堆型艾美耳球虫裂殖子外分泌蛋白对细胞凋亡的抑制作用

Inhibitory Effect of Eimeria acervulina Merozoites Exocrine Proteins on Host Cell Apoptosis

为了研究堆型艾美耳球虫(Eimeria acervulina)裂殖子的外分泌蛋白对马-达氏牛肾(MDBK)细胞凋亡的抑制作用,本试验建立Transwell小室间接共培养体系,即将E.acervulina裂殖子与MDBK细胞共培养,采用流式细胞术分别检测共培养1.5、3和6 h MDBK细胞的凋亡情况,利用液相色谱串联质谱(LC-MS/MS)对共培养6 h后的细胞上清液中的成分进行鉴定。结果显示,1.5、3和6 h共培养组MDBK细胞的总细胞凋亡率分别为(5.92±0.19)%、(8.13±0.32)%和(11.57±0.37)%,与对照组相比,总细胞凋亡率均极显著降低(P<0.01)。LC-MS/MS分析共鉴定到65个E.acervulina的外分泌蛋白,包括热休克蛋白、微线蛋白、能量代谢酶分子、蛋白磷酸酶分子和未知功能蛋白等。结果表明,E.acervulina裂殖子外分泌蛋白可抑制MDBK细胞凋亡,具体哪种蛋白在抑制细胞凋亡中发挥作用有待后续进一步研究。

To investigate the inhibitory effect of exocrine proteins from Eimeria acervulina merozoites on apoptosis of Madin-Darby bovine kidney(MDBK)cells,this study established a Transwell chamber indirect co-culture system,which involved co-culturing E.acervulina merozoites with MDBK cells.Flow cytometry was used to assess apoptosis in MDBK cells after 1.5,3,and 6 hours of co-culture,and liquid chromatography-tandem mass spectrometry(LC-MS/MS)was employed to identify components in the cell supernatant after 6 hours of co-culture.The results showed that the total apoptosis rates of MDBK cells in the co-culture groups at 1.5,3,and 6 hours were(5.92±0.19)%,(8.13±0.32)%,and(11.57±0.37)%,respectively.These rates were significantly lower compared with the control group(P<0.01).LC-MS/MS analysis identified 65 exocrine proteins from E.acervulina,including heat shock proteins,microneme proteins,energy metabolism enzymes,protein phosphatase molecules,and proteins with unknown functions.The results indicate that exocrine proteins from E.acervulina merozoites can inhibit apoptosis in MDBK cells,and further research is needed to identify the specific proteins involved in this anti-apoptotic activity.