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潘祥龙,郝二伟,谢金玲,杜正彩,侯小涛,邓家刚.基于斑马鱼模型的海洋中药厚藤安全性评价[J].广西科学,2019,26(5):560-568. [点击复制]
- PAN Xianglong,HAO Erwei,XIE Jinling,DU Zhengcai,HOU Xiaotao,DENG Jiagang.Safety evaluation of marine Chinese medicine Ipomoea pes-caprae (Linn.) Sweet based on zebrafish model[J].Guangxi Sciences,2019,26(5):560-568. [点击复制]
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| 基于斑马鱼模型的海洋中药厚藤安全性评价 |
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潘祥龙1,2,3, 郝二伟1,2,3, 谢金玲1,2,3, 杜正彩1,2,3, 侯小涛1,2,3, 邓家刚1,2,3
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| (1.广西中医药大学, 广西中药药效研究重点实验室, 广西南宁 530001;2.广西农作物废弃物功能成分研究协同创新中心, 广西南宁 530200;3.中-加斑马鱼中药筛选联合实验室, 广西南宁 530200) |
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| 摘要: |
| 采用模式生物斑马鱼模型评价海洋中药厚藤水提物（Ipomoea pes-caprae（Linn.） Sweet water extract，IWE）及醇提物（Ipomoea pes-caprae（Linn.） Sweet alcohol extract，IAE）的安全性。采用受精后3 dpf （Day post fertilization） AB系斑马鱼进行急性毒性评价，将受精后10 hpf （Hours post fertilization）斑马鱼用于胚胎发育毒性及心脏毒性评价；将受精后3 dpf的gz15Tg/+（AB）肝脏荧光蛋白转基因斑马鱼用于肝脏毒性评价。结果表明，急性毒性实验中厚藤水提物半数致死浓度（LC50）为2.84 mg/mL，10%致死浓度（LC10）为1.46 mg/mL；厚藤醇提物LC50为1.32 mg/mL，LC10为0.76 mg/mL。胚胎发育毒性实验中厚藤水提物最大非致死浓度（MNLC）组及LC10组干预24 h对斑马鱼胚胎未见尾部自主运动现象且发育迟缓，斑马鱼发育至72 hpf出现心脏和软黄囊肿大现象；厚藤醇提物LC10组与空白对照组比较出现心包水肿，血瘀，观察中各组未见胚胎发育畸形现象。心脏毒性实验中空白对照组斑马鱼胚胎心跳节律规则、搏动强有力；厚藤水提物及醇提物在24 hpf，48 hpf，72 hpf时间点1/3MNLC组、MNLC组及LC10组分别与空白对照组比较，均能明显减慢斑马鱼胚胎的心率且呈现剂量依赖性（P<0.01）；厚藤水提物及醇提物1/3MNLC组、MNLC组及LC10组分别与空白对照组比较，均能明显增加斑马鱼胚胎的SV-BA间距（P<0.01）。肝脏毒性实验中厚藤水提物中各给药组与空白对照组比较，肝脏荧光面积均有减少趋势，组间比较无差异性；厚藤醇提物中1/3MNLC组、MNLC组及LC10组与空白对照组比较，肝脏荧光面积均有增加趋势，但组间比较无差异性，各给药组斑马鱼的丙氨酸氨基转移酶（ALT）、天冬氨酸转氨酶（AST）水平与空白对照组比较均无显著差异。本实验中厚藤水提物的LC50显著高于醇提物。高剂量LC10组均对斑马鱼胚胎发育产生一定毒性作用。厚藤水提物及醇提物随着给药剂量增加，明显减慢斑马鱼胚胎的心率，产生一定心脏毒性作用。厚藤水提物及醇提物均未引起明显的肝脏毒性。 |
| 关键词: 厚藤 斑马鱼 急性毒性 胚胎毒性 心脏毒性 肝脏毒性 |
| DOI：10.13656/j.cnki.gxkx.20191024.009 |
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| 基金项目:广西科技计划项目（17-259-20，桂科AB17195019，桂科合15104001-11和桂科AD17195025）资助。 |
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| Safety evaluation of marine Chinese medicine Ipomoea pes-caprae (Linn.) Sweet based on zebrafish model |
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PAN Xianglong1,2,3, HAO Erwei1,2,3, XIE Jinling1,2,3, DU Zhengcai1,2,3, HOU Xiaotao1,2,3, DENG Jiagang1,2,3
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| (1.Guangxi Key Laboratory for Pharmacodynamics Research of Traditional Chinese Medicine, Guangxi University of Chinese Medicine, Nanning, Guangxi, 530001, China;2.Collaborative Innovation Center of Study on Functional Ingredients of Agricultural Residues, Nanning, Guangxi, 530200, China;3.Sino-Canada Joint Zebrafish Lab for Chinese Herbal Drug Screening, Nanning, Guangxi, 530200, China) |
| Abstract: |
| The model organism zebra fish model was used to evaluate the safety of the marine Chinese medicine of Ipomoea pes-caprae (Linn.) Sweet water extract (IWE) and Ipomoea pes-caprae (Linn.) Sweet alcohol extract (IAE).Three dpf (Day post fertilization) AB zebra fish was used for acute toxicity evaluation,and 10 hpf (Hours post fertilization) zebra fish was used for embryonic developmental toxicity and cardio-toxicity evaluation.After fertilization at 3 dpf,gz15Tg/+ (AB) liver fluorescent protein transgenic zebra fish was used for liver toxicity evaluation.The results showed that in the acute toxicity experiment,the LC50 of the IWE was 2.84 mg/mL,the LC10 was 1.46 mg/mL,the LC50 of the IAE was 1.32 mg/mL,and the LC10 was 0.76 mg/mL.In the embryonic developmental toxicity test,the IWE MNLC group and the LC10 group were treated for 24 h.There was no active movement of the tail in the zebra fish embryo and the development was slow.The zebra fish developed to 72 hpf and the heart and soft yellow cysts appeared.Compared with the blank control group,LC10 group of the IAE showed pericardial edema and blood stasis.No abnormalities in embryonic development were observed in each group.In the cardio-toxicity experiment,the zebra fish embryos had strong heartbeat rhythm rules and strong beats in the blank control group.The IWE and IAE at 24 hpf,48 hpf,72 hpf,1/3 MNLC group,MNLC group and LC10 group were compared with the blank control group,respectively,the heart rate of zebra fish embryos was significantly slowed down in a dose-dependent manner (P<0.01).The IWE and IAE 1/3 MNLC group,MNLC group and LC10 group were compared with the blank control group,both can significantly increased the SV-BA spacing of zebra fish embryos (P<0.01).In the liver toxicity test,each administration group was compared with the blank control group,liver fluorescence area all showed a trend of decrease,but there was no difference between the groups.The IAE 1/3 MNLC group,MNLC group and LC10,compared with the blank control group,the liver fluorescence area increased,but there was no difference between the groups.The ALT and AST levels of zebra fish in each group were not significantly different from those in the blank control group.In this experiment,the LC50 of IWE was significantly higher than that of IAE.High-dose LC10 group had certain toxic effects on zebra fish embryo development.The IWE and IAE significantly slowed the heart rate of zebra fish embryos with the increase of dose,which produced certain cardio-toxic effects.Both the IWE and IAE did not cause significant liver toxicity. |
| Key words: Ipomoea pes-caprae (Linn.) Sweet zebrafish acute toxicity embryotoxicity cardiotoxicity hepatotoxicity |
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