微小隐孢子虫感染诱导HCT-8细胞凋亡规律的研究

Study on the rule of HCT-8 cell apoptosis induced by Cryptosporidium parvum infection

为了解微小隐孢子虫(Cryptosporidium parvum)感染诱导人结肠癌细胞(HCT-8细胞)凋亡规律,试验将HCT-8细胞分为感染组和对照组,感染组HCT-8细胞加入微小隐孢子虫子孢子(细胞和子孢子数量比为1∶20),对照组HCT-8细胞正常培养,同时加入青霉素与链霉素防止污染,在感染后0.5,4,8,12,24,36,48,72 h分别取样,采用Annexin V/PI双染色法对HCT-8细胞的凋亡情况进行检测。结果表明:感染后0.5,4,8,12,24,36,48,72 h,试验组细胞凋亡占比极显著高于对照组(P<0.001)。在微小隐孢子虫感染HCT-8细胞12 h前有一个明显抑制细胞凋亡的过程,凋亡细胞明显减少;感染12 h后凋亡细胞极显著增加(P<0.001);在感染后48 h细胞凋亡达到高峰。说明微小隐孢子虫感染后存在调节HCT-8细胞凋亡的机制,即感染早期抑制细胞凋亡以利于虫体发育,而感染后期促进细胞凋亡便于释放虫体扩大感染。

In order to understand the rule of human colon cancer cell(HCT-8 cell)apoptosis induced by Cryptosporidium parvum infection,HCT-8 cells was divided into two groups in this experiment,namely the infection group and the control group;in the infection group,HCT-8 cells were infected with Cryptosporidium parvum sporozoites(the ratio of cells to sporozoites was 1∶20),and in the control group,HCT-8 cells were cultured normally,and penicillin-streptomycin were added to prevent contamination.The samples were taken at 0.5,4,8,12,24,36,48,72 h after infection,and the apoptosis of HCT-8 cells was detected by Annexin V/PI double staining.The results showed that 0.5,4,8,12,24,36,48,72 h after infection,the percentage of cell apoptosis of the infection group was significantly higher than that of the control group(P<0.001).Twelve hours before infection with Cryptosporidium parvum,there was an obvious process of inhibiting cell apoptosis,and apoptotic cells were significantly reduced;12 hours after infection,apoptotic cells were significantly increased(P<0.001),and cell apoptosis reached a peak at 48 h after infection.The results suggested that after Cryptosporidium parvum infection,there was a mechanism of regulating HCT-8 cell apoptosis,which meant that it inhibited apoptosis in the early stage of infection to facilitate the development of the parasite,and promoted the apoptosis in the later stage to facilitate the release of the parasite and to expand the infection.