摘要
目的探讨泡球蚴囊液对体外原代培养大鼠肝脏细胞增殖的影响。方法以Wistar大鼠作肝细胞供者,采用胶原酶肝脏原位灌流消化法分离肝细胞。取1×106肝细胞接种于Ⅰ型胶原铺底35mm培养皿,无血清培养20h后以泡球蚴囊液置换培养液培养24h和48h,期间在荧光倒置显微镜下观察肝细胞形态变化,流式细胞仪测定细胞周期;另取1×105肝细胞接种于Ⅰ型胶原铺底96孔培养板,同上培养,MTT法检测泡球蚴囊液对大鼠肝脏细胞毒性作用。结果与泡球蚴囊液共培养后肝细胞贴壁生长良好,MTT法测定肝细胞存活率为77.5%～100%。培养24h后肝细胞增殖指数(PI)泡球蚴囊液处理组为(49.35±4.00)%,对照组为(82.00±6.00)%;培养48h后两组肝细胞增殖指数(PI)分别为(27.61±6.00)%和(67.00±10.00)%,差异均有显著性(P<0.05)。结论泡球蚴囊液对体外培养大鼠肝细胞无毒害作用,但对肝细胞的增殖有一定的抑制作用。
Objective To study the effect of cyst fluid of the parasite Echinococcus multilocularis on proliferation of rat hepatocytes in vitro. Methods Hepatocytes were isolated from Wistar rat by collagenase digestion and 1×10^6 cells were cultured in 35 mm culture capsule coated with collagen I. After cultured in a conditioned medium without serum for 20 h, hepatocytes were incubated with cyst fluid for 24 h and 48 h, respectively. The morphologic change of cultured hepatocytes was observed by fluorescence microscope, the cell cycle was assessed by flow cytometry. Hepatocytes,cultured in 96 culture plate＇coated with collagen I at 1×10^5 , the same culture method, toxic effect on hepatocytes by cyst fluid was assayed with MTT method. Results Hepatocytes incubated with cyst fluid were intact and have a good function. The MTT assay showed the livability of hepatocytes was from 77.5% to 100%. The proliferation index （PI） of hepatocytes cultured with cyst fluid was （49.35±4.00） %, and the PI of the control group was （82.00±6.00） % after 24 h, and the PI of hepatocytes was （27.61 ± 6.00）% and （67.00 ± 10.00）% after 48 h. The two group were different significantly （P 〈0.05）. Conclusion The cyst fluid had no inhibition effect on primary cultured rat hepatocytes, but it could inhibit the proliferation of hepatocytes.
出处
《中国病原生物学杂志》
CSCD
2008年第3期197-199,F0004,共4页
Journal of Pathogen Biology
基金
新疆维吾尔自治区高校科研创新研究群体基金资助项目(NoXJEDU2004G10)
新疆重点实验室开放课题基金资助项目(NoXJDX0202-2005-01)
关键词
泡球蚴囊液
大鼠肝细胞
MTT
细胞增殖
Echinococcus multilocularis cyst fluid
rat hepatocytes
MTT
cell proliferation
