P<0.001),LC_(10)为845.9286μg/m L;与空白组相比,幼鱼肝脏形态异常、透明度降低,肝脏面积随药物暴露浓度增加而剂量依赖性增加(P<0.05);转基因幼鱼肝脏荧光面积随暴露浓度增加而增大,但在高浓度组减小,荧光强度随暴露浓度的增加而减小(P<0.05);高浓度暴露组幼鱼肝脏区域显示了细胞凋亡;药物暴露组幼鱼SOD、GSH-Px活力显著下降,ALT、AST活力显著上升(P<0.05)。结论香加皮水提取物对斑马鱼幼鱼的肝脏毒性可能是通过破坏其体内氧化应激平衡,进而诱导肝脏细胞凋亡实现的。

Objective To study the toxic effect of waterextract from cortex periplocae on liver of zebrafish larval. Methods Zebrafish embryos of 4 days post-fertilization were exposed to 12-well plates with different concentrations of waterextract from cortex periplocae for 24 hours, and the mortality rate was calculated and the toxic curve was calculated. Three drug exposed groups less than LC10 and the blank group were selected. The changes of liver morphology and area, fluorescence area and fluorescence intensity of liver, apoptosis of liver cell, superoxide dismutase, glutathione peroxidase, alanine aminotransferase, aspartic transaminase were used as indicator of toxicity assessment. Results The LC10 of waterextract from cortex periplocae was 845. 9286 μg/ mL,the mortality rate was dependent with the concen-tration, concentration-mortality curve was y= -1. 0843＋0. 0014x （R2 =0. 9524,P〈0. 001）,and correlation coefficient was 0. 97590（P〈0. 001）. Compared with the control group, morphological abnormalities was more and transparency was decreased in liver, and liver area was increased with drug exposure concentration （P〈0. 05）; The fluorescence area was increased with the exposure concentration increased, but in the high concentration group, the fluorescence intensity was decreased with the exposure concentration increased （P〈0. 05）; The apoptosis was appeared in high concentration exposed group. The activity of SOD, GSH-Px was significantly decreased while the activity of AST, ALT was significantly increased. Conclusion Waterextract from cortex periplocae can induce hepatotoxicity in zebrafish larvae, and the mechanism may relate to destroy the balance of oxidative stress and induce apoptosis in liver cells.