中国寄生虫学与寄生虫病杂志 ›› 2010, Vol. 28 ›› Issue (4): 20-320.

• 研究简报 • 上一篇    下一篇

刚地弓形虫速殖子与鼠星形胶质细胞体外共培养的实验观察

李冬娜1,梁幼生2,周永华2,张焕相1,沈海英1,骆伟1,龚唯1,诸葛洪祥1 *   

  1. 苏州大学独墅湖校区病原生物学系教研室,苏州 215123
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-08-30 发布日期:2010-08-30

In vitro Co-cultivation of Toxoplasma gondii Tachyzoites with Rat Brain Astrocytes

LI Dong-na1,LIANG You-sheng2,ZHOU Yong-hua2,ZHANG Huan-xiang1,
SHENG Hai-ying1,LUO Wei1,GONG Wei1,ZHUGE Hong-xiang 1 *
  

  1. 1 Department of Pathobiology,Soochow University,Suzhou 215123,China;2 Jiangsu Institute of Parasitic Disease,Wuxi 214064,China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-08-30 Published:2010-08-30

摘要: 原代培养星形胶质细胞,待其长至培养皿的80%时,将星形胶质细胞与弓形虫速殖子(细胞与虫体的比例为1 ∶ 10)共培养0~72 h,吉氏染色观察星形胶质细胞和弓形虫速殖子的变化。共培养1 h,此时定为0点,再培养0~48 h,单磺酰戊二胺染色在荧光显微镜下观察星形胶质细胞内自噬囊泡的变化。结果发现共培养1 h时,弓形虫速殖子开始进入星形胶质细胞,并出现荧光颗粒;8 h星形胶质细胞内荧光颗粒最多;12 h后显著减少,星形胶质细胞开始被破坏;48 h星形胶质细胞内荧光颗粒消失;72 h大部分星形胶质细胞已被涨破,留下大量增殖的弓形虫速殖子。说明自噬在弓形虫速殖子体外感染星形胶质细胞的过程中是受到抑制的。

关键词: 星形胶质细胞, 弓形虫, 速殖子, 自噬

Abstract: Purified astrocytes were cultured in plates. When astrocytes grew over 80% of the plate, tachyzoites of Toxoplasma gondii RH strain were added for co-culture. In the period of 0-72 h, change of the astrocytes and tachyzoites was observed after Giemsa staining. In 0-48 h, monodansylcadaverine (MDC) was used to study the action of autophagy in the process of tachyzoites invading astrocytes. At 1 h co-culture, tachyzoites had entered in astrocytes and the autophagosomes appeared. At 4 h, the autophagosomes increased pronouncedly. However, after 12 h, number of autophagosomes considerably decreased and damage of the cells occurred. 48 h later, autophagosomes disappeared and more astrocytes were destroyed. At 72 h most cells destroyed and tachyzoites were released. The result showed that autophagy is inhibited when the astrocytes were in vitro infected by tachyzoites.

Key words: Astrocyte, Toxoplasma gondii, Tachyzoite, Autophagy