| 引用本文:韩沁1，邹迪新2，李二文1，李芝奇1，徐玥1，赵崇军1，林瑞超1.芫花水提取物对斑马鱼成鱼及SD大鼠肝毒性的相似性评价及作用机制初探[J].世界中医药,2020,(13):. |
|
| 芫花水提取物对斑马鱼成鱼及SD大鼠肝毒性的相似性评价及作用机制初探 |
| Exploration on Similar Evaluation and Mechanism of Hepatotoxicity of Water Extract from Genkwa Flos in Adult Zebrafish and SD Rats |
| 投稿时间：2020-06-10 |
| DOI：10.3969/j.issn.1673-7202.2020.13.005 |
| 中文关键词: 芫花 水提取物 斑马鱼 大鼠 肝毒性 基因表达 |
| English Keywords:Genkwa Flos Water extract Zebrafish Rat Hepatotoxicity Gene expression |
| 基金项目:国家中医药管理局国家中药标准化项目（ZYBZH-Y-YN-44）——三七等2种中药饮片标准化建设 |
|
| 摘要点击次数: 631 |
| 全文下载次数: 0 |
| 中文摘要: |
| 目的:比较芫花水提取物对斑马鱼成鱼及SD大鼠肝毒性作用的相似性，并初步探讨芫花肝毒性作用机制。方法:选取斑马鱼成鱼320条，分为32组，每组10条（雌:雄=1:1），设置高、中、低剂量组和空白对照组，暴露处理96 h。SD大鼠24只，分为4组，每组6只，设置高、中、低剂量组和空白对照组，按照2.5 mL/200 g体质量每日灌胃给药1次，连续7 d。在实验终点，将斑马鱼成鱼和大鼠处死并获取肝脏组织。通过病理组织切片观察其肝脏形态，以SOD、SDH、MDA、NOX、TG、ALT和AST等生化指标为毒性评价指标。在此基础上通过实时荧光定量PCR测定斑马鱼成鱼肝脏相关基因的表达情况。结果:病理组织切片显示，芫花处理能够引起斑马鱼成鱼肝脏细胞变形、空泡，甚至凋亡，同时,芫花能够引起大鼠肝脏细胞排列松散、空泡、形态皱缩、界限不清晰，细胞核形态成不规则状并伴有出血现象.生化测定结果表明,芫花处理组斑马鱼成鱼肝脏中SOD、SDH及NADH的酶活力均显著降低（P<0.05），而MDA含量上升（P<0.01）；给药组大鼠肝脏中SOD、SDH的活力均显著下降（P<0.05），而MDA、TG含量升高（P<0.01），此外，低剂量给药组大鼠ALT活力显著性升高（P<0.05），所有给药组中AST活力均有所升高（P<0.01）。qRT-PCR结果显示，斑马鱼肝脏组织中线粒体功能相关基因mtatp6，mtco1，mtnd1，pklr表达水平下调(P<0.01),氧化应激相关基因gstp2，nqo1，sod1的mRNA表达水平下调(P<0.01)，内质网应激相关基因包括atf4a，ern1，hsp90b1的mRNA表达水平上调(P<0.01)，自噬相关基因beclin-1和ATG3表达水平下调(P<0.01)。结论:芫花水提取物对斑马鱼成鱼及SD大鼠肝脏均产生毒性作用，其作用机制可能为芫花能够诱导斑马鱼肝脏组织中出现氧化应激损伤、线粒体功能障碍、内质网应激和细胞自噬，进而诱导肝脏细胞凋亡实现的。 |
| English Summary: |
| To compare the similarity of hepatotoxicity effects of water extract from Genkwa Flos in adult zebrafish and SD rats，and to explore the action mechanism of hepatotoxicity effects of Genkwa Flos.Methods:A total of 320 adult zebrafish were divided into 32 groups(10 in each group，female:male=1:1).High，medium and low dose groups and the blank control group were set up，which were exposed for 96 hours.24 SD rats were divided into 4 groups(6 in each group).The rats of high，medium，low dose group and the blank control group were administered once a day at a dose of 2.5 mL/200 g for 7 consecutive days.The adult zebrafish and rats were then sacrificed at the end of the experiment and liver tissues were collected.The liver morphology was observed through pathological tissue sections.SOD，SDH，MDA，NOX，TG，ALT，AST were used as indicators of toxicity assessment.Based on that，the expressions of apoptosis-related gene in the liver were detected by real-time quantitative PCR.Results:Pathological tissue sections showed that Genkwa Flos could cause deformation，vacuoles，and even apoptosis of the liver cells of zebrafish.At the same time，the water extract from Genkwa Flos could cause loosely arrangement，vacuoles，morphological shrinkage，unclear boundaries，irregular nucleus morphology with hemorrhage of the liver cells of rats.The activity of SOD，SDH and NADH in the liver of adult zebrafish was significantly decreased(P<0.05)，while the activity of MDA was increased(P<0.01).The result of biochemical indexes showed that the activity of SOD，SDH in the liver of rats was significantly decreased(P<0.05)，while the activity of MDA，TG was increased(P<0.01).In addition，the activity of ALT was significantly increased(P<0.05)in low dose group，and the activity of AST was increased(P<0.01)in all drug-administered group.The result of qRT-PCR showed that the expression of genes involved in mitochondrial function(mtatp6，mtco1，mtnd1，pklr)was decreased(P<0.01)in the liver of zebrafish，the mRNA expression of genes involved in oxidative stress(gstp2，nqo1，sod1)was decreased(P<0.01)，the mRNA expression of genes involved in endoplasmic reticulum stress(atf4a，ern1，hsp90b1)was increased(P<0.01)，the mRNA expression of genes involved in autophagy(beclin-1，ATG3)was decreased(P<0.01).Conclusion:The water extract of Genkwa Flos has a toxic effect on the liver of zebrafish adults and SD rats.The mechanism may be that Genkwa Flos can induce oxidative stress damage，mitochondrial dysfunction，endoplasmic reticulum stress and zebrafish liver tissue.Cell autophagy，in turn induces liver cell apoptosis. |
| 查看全文 查看/发表评论 下载PDF阅读器 |
