创伤弧菌诱导小鼠树突状细胞DC2.4凋亡的实验研究

Experimental Research on the Induction of Apoptosis in Mouse Dendritic Cells by Vibrio Vulnificus

通过建立Vv1.1758标准菌株与小鼠树突状细胞DC 2.4的细胞感染模型,计算其感染率,并采用FITC-Annexin V/PI荧光标记流式细胞术检测小鼠树突状细胞DC 2.4的凋亡情况。结果显示:Vv1.1758标准菌株与小鼠树突状细胞DC 2.4混合培养1h后,即见有创伤弧菌的侵入,混合培养2h、3h、4h、5h、6h后,细菌侵入细胞现象明显,侵入率分别为(36.3±3.4)%、(55.1±4.7)%、(67.9±6.3)%、(83.9±5.8)%和(91.9±8.3)%,与混合培养1h(12.1±1.3)比较,细菌侵入率明显升高(P<0.05);Vv 1.1758标准菌株与小鼠树突状细胞DC 2.4混合培养1h,2h,3h,4h,5h,6h后,细胞凋亡率分别为(6.2±1.1)%、(26.9±8.8)%、(35.5±10.2)%、(43.3±11.7)%、(60.8±13.8)%和(76.1±15.3)%,由此得出,创伤弧菌可侵入DC 2.4细胞,诱导细胞凋亡可能是其损伤DC 2.4的主要机制。

Objective： To investigate the induction of apoptosis in mouse dendritic cells DC 2.4 by Vibrio vulnificus. Methods： V. vulnificusl. 1758 strain was co--cultured with DC 2.4 cells and the rates of invasion were calculated. The rates of apoptosis on mouse dendritic cells DC 2.4 were detected by the FITC -- Annexin V/PI fluorescence marker flow eytometry. Results： There were bacterial invasion while DC 2.4 cells co-- cultured with V. vulnifieusl. 1758 strain after 1 h and the rates of invasion were（36.3± 3.4）%, （55.1 ± 4.7）%,（67.9±6.3）%,（83.9±5.8）% and（91.9±8.3）% at 2h,3h,4h,5h,and 6h respectively. The rates of invasion rose rapidly at at l h co--cultured （12.1 ± 1.3）% （P〈0.05）. The rates of apoptosis on mouse dendritic cells DC 2.4 were （6.2±1.1）%,（26.9±8.8）%,（35.5±10.2）%,（43.3± 11.7）%,（60.8±13.8）% and （76.1± 15.3）% at 1 h,2 h,3 h,4 h,5 h and 6 h respectively. Conclusion： Vibrio vulnificus 1. 1758 strain has an ability to enter DC 2.4 cells and causes injury of DC 2.4 cells via inducing apoptosis, which may be the major pathogenic mechanism of vibrio vulnificus.