右旋松萝酸体外抗弓形虫速殖子的实验研究  

In vitro Effect of(+)-Usnic Acid on Toxoplasm gondii Tachyzoites

作　　者：魏琳琳[1] 程彦斌[1] 司开卫[1] 顾宁[1] 李小其[1] 李琛[1] 袁育康[1] 

机构地区：[1]西安交通大学医学院免疫学与病原生物学系,西安710061

出　　处：《中国寄生虫学与寄生虫病杂志》2008年第6期438-441,共4页Chinese Journal of Parasitology and Parasitic Diseases

基　　金：国家自然科学基金(No.30571628);陕西省自然科学基金(No.2004C228);陕西省科技攻关基金(No.2004K10-G6-2)~~

摘　　要：目的探索右旋松萝酸在体外抗RH株弓形虫速殖子的效果。方法实验设右旋松萝酸组、乙酰螺旋霉素组、二甲基亚砜(DMSO)组及生理盐水对照组,前两组分别再设4个浓度组,于1ml弓形虫速殖子悬液(1×106个/ml)中分别加入终浓度为5、10、25和50μg/ml右旋松萝酸或乙酰螺旋霉素;后两组分别加入等体积的生理盐水和1%DMSO;每组15管。作用1、2和4h后,取弓形虫悬液涂片,台盼蓝染色,光镜观察记数。将各组作用4h的弓形虫悬液洗涤后,接种昆明小鼠,作再转种试验,观察弓形虫速殖子复苏情况。同时将洗涤后的虫体接种于制备的SD大鼠心肌成纤维细胞,观察虫体侵袭力。结果10、25和50μg/ml右旋松萝酸组作用4h,弓形虫速殖子台盼蓝着色率达100%,部分虫体肿胀,两端变圆,细胞质内出现颗粒,细胞核深染。各浓度的乙酰螺旋霉素组弓形虫速殖子变化与右旋松萝酸组相似,仅50μg/ml组作用4h后台盼蓝着色率达100%。再转种试验结果显示右旋松萝酸组仅5μg/ml组小鼠于接种后第8～9天死亡,腹腔液见大量弓形虫速殖子;其余各组小鼠大多存活至再次转种,且腹腔液均未见弓形虫速殖子。但乙酰螺旋霉素各浓度组小鼠均于接种后6～8d死亡,腹腔液见大量虫体及假包囊。右旋松萝酸5μg/ml组,弓形虫速殖子侵入大鼠心肌成纤维样细胞,会形成假包囊,其余组均为阴性。但乙酰螺旋霉素、生理盐水和DMSO组速殖子对大鼠心肌成纤维细胞均有不同程度感染。结论右旋松萝酸在体外有较强的抗弓形虫速殖子作用。Objective To explore the effect of usnic acid on Toxoplasma gondii tachyzoites in vitro. Methods There are four groups named as （＋）-usnic acid group, acetylespiramycin group, DMSO group and normal saline group. Groups of （＋）-usnic acid and acetylespiramycin were further divided into 4 subgroups with final concentration of 5, 10, 25, 50 μg/ml respectively. Normal saline group and DMSO group were respectively given equal volume normal saline and 1% DMSO. Each group have 15 parallel tubes with 1 ml （1×10^6/ml） T. gondii tachyzoites aqueous suspension. At 1 h, 2h and 4 h after drug treatment, tachyzoites were counted by light microscope with 0.4% Trypan blue staining. Tachyzoites in aqueous suspension was collected, and washed 3 times by PBS solution. Normal mice were inoculated intraperitoneally and observed for three generations. The cultivated rat cardiofibroblasts were then infected in vitro with T. gondii tachyzoites. At the same time, rat cardiomycytes invasion by T. gondii tachyzoites was investigated. Results At 4 h treated by 10, 25 and 50 μg/ml （＋）-usnic acid, 100% T. gondii tachyzoites were stained. Some tachyzoites were swelling, blunt or round in the two ends; and granules appeared in the cytoplasm, the nuclei were deep stained. The changes of tachyzoites in acetylespiramycin group were similar to （＋）-usnic acid group, 100% T. gondii taehyzoites were stained in 50 μg/ml acetylespiramyein subgroup. In inoculation tests, mice died at 8th to 9th days in 5μg/ml （＋）-usnic acid subgroup and numerous taehyzoites were detected in ascites. However, most mice survived to be killed in the other （＋）-usnie acid subgroups and the tachyzoites were not found in aseites. All mice in acetylespirmycin groups died at 6th to 8th days after inoculation and many tachyzoites or psudocysts were observed in mice ascites. In infecting cell tests, the cultivated rat cardiofibroblasts were infected in vitro by the tachyzoites after treated with 5μg/ml （＋）-usnic acid for 4 h, and ps

关 键 词：松萝酸 乙酰螺旋霉素 弓形虫 速殖子 

分 类 号：R382.5[医药卫生—医学寄生虫学]