Inhibition of botulinum toxin's association with rat brain synaptosomes by toosendanin,an antibotulismic triterpenoid  

作　　者：Jianying Zhou Yuliang Shi 

机构地区：[1]Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, Shanghai 200031, China

出　　处：《Neural Regeneration Research》2006年第6期489-492,共4页中国神经再生研究（英文版）

基　　金：the Major State Basic Research Development Program of China (973 pro-gram), No. G1999054000; the National Natural Science Foun-dation of China, No. 30170302;Basic Research Program of Shanghai, No. 02JC14011

摘　　要：BACKGROUND： Toosendanin （TSN）, a tnterpenoid derivative extracted from the bark of Melia toosendanin Sieb et Zucc, has been demonstrated to be an effective drug for treatment of experimental botulism. OBJECTIVE ： To explore its antibotulismic mechanism by observing the effect of toosendanin on association of botulinum toxin （BoTx） with rat brain synaptosomes under different conditions. DESIGN： A randomized controlled experiment SETTING： Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences MATERIALS： Sprague-Dawiey rats, weighing （220±12） g, were involved. TSN was a sample recrystallized in ethanol with a punty 〉 98%. BoTx/A and BoTx/C of 500 ku （1.8×10^7 mouse LD50/mg） were purchased from Wako （Japan）, Horse antitoxins to BoTx/A and/C （primary antibodies） were purchased from Lanzhou Institute of Biological Products （China）. METHODS ： Major experiments were finished between March 2005 and October 2005 in key laboratory of neurobiology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. （1） Preparation of synaptosome of rats： one aliquot of synaptosome was pre-incubated with TSN （17 μmol/L） for 20 minutes at 4 % or 37 % （another aliquot of synaptosome which was untouched was used as control）. Then, BoTx was respectively added for another 20-minute incubation. （2） A homochronous expenment was still performed at 37 %. Differently, high level of K^＋ was used to stimulate synaptosome for 25 minutes. Two aliquots of synaptosomes with or without TSN （17 μmol/L）were preincubated for 15 minutes, Then, 30 mmol/L KCl was separately added in two aliquots, 5 minutes later, 13 nmol/L BoTx/C was separately added followed by 20-minute incubation. （3）The effect of TSN on BoTx binding was observed by Western blot and synchronization method. （4） t test was used for comparing the difference of measurement data. MAIN OUTCOME MEASURES ： The gray value of BoTx bands from western blot was used to estimate the bound aBACKGROUND： Toosendanin （TSN）, a tnterpenoid derivative extracted from the bark of Melia toosendanin Sieb et Zucc, has been demonstrated to be an effective drug for treatment of experimental botulism. OBJECTIVE ： To explore its antibotulismic mechanism by observing the effect of toosendanin on association of botulinum toxin （BoTx） with rat brain synaptosomes under different conditions. DESIGN： A randomized controlled experiment SETTING： Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences MATERIALS： Sprague-Dawiey rats, weighing （220±12） g, were involved. TSN was a sample recrystallized in ethanol with a punty 〉 98%. BoTx/A and BoTx/C of 500 ku （1.8×10^7 mouse LD50/mg） were purchased from Wako （Japan）, Horse antitoxins to BoTx/A and/C （primary antibodies） were purchased from Lanzhou Institute of Biological Products （China）. METHODS ： Major experiments were finished between March 2005 and October 2005 in key laboratory of neurobiology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences. （1） Preparation of synaptosome of rats： one aliquot of synaptosome was pre-incubated with TSN （17 μmol/L） for 20 minutes at 4 % or 37 % （another aliquot of synaptosome which was untouched was used as control）. Then, BoTx was respectively added for another 20-minute incubation. （2） A homochronous expenment was still performed at 37 %. Differently, high level of K^＋ was used to stimulate synaptosome for 25 minutes. Two aliquots of synaptosomes with or without TSN （17 μmol/L）were preincubated for 15 minutes, Then, 30 mmol/L KCl was separately added in two aliquots, 5 minutes later, 13 nmol/L BoTx/C was separately added followed by 20-minute incubation. （3）The effect of TSN on BoTx binding was observed by Western blot and synchronization method. （4） t test was used for comparing the difference of measurement data. MAIN OUTCOME MEASURES ： The gray value of BoTx bands from western blot was used to estimate the bound a

分 类 号：R74[医药卫生—神经病学与精神病学]