问号钩端螺旋体诱导的Vero和J774A.1细胞的凋亡和超微结构病变  

Apoptosis and ultrastructural lesions in Vero and J774A.1 cells induced by Leptospira interrogans

作　　者：李立伟[1] 刘云英[1] 严杰[1] 毛亚飞[1] 罗依惠[1] 李淑萍[1] 

机构地区：[1]浙江大学医学院病原生物学教研室,浙江杭州310031

出　　处：《浙江大学学报（医学版）》2005年第1期4-8,共5页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金项目 (30 170 0 4 9)

摘　　要：目的 :探讨不同毒力的钩体黏附和侵入宿主细胞能力及其病理变化。方法 :建立 Fontana镀银染色法用于观察问号钩体黄疸出血群赖型 5 6 6 0 1株、波摩那群波摩那型 5 6 6 0 8株和双曲钩体三堡隆群 patoc型 Patoc 株黏附 Vero和 J774 A.1细胞的能力 ;采用电镜技术观察感染细胞的超微结构病变 ;采用 FITC- Annexin V/PI荧光标记流式细胞术 ,检测紫外线灭活前后的上述钩体菌株诱导 Vero和 J774 A.1细胞凋亡或坏死的情况。结果 :问号钩体 5 6 6 0 1和 5 6 6 0 8株对 Vero细胞的黏附率分别为 2 4 .2 %和 2 2 .9% (P>0 .0 5 ) ;对 J774 A.1细胞的黏附率分别为4 9.0 %和 4 6 .9% (P>0 .0 5 ) ;双曲钩体 Patoc 株不能黏附细胞。两株问号钩体侵入细胞后形成含有钩体的特殊吞噬泡 ,并引起相似的细胞超微结构病变 ,如细胞核染色质浓缩或溶解、细胞空泡变性、线粒体肿胀及线粒体嵴消失、内质网肿胀和膜旁核糖体消失等。灭活前的 5 6 6 0 1和 5 6 6 0 8株引起的 Vero细胞凋亡率分别为 84 .4 %和 82 .8% ,灭活后则分别为 77.9%和 86 .1%。灭活前后的 5 6 6 0 1株均以诱导 Vero细胞晚期凋亡为主 ,其凋亡率分别 6 8.0 %和5 2 .9%。灭活前后的 5 6 6 0 8株均以诱导 Vero细胞早期凋亡为主 ,其凋亡率分别为 6 4.1%和 5 0 .1%Objective: To determine the ability of adhering and invading to host cells and the related pathologic changes of Leptospira spp. with different virulence. Methods: A special Fontana silver staining method was deve loped to observe the ability of L.interrogans serogroup Icterohaemorrhagiae serovar lai strain 56601 and serogroup Pomona serovar pomona strain 56608,and L.biflexa serogroup Samaranga serovar patoc strain Patoc Ⅰ to adhere Vero and J774A.1 cells.Ultrastructural lesions of the infected cells were examined by electron microscopy.By using flow cytometry with fluorescein labeling of FITC Annexin V/PI,apoptosis and necrosis of the Vero and J774A.1 cells induced by the leptospiral strains before and after inactivation with ultraviolet treatment were detec ted, respectively. Results: The adhering rates of L.interrogans strains 56601 and 56608 to the Vero cells were 24 2% and 22 9% ( P >0 05),while to the J774A 1 cells were 49 0% and 46 9% ( P >0 05),respectively. L.biflexa strain Patoc Ⅰ did not adhere to these two host cells.After the two strains of L.interrogans invaded two different cell lines,the special phagosomes containing the Leptospira were formed and similar cell ultrastructural lesions,such as chromatilysis and chromatin condensation,vacuolar degeneration,mitochondrion swelling and mitochondrial crista disappearance,and endoplasmic reticulum swelling and paramembranous ribosome disappearance,were observed.The apoptosis rates in the Vero cells caused by the L.interrogans strains 56601 and 56608 before and after ultraviolet inactivation were 84 4%,82 8% and 77 9%,86 1%, respectively.The L.interrogans strain 56601 mainly induced the terminal apoptosis in Vero cells with the rates of 68 0% and 52 9% before or after inactivation, while the L.interrogans strain 56608 mainly induced the early apoptosis in Vero cells with apoptosis rates of 64 1% and 50 1% before or after inactivation.In the J774A.1 cells,the L.inter

关 键 词：钩端螺旋体 问号/致病力 细胞黏附 胞吞作用 镀银染色法 Vero细胞/超微结构 J774A.1细胞/超微结构 凋亡 坏死 

分 类 号：R377[医药卫生—病原生物学] R514.4[医药卫生—基础医学]