外源宿主培养模式对细粒棘球蚴原头节活性的影响

Effects of exogenous host culture model on the activity of the Echinococcus granulosus protoscoleces in vitro

目的明确外源宿主在何种体外培养模式下对细粒棘球蚴原头节活性具有维持作用。方法收集HepG-2肝癌细胞及其无血清上清液,与细粒棘球蚴原头节(每组约7000个)建立共培养体系,分别为:对照组、HepG-2细胞组、HepG-2上清组和HepG-2细胞+上清组。在第1、4、7、11d,取各组混悬液70μl台盼蓝染色计算存活率。ELISA法检测各组原头节半胱氨酸蛋白酶-3酶的活力单位。结果各组原头节的存活率随时间逐渐降低,第11d HepG-2细胞组、HepG-2细胞+上清组和HepG-2上清组较对照组均存在统计学意义(PHepG-2细胞组、HepG-2细胞+上清组、HepG-2上清组<0.01)。第11d,HepG-2上清组和HepG-2细胞+上清组相对于对照组在半胱氨酸蛋白酶-3酶活性上差异均有统计学意义(PHepG-2上清、HepG-2细胞+上清<0.01)。结论外源宿主体外培养下对细粒棘球蚴原头节活性的维持以宿主细胞上清形式最显著。

Objective To identify the maintenance effect of the exogenous host in vitro culture on the activity of the Echinococcus granulosus protoscoleces.Methods HepG-2 cells and their serum-free supernatant were collected and co-cultured with echinococcus granulosus(about 7,000 in each group).All cells and supernatant were divided into the control group,HepG-2 cell group,HepG-2 supernatant group and HepG-2 cell plus supernatant group,respectively.On day 1,4,7 and 11 th,70μl trypan blue was used to calculate the survival rate of each group.The activity units of Caspase-3 in each group were determined by ELISA method.Results The survival rate of the protoscoloces of each group gradually decreased with time.On the 11 th day,the HepG-2 cell group,HepG-2 cell plus supernatant group and HepG-2 supernatant group all had statistical significance(PHepG-2 cell Croup,HepG-2 cells+supernatant group,HepG-2 supematant group<0.01).On the 11 th day,there were statistically significant differences in Caspase-3 enzyme activity between the HepG-2 supernatant group and the HepG-2 cell+supernatant group compared to the control group(PHepG-2 supernatant,HepG-2 cell+supmatant<0.01).Conclusions The supernatant of exogenous host cells was the most significant factor to maintain the activity of Echinococcus granulosus protoscoleces in vitro culture.