芫花素对模式生物秀丽隐杆线虫的毒性作用
发布时间：2019-03-26 12:46
【摘要】：目的应用模式生物秀丽隐杆线虫研究中药芫花的主要成分之一芫花素的毒性作用及机制。方法分别以芫花素0.1,1,10和100μmol·L~(-1)处理野生型或转基因型秀丽隐杆线虫(简称线虫)24,48和72 h,显微镜下观察野生型线虫头部摆动次数、身体弯曲次数、存活天数、后代数目、平均排泄循环长度和咽泵行为,并观察转基因型线虫γ-氨基丁酸(GABA)能神经元荧光强度,计数神经元数目。结果 (1)野生型线虫:与正常对照组比较,芫花素处理24 h后,芫花素1~100μmol·L~(-1)组线虫头部摆动和身体弯曲次数显著减少(P0.05,P0.01);48 h后,芫花素0.1μmol·L~(-1)组身体弯曲次数显著减少(P0.05),1~100μmol·L~(-1)组头部摆动和身体弯曲次数显著减少(P0.01),100μmol·L~(-1)组平均排泄循环长度显著增加(P0.05)且咽泵行为显著减少(P0.01);72 h后,芫花素0.1~100μmol·L~(-1)组头部摆动和身体弯曲次数均显著减少(P0.05,P0.01),平均排泄循环长度显著增加(P0.05,P0.01),咽泵行为显著减少(P0.05,P0.01)。(2)转基因型线虫:与正常对照组比较,芫花素处理24 h后,芫花素1~100μmol·L~(-1)组转基因型线虫GABA能神经元荧光强度显著降低(P0.05,P0.01);48 h后,芫花素1~100μmol·L~(-1)组GABA能神经元荧光强度显著降低(P0.05,P0.01)且VD神经元数目显著减少(P0.01);72 h后,芫花素0.1~100μmol·L~(-1)组GABA能神经元荧光强度显著降低(P0.05,P0.01),1~100μmol·L~(-1)组AVL神经元数目显著减少(P0.05,P0.01),10和100μmol·L~(-1)组VD神经元数目显著减少(P0.05,P0.01)。结论芫花素对秀丽隐杆线虫具有一定的毒性作用,主要表现为运动行为抑制,其毒性机制可能与GABA能神经元损伤有关。
[Abstract]:Objective to study the toxicity and mechanism of Daphne genkwa, one of the main components of traditional Chinese medicine (Daphne genkwa), by using model organism Cryptococcus elegans. Methods Genkwa genkwa 0.1, 1, 10 and 100 渭 mol 路L ~ (- 1) were treated with wild-type or transgenic nematode (nematode) for 24, 48 and 72 hours, respectively. The head swings and body bending times of wild-type nematodes were observed under microscope. The survival days, the number of offspring, the average excretory circulation length and pharyngeal pumping behavior were observed. The fluorescence intensity of 纬-aminobutyric acid (GABA) neurons in transgenic nematode was observed and the number of neurons was counted. Results (1) Wild-type nematode: compared with the normal control group, the number of head swings and body bending of Daphne genkwa treated with 1 ~ 100 渭 mol 路L ~ (- 1) Daphne genkwa decreased significantly after 24 h treatment (P 0.05, P0.01). After 48 hours, the number of body bending in 0.1 渭 mol 路L ~ (- 1) Daphne genkwa group decreased significantly (P0.05), and the number of head swings and body bending decreased significantly in 1 ~ 100 渭 mol 路L ~ (- 1) group (P0.01). In 100 渭 mol 路L ~ (- 1) group, the average excretory circulation length was significantly increased (P0.05) and pharyngeal pump behavior was significantly decreased (P0.01). After 72 hours, the number of head swings and body bending in 0.1 ~ 100 渭 mol 路L ~ (- 1) Daphne genkwa group decreased significantly (P 0.05, P0.01), and the average excretory circulation length increased significantly (P 0.05, P0.01). Significant decrease in pharyngeal pump behavior (P0.05, P0.01). (2): compared with the normal control group, after 24 h treatment with genkwa, there was no significant difference between the control group and the control group (P 0.05, P0.01). In 1 ~ 100 渭 mol 路L ~ (- 1) of Daphne genkwa group, the fluorescence intensity of GABA neurons in transgenic nematode was significantly decreased (P0.05, P0.01). 48 h later, the fluorescence intensity of GABA neurons and the number of VD neurons were significantly decreased in 1 ~ 100 渭 mol 路L ~ (- 1) Daphne genkwa group (P 0.05, P0.01). After 72 hours, the fluorescence intensity of GABA neurons in 0.1 ~ 100 渭 mol 路L ~ (- 1) group was significantly decreased (P 0.05, P0.01), and the number of AVL neurons in 1 ~ 100 渭 mol 路L ~ (- 1) group was significantly decreased (P 0.05, P0.01). The number of VD neurons decreased significantly in 10 and 100 渭 mol 路L ~ (- 1) groups (P 0.05, P0.01). Conclusion Daphne genkwa has some toxic effects on nematode, the main manifestation is motor behavior inhibition, and its toxic mechanism may be related to the damage of GABA-denergic neurons.
【作者单位】： 南京中医药大学江苏省中药资源产业化过程协同创新中心;南京中医药大学药物安全性评价研究中心;
【基金】：国家自然科学基金(81202981) 江苏省中药资源产业化过程协同创新中心开放课题(ZDXM-2-7)~~
【分类号】：R114
，
本文编号：2447558
[Abstract]:Objective to study the toxicity and mechanism of Daphne genkwa, one of the main components of traditional Chinese medicine (Daphne genkwa), by using model organism Cryptococcus elegans. Methods Genkwa genkwa 0.1, 1, 10 and 100 渭 mol 路L ~ (- 1) were treated with wild-type or transgenic nematode (nematode) for 24, 48 and 72 hours, respectively. The head swings and body bending times of wild-type nematodes were observed under microscope. The survival days, the number of offspring, the average excretory circulation length and pharyngeal pumping behavior were observed. The fluorescence intensity of 纬-aminobutyric acid (GABA) neurons in transgenic nematode was observed and the number of neurons was counted. Results (1) Wild-type nematode: compared with the normal control group, the number of head swings and body bending of Daphne genkwa treated with 1 ~ 100 渭 mol 路L ~ (- 1) Daphne genkwa decreased significantly after 24 h treatment (P 0.05, P0.01). After 48 hours, the number of body bending in 0.1 渭 mol 路L ~ (- 1) Daphne genkwa group decreased significantly (P0.05), and the number of head swings and body bending decreased significantly in 1 ~ 100 渭 mol 路L ~ (- 1) group (P0.01). In 100 渭 mol 路L ~ (- 1) group, the average excretory circulation length was significantly increased (P0.05) and pharyngeal pump behavior was significantly decreased (P0.01). After 72 hours, the number of head swings and body bending in 0.1 ~ 100 渭 mol 路L ~ (- 1) Daphne genkwa group decreased significantly (P 0.05, P0.01), and the average excretory circulation length increased significantly (P 0.05, P0.01). Significant decrease in pharyngeal pump behavior (P0.05, P0.01). (2): compared with the normal control group, after 24 h treatment with genkwa, there was no significant difference between the control group and the control group (P 0.05, P0.01). In 1 ~ 100 渭 mol 路L ~ (- 1) of Daphne genkwa group, the fluorescence intensity of GABA neurons in transgenic nematode was significantly decreased (P0.05, P0.01). 48 h later, the fluorescence intensity of GABA neurons and the number of VD neurons were significantly decreased in 1 ~ 100 渭 mol 路L ~ (- 1) Daphne genkwa group (P 0.05, P0.01). After 72 hours, the fluorescence intensity of GABA neurons in 0.1 ~ 100 渭 mol 路L ~ (- 1) group was significantly decreased (P 0.05, P0.01), and the number of AVL neurons in 1 ~ 100 渭 mol 路L ~ (- 1) group was significantly decreased (P 0.05, P0.01). The number of VD neurons decreased significantly in 10 and 100 渭 mol 路L ~ (- 1) groups (P 0.05, P0.01). Conclusion Daphne genkwa has some toxic effects on nematode, the main manifestation is motor behavior inhibition, and its toxic mechanism may be related to the damage of GABA-denergic neurons.
【作者单位】： 南京中医药大学江苏省中药资源产业化过程协同创新中心;南京中医药大学药物安全性评价研究中心;
【基金】：国家自然科学基金(81202981) 江苏省中药资源产业化过程协同创新中心开放课题(ZDXM-2-7)~~
【分类号】：R114
，
本文编号：2447558
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