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目的:研究冰片对斑马鱼胚胎发育的影响。方法:以斑马鱼胚胎为模型,二甲基亚砜为助溶剂,制备冰片溶液(33.6、67.1、134.3、268.5、537.0、1 074.0μmol/L,即冰片①、②、③、④、⑤、⑥组),并且设空白对照组,以受精后小时(hpf)为时间段,对3hpf胚胎进行暴露染毒处理,每24 h更换1次冰片溶液,分别在8、24、48、72、96 hpf时间点以显微镜观察班马鱼胚胎发育形态,测定24 hpf斑马鱼胚胎自主抽动次数、48 hpf心率、孵化率、畸形率和死亡率。实时荧光定量聚合酶链反应法检测斑马鱼胚胎Sepn1基因表达的情况。结果:与空白对照组比较,冰片④、⑤、⑥组斑马鱼胚胎48 hpf出现脊柱弯曲、心囊水肿、卵黄囊水肿,游泳异常的毒性症状;冰片②、③、④、⑤、⑥组斑马鱼胚胎24 hpf自动抽动次数减少;冰片④、⑤、⑥组斑马鱼胚胎24 hpf心率减少;在48 hpf时,冰片①、②、③、④、⑤、⑥组斑马鱼胚胎孵化率降低;在72 hpf时,冰片②、③、④、⑤、⑥组斑马鱼胚胎孵化率降低;在96 hpf时,冰片⑤、⑥组斑马鱼胚胎孵化率降低;冰片④、⑤、⑥组斑马鱼胚胎96 hpf畸形率增加;24 hpf后,冰片④、⑤、⑥组斑马鱼胚胎死亡率降低;冰片④组Sepn1 mRNA表达增强,差异有统计学意义(P<0.01或P<0.05)。结论:高剂量冰片对斑马鱼胚胎发育有明显影响,且呈现一定的时间依赖性。建议婴幼儿、孕妇和哺乳期的妇女长时间或大剂量使用含冰片的药物均应谨慎。
Objective: To study the effects of borneol on zebrafish embryo development. Methods: The zebrafish embryo was used as a model and dimethyl sulfoxide as cosolvent to prepare borneol solution (33.6,67.1,134.3,268.5,537.0,1 074.0μmol / L, namely borneol ①, ②, ③, ④, ⑤, ⑥ group), and set the blank control group to fertilization after hours (hpf) for the time period, exposure to 3hpf embryo exposure treatment, a replacement ice every 24 h solution, respectively, at 8,24,48,72,96 hpf time point observation of the morphological characteristics of the embryos of the development of the genus Bambu, determination of 24h hpf zebrafish embryo autonomic twitches, 48hpf heart rate, hatchability, deformity and mortality. Detection of Sepn1 Gene Expression in Zebrafish Embryos by Real-time Fluorescent Polymerase Chain Reaction. Results: Compared with the blank control group, the 48 hpf of the embryos of borneus ④, ⑤ and ⑥ group showed spinal flexion, edema of the sac, yolk sac edema and abnormal swimming symptoms. The borneol ②, ③, ④, ⑤, ⑥ groups of zebra The frequency of 24 hpf auto-tibia was decreased in fish embryos; the heart rate of 24 hpf in the groups of ¢ ó, ¢ ¢ and ¢ ö decreased significantly At 72 hpf, the hatching rate of zebrafish embryos decreased in ¢ ò, ¢, ¢, ¢, ¢ ¢ and ¢ ¢ ¢ õ groups. At 96 hpf, (P <0.01 or P <0.05). After 24 hpf, the mortality of zebrafish embryos decreased in ¢ ó, ¢ ¢, and ¢ ö groups. The expression of Sepnl mRNA increased in ¢ ö ④ group (P <0.01 or P <0.05). Conclusion: High doses of borneol significantly affect embryonic development of zebrafish and exhibit a time-dependent manner. It is recommended that babies, pregnant women and lactating women should be cautious about the use of borneol-containing drugs for prolonged or high doses.