白黎芦醇体外对多房棘球蚴原头节和微囊的作用研究  

Effect of resveratrol against Echinococcus multilocularis protoscoleces and microcysts in vitro

作　　者：高海军 庞华胜 张妍 霍乐乐 姜斌 莫筱瑾 雒艳萍[2] 张颋 景涛[2] 徐斌 马兴铭[2] 胡薇 GAO Hai-jun;PANG Hua-sheng;ZHANG Yan;Huo Le-le;JIANG Bin;MO Xiao-jin;LUO Yan-ping;ZHANG Ting;JING Tao;XU Bin;MA Xing-ming;HU Wei(Tibet Autonomous Region Center for Disease Control and Prevention,National Health Commission Key Laboratory of Echinococcosis Prevention and Control,Lhasa 850000,China;School of Basic Medical Sciences,Lanzhou University,Lanzhou 730000,China;National Institute of Parasitic Diseases,Chinese Center for Disease Control and Prevention(Chinese Center for Tropical Diseases Research),NHC Key Laboratory of Parasite and Vector Biology,WHO Collaborating Center for Tropical Diseases,National Center for International Research on Tropical Diseases,Shanghai 200025,China;Garzr Tibetan Autonomous Prefecture Center for Disease Control and Prevention,Kangding 626000,China)

机构地区：[1]西藏自治区疾病预防控制中心,国家卫生健康委员会包虫病防治研究重点实验室,拉萨850000 [2]兰州大学基础医学院,兰州730000 [3]中国疾病预防控制中心寄生虫病预防控制所(国家热带病研究中心)、国家卫生健康委员会寄生虫病原与媒介生物学重点实验室,世界卫生组织热带病合作中心,国家级热带病国际联合研究中心,上海200025 [4]甘孜藏族自治州疾病预防控制中心,康定626000

出　　处：《中国人兽共患病学报》2021年第12期1057-1063,共7页Chinese Journal of Zoonoses

基　　金：中国医学科学院中央级公益性科研院所基本科研业务费专项资金(No.2019PT320004);国家自然科学基金(No.31860697、No.81171632、No.81201315);上海市卫生健康委员会面上项目(No.201840133)。

摘　　要：目的研究白藜芦醇(Resveratrol,RES)体外对多房棘球蚴原头节(Protoscoleces,PSCs)和微囊的作用效果,为多房棘球蚴病的临床用药提供新的依据。方法PSCs体外经不同浓度(5、10、20、40、80、100、200和400μmol/L)的RES作用7 d,采用台盼蓝染色观察其活力与形态变化,采用扫描电镜和透射电镜观察其超微结构变化,运用化学发光法检测上清中碱性磷酸酶(Alkaline phosphatase,ALP)活性;微囊体外经RES(40μmol/L)作用14 d后于光镜下观察其活力和形态变化,运用化学发光法检测上清中ALP活性。结果不同浓度的RES体外持续作用7 d,PSCs死亡率呈剂量、时间依赖性增高,并伴随大量小钩脱落。当40μmol/L RES作用至第6 d,PSCs死亡率达到100%,且虫体小钩、吸盘和外皮等超微结构被破坏;在同条件下,阿苯达唑亚砜(Albendazole sulfoxide,ABZ-SO)处理组中PSCs死亡率为13.25%±1.41%,对照组-二甲基亚砜(Dimethyl sulfoxide,DMSO)组PSCs死亡率仅为6%±0.71%。不同浓度的RES作用至第6 d,PSCs培养上清中ALP活性呈剂量依赖性升高(F=36.94,P<0.001),其中,RES浓度为20和40μmol/L时,上清中ALP活性分别为(2.72±0.24)U/L和(2.95±0.10)U/L,分别与DMSO组(1.97±0.18)U/L间差异存在统计学意义(t=2.94,P=0.0259和t=7.066,P=0.0004)。另外,RES作用至14 d,微囊透光性降低,生发层皱缩且与角质层分离,同时上清中ALP活性(2.74±0.15)U/L高于较DMSO组(2.08±0.09)U/L(t=3.83,P=0.019),但ABZ-SO组(2.29±0.14)U/L与DMSO组间差异无统计学意义(t=1.271,P=0.273)。结论白藜芦醇体外可抑制多房棘球蚴原头节和微囊的活性,有望成为治疗多房棘球蚴病的潜在候选药物。To investigate the effect of resveratrol(RES)against protoscoleces(PSCs)and microcysts of Echinococcus multilocularis in vitro,providing the new references for drugs application to treat alveolar echinococcosis.The vitality and morphological changes in PSCs exposed to RES at different concentrations(5,10,20,40,80,100,200 or 400μmol/L)for 7 days in vitro were investigated by staining with 0.4%trypan blue,and the ultrastructural alterations were imaged by scanning electron microscopy(SEM)and transmission electron microscopy(TEM),respectively;alkaline phosphatase(ALP)activity in culture supernatants was measured using chemiluminescence method.Further,the viability and morphology of E.multilocularis microcysts after treatment with RES for 14 days in vitro were investigated under a light microscopy,and then ALP activity in culture supernatant was measured using chemiluminescence method.The mortality of E.multilocularis PSCs increased with a dose-and time-dependent manner after treatment with RES at 5 to 400μmol/L during 7 days in vitro,accompanying with obvious hooks loss.The mortality of PSCs reached 100%after treatment with 40μmol/L RES for 6 days,and the ultrastructure of suckers,hooks and tegument were destroyed,however,under the same conditions,the mortality of PSCs was only(13.25±1.41)%and(6±0.71)%after treatment with ABZ-SO and dimethye sulfoxide(DMSO),respectively.The ALP activity in culture supernatant of PSCs significantly increased with a dose-dependent manner(F=36.94,P<0.001)on 6^(th) day at different concentrations of RES.When treated with 20 or 40μmol/L RES,the ALP activity was(2.72±0.24)U/L and(2.95±0.10)U/L,respectively,which posed significant differences in comparison with DMSO treatment(1.97±0.18)U/L(t=2.94,P=0.026 and t=7.066,P=0.000).In addition,after treatment with 40μmol/L RES for 14 days,light transmittance in E.multilocularis microcysts deceased observably,and the germinal layer was shrunk and separated from laminated layer.Meanwhile,ALP activity(2.74±0.15)U/L in RES group was observ

关 键 词：白黎芦醇 多房棘球蚴 原头节 微囊 碱性磷酸酶 

分 类 号：R383.3[医药卫生—医学寄生虫学]