摘要
为探究苍术、川乌等12种中草药的体外抗弓形虫感染效果,经制备水煎剂、测取吸光值和干重,评价其对Vero细胞的毒害作用;合成引物,优化real-time PCR条件,用9种回归模型拟合不同感染孔内弓形虫数目,求得24孔板单孔适合感染量;用适合数目RH速殖子与适当浓度水煎剂作用6 h、12 h、1 d、2 d和3 d,real-time PCR法定量水煎剂的体外抗弓形虫效果。结果显示,所得水煎剂的吸光值大小与干重间无线性关系,高浓度水煎剂不利于Vero细胞生长。抗感染结果显示,槟榔、白薇、青蒿、蒲公英和柴胡作用3 d时弓形虫速殖子数均较空白组显著降低（P〈0.05）,其他水煎剂和时间点则作用不明显（P〉0.05）,表明槟榔、白薇、青蒿、蒲公英和柴胡可抑制弓形虫体外增殖,能用作研发抗弓形虫感染特效药的重要研究对象。
To evaluate in vitro anti-Toxoplasma gondii efficacy of twelve kinds of Chinese herbal medicines,their optical densities,dry weights and toxicities for Vero cells were examined by preparing the boil preparations.PCR amplifications were performed to identify the synthesized primers and the real-time PCR conditions.Based on nine kinds of frequent regression models,the curves were matched according to the number of T.gondii parasites in wells,and their parameters were further used to calculate the suitable infection doses of this parasite for 24-well plate.Anti-T.gondii efficacy of boilpreparations was evaluated by the created real-time PCR at 6 h,12 h,1 d,2 d, and 3 d post infection.The data showed that the same dry weight of boil preparations own significantly different optical densities,and the host cell growth was severely affect by higher doses.The results of real-time PCR indicated that the numbers of T.gondii were statistically reduced at 3 days post preparation addition in Semen Arecae catechu-,Radix Cynanchi atrati-,Herba Artemisiae apiaceae-,Hr Teraxaci mongolicicum radiceor Radix Bupleuri-contained wells compared with the controls（P〈0.05）,and no difference was detected at other time points and the last seven kinds of boil preparations（P〈0.05）,suggesting that in vitro proliferation of T.gondii was significantly inhibited by Semen Arecae catechu,Radix Cynanchi atrati,Herba Artemisiae apiaceae,Hr Teraxaci mongolicicum radice and Radix Bupleuri,which could be further used to exploit the miracle drugs for the clinical treatments of toxoplasmosis.
作者
郭海婷
魏艳梅
陈新亮
朱兴全
李中原
陈志宝
GUO Hai-ting;WEI Yan-mei;CHEN Xin-liang;ZHU Xing-quan;LI Zhong-yuan;CHEN Zhi-bao(College of Biological Science and Technology,Heilong~iang Bayi Agricultural University,Doqing 163319,China;State Key Laboratory of Veterinary Etiological Biology~Key Laboratory of Veterinary Parasitology of Gansu Province/ Lanzhou Veterinary Research Institute,Chinese Academy of Agricultural Sciences,Lanzhou 730046,China;College of Animal Science and Technology,A nhui Agricultural University,Hefei 230036,China)
出处
《中国兽医科学》
CAS
CSCD
北大核心
2018年第12期1569-1576,共8页
Chinese Veterinary Science
基金
安徽省博士后研究人员科研活动经费资助项目(2017B188)
中国博士后科学基金面上资助项目(2017M612056)
家畜疫病病原生物学国家重点实验室开放基金课题(SKLVEB2016KFKT015)
