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1.广西中医药大学,南宁 530200
2.广西中药药效研究重点实验室,南宁 530200
3.广西仙茱中药科技有限公司,南宁 530032
夏玉苹,在读硕士,从事中药质量控制与中药资源开发的研究,E-mail:1114086161@qq.com
* 汝梅,博士,讲师,从事药用植物次生代谢的研究,Tel:0771-4953513,E-mail:rumei2015@163.com; *
李永华,博士,研究员,从事中药质量控制与中药资源开发的研究Tel:0771-4953513,E-mail:liyonghua185@126.com
收稿日期:2021-07-19,
网络出版日期:2021-09-08,
纸质出版日期:2021-11-05
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夏玉苹,何春花,柴子舒等.不同寄主桑寄生水提物对斑马鱼模型的急性毒性及肝损伤[J].中国实验方剂学杂志,2021,27(21):91-97.
XIA Yu-ping,HE Chun-hua,CHAI Zi-shu,et al.Acute Toxicity and Hepatotoxicity of Aqueous Extracts of Taxilli Herba from Different Hosts in Zebrafish Model[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(21):91-97.
夏玉苹,何春花,柴子舒等.不同寄主桑寄生水提物对斑马鱼模型的急性毒性及肝损伤[J].中国实验方剂学杂志,2021,27(21):91-97. DOI: 10.13422/j.cnki.syfjx.20212125.
XIA Yu-ping,HE Chun-hua,CHAI Zi-shu,et al.Acute Toxicity and Hepatotoxicity of Aqueous Extracts of Taxilli Herba from Different Hosts in Zebrafish Model[J].Chinese Journal of Experimental Traditional Medical Formulae,2021,27(21):91-97. DOI: 10.13422/j.cnki.syfjx.20212125.
目的
2
用斑马鱼模型对桑树、漆树、油茶、柳树、苦楝和夹竹桃6种寄主桑寄生水提物进行急性毒性及肝损伤研究,探索寄主对桑寄生毒性的影响,为桑寄生用药安全提供理论依据。
方法
2
以受精后3 d(3 dpf)发育正常的AB系斑马鱼为对象进行急性毒性研究,根据毒性预实验结果,对不同寄主桑寄生水提物分别设置6个不同剂量浓度对斑马鱼进行处理,统计其72 h死亡率,采用GraphPad Prism 6.0软件绘制量-毒曲线,计算不同寄主桑寄生水提物的半数致死浓度(LC
50
)和10%致死浓度(LC
10
)。以受精后4 d(4 dpf)发育正常的gz15Tg/+(AB)肝脏荧光蛋白转基因斑马鱼为对象进行肝损伤研究,将不同寄主桑寄生水提物分别设置低、中、高3个剂量浓度组,设对乙酰氨基酚组,以胚胎水为空白组,给药72 h后观察斑马鱼肝脏形态和荧光面积变化,检测丙氨酸氨基转移酶(ALT)和天冬氨酸氨基转移酶(AST)活性。
结果
2
急性毒性实验结果显示,桑树、漆树、油茶、柳树、苦楝和夹竹桃寄主桑寄生水提物的LC
50
分别为1.24,0.94,0.51,0.38,0.11,0.09 g·L
-1
;LC
10
分别为0.70,0.60,0.35,0.28,0.08,0.07 g·L
-1
。肝损伤实验结果,与空白组比较,对乙酰氨基酚组的斑马鱼的肝脏形态变形和荧光面积减小(
P
<
0.01),ALT和AST活性升高(
P
<
0.01),结果表明对乙酰氨基酚对斑马鱼有肝毒。桑树寄主桑寄生水提物低、中、高3个剂量浓度组斑马鱼的肝脏形态和荧光面积均无变化,ALT和AST活性均降低;漆树、油茶、柳树、苦楝和夹竹桃5种寄主桑寄生水提物中、高剂量浓度组斑马鱼均不同程度地表现出肝脏形态变化和荧光面积减小(
P
<
0.05,
P
<
0.01),ALT和AST活性均明显升高(
P
<
0.05,
P
<
0.01)。结果表明桑树寄主桑寄生对斑马鱼没有肝脏毒性,漆树、油茶、柳树、苦楝和夹竹桃5种寄主桑寄生对斑马鱼表现出不同程度的肝损伤。
结论
2
桑寄生毒性有无或强弱具有寄主依赖性,桑树寄主桑寄生无肝损伤,其他5种寄主桑寄生均不同程度地表现出一定的肝损伤,规范寄主来源可能是实现桑寄生用药安全的重要举措。
Objective
2
To explore the acute toxicities and hepatotoxicities of aqueous extracts of Taxilli Herba from
Morus alba
,
Toxicodendron
trichocarpum
,
Camellia oleifera
,
Salix babylonica
,
Melia azedarach
, and
Nerium indicum
against zebrafish model and the effect of different hosts on the toxicity of Taxilli Herba, hoping to provide a theoretical basis for the safe use of Taxilli Herba.
Method
2
The normally developed AB zebrafish at 3-day post fertilization was selected for acute toxicity study. According to the results of preliminary toxicity experiments, the zebrafishes were treated with aqueous extracts of Taxilli Herba from different hosts at six doses, and their mortality was calculated 72 h later. GraphPad Prism 6.0 was used for plotting the dose-toxicity curve, followed by the calculation of their median lethal concentration (LC
50
) and 10% lethal concentration (LC
10
). The gz15Tg/+(AB) liver fluorescent protein transgenic zebrafish with normal development at 4-day post fertilization was applied for the hepatotoxicity study. The zebrafishes were divided into the low-, medium-, and high-dose groups of aqueous extracts of Taxilli Herba from six hosts, the positive control (acetaminophen) group, and the blank (embryo amniotic fluid) group, and then treated with the corresponding drugs. Seventy-two hours later, the liver morphology and fluorescent area changes in zebrafish were observed. And the activities of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) were detected.
Result
2
The results of acute toxicity test demonstrated that the LC
50
values of water extracts of Taxilli Herba from
M. alba
,
T.
trichocarpum
,
C. oleifera
,
S. babylonica
,
M. azedarach
, and
N. indicum
were 1.24, 0.94, 0.51, 0.38, 0.11, 0.09 g·L
-1
, respectively, and the LC
10
values were 0.70, 0.60, 0.35, 0.28, 0.08, 0.07 g·L
-1
, respectively. As revealed by hepatotoxicity test, compared with the blank group, the positive control group exhibited liver morphological changes, decreased fluorescent area (
P
<
0.01), and elevated ALT and AST activities (
P
<
0.01), suggesting that acetaminophen was hepatotoxic to zebrafish. However, there was no change in the liver morphology or fluorescent area of zebrafish in the low-, medium-, and high-dose groups of water extracts of Taxilli Herba from
M. alba
, and the ALT and AST activities were decreased. By contrast, the liver morphology and fluorescent areas in the medium- and high-dose groups of water extracts of Taxilli Herba from
T.
trichocarpum
,
C. oleifera
,
S. babylonica
,
M. azedarach
, and
N. indicum
changed to varying degrees (
P
<
0.05,
P
<
0.01). Besides, the activities of both ALT and AST were also enhanced. These indicated that Taxilli Herba from
M. alba
had no hepatotoxicity to zebrafish, while that from
T.
trichocarpum
,
C. oleifera
,
S. babylonica
,
M. azedarach
, and
N. indicum
showed varying degrees of hepatotoxicity to zebrafish.
Conclusion
2
The toxicity of Taxilli Herba is host-dependent. Taxilli Herba from
M. alba
has no hepatotoxicity, but that from the other five hosts shows varying degrees of hepatotoxicity. Standardizing the host source may be an important measure to realize the medication safety of Taxilli Herba.
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