【摘要】：目的應用模式生物秀麗隱桿線蟲研究中藥芫花的主要成分之一芫花素的毒性作用及機制。方法分別以芫花素0.1,1,10和100μmol·L~(-1)處理野生型或轉(zhuǎn)基因型秀麗隱桿線蟲(簡稱線蟲)24,48和72 h,顯微鏡下觀察野生型線蟲頭部擺動次數(shù)、身體彎曲次數(shù)、存活天數(shù)、后代數(shù)目、平均排泄循環(huán)長度和咽泵行為,并觀察轉(zhuǎn)基因型線蟲γ-氨基丁酸(GABA)能神經(jīng)元熒光強度,計數(shù)神經(jīng)元數(shù)目。結(jié)果 (1)野生型線蟲:與正常對照組比較,芫花素處理24 h后,芫花素1~100μmol·L~(-1)組線蟲頭部擺動和身體彎曲次數(shù)顯著減少(P0.05,P0.01);48 h后,芫花素0.1μmol·L~(-1)組身體彎曲次數(shù)顯著減少(P0.05),1~100μmol·L~(-1)組頭部擺動和身體彎曲次數(shù)顯著減少(P0.01),100μmol·L~(-1)組平均排泄循環(huán)長度顯著增加(P0.05)且咽泵行為顯著減少(P0.01);72 h后,芫花素0.1~100μmol·L~(-1)組頭部擺動和身體彎曲次數(shù)均顯著減少(P0.05,P0.01),平均排泄循環(huán)長度顯著增加(P0.05,P0.01),咽泵行為顯著減少(P0.05,P0.01)。(2)轉(zhuǎn)基因型線蟲:與正常對照組比較,芫花素處理24 h后,芫花素1~100μmol·L~(-1)組轉(zhuǎn)基因型線蟲GABA能神經(jīng)元熒光強度顯著降低(P0.05,P0.01);48 h后,芫花素1~100μmol·L~(-1)組GABA能神經(jīng)元熒光強度顯著降低(P0.05,P0.01)且VD神經(jīng)元數(shù)目顯著減少(P0.01);72 h后,芫花素0.1~100μmol·L~(-1)組GABA能神經(jīng)元熒光強度顯著降低(P0.05,P0.01),1~100μmol·L~(-1)組AVL神經(jīng)元數(shù)目顯著減少(P0.05,P0.01),10和100μmol·L~(-1)組VD神經(jīng)元數(shù)目顯著減少(P0.05,P0.01)。結(jié)論芫花素對秀麗隱桿線蟲具有一定的毒性作用,主要表現(xiàn)為運動行為抑制,其毒性機制可能與GABA能神經(jīng)元損傷有關。
[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 緯-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.
【作者單位】： 南京中醫(yī)藥大學江蘇省中藥資源產(chǎn)業(yè)化過程協(xié)同創(chuàng)新中心;南京中醫(yī)藥大學藥物安全性評價研究中心;
【基金】：國家自然科學基金(81202981) 江蘇省中藥資源產(chǎn)業(yè)化過程協(xié)同創(chuàng)新中心開放課題(ZDXM-2-7)~~
【分類號】：R114
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本文編號：2447558