Zhou Jie, Gan Xiaofeng, Wang Yongzhong, Zhang Xian, Ding Xiaojuan, Chen Lingzhi, Du Jian, Luo Qingli, Wang Teng, Shen Jilong, Yu Li
Department of Microbiology and Parasitology, Anhui Provincial Laboratory of Microbiology and Parasitology, Anhui Key Laboratory of Zoonoses, Anhui Medical University, Hefei, 230032, PR China.
Clinical Laboratory, People's Hospital of Huaibei, Huaibei, 235000, PR China.
Parasit Vectors. 2015 Feb 4;8:73. doi: 10.1186/s13071-015-0670-3.
Toxoplasma gondii, an obligate intracellular pathogen, has a strong affinity for the nervous system. TgCtwh3, a representative Chinese 1 Toxoplasma strain prevalent in China, has the polymorphic features of the effectors ROP16I/III with type I and GRA15II with type II Toxoplasma strains. The interaction of this atypical strain with host cells remains extremely elusive.
Using a transwell system, neural stem cells C17.2 were co-cultured with the tachyzoites of TgCtwh3 or standard type I RH strain. The apoptosis levels of C17.2 cells and the expression levels of related proteins in the endoplasmic reticulum stress (ERS)-mediated pathway were detected by flow cytometry and Western blotting.
The apoptosis level of C17.2 cells co-cultured with TgCtwh3 had a significant increase compared to the negative control group; however, the apoptosis level in the TgCtwh3 group was significantly lower than that in the RH co-culture group. Western blotting analyses reveal that, after the C17.2 cells were co-cultured with TgCtwh3 and RH tachyzoites, the expression levels of caspase-12, CHOP and p-JNK in the cells increased significantly when compared to the control groups. After the pretreatment of Z-ATAD-FMK, an inhibitor of caspase-12, the apoptosis level of the C17.2 cells co-cultured with TgCtwh3 or RH tachyzoites had an apparent decline, and correspondingly, the expression levels of those related proteins were notably decreased.
Our findings suggest that TgCtwh3 may induce the apoptosis of the C17.2 cells by up-regulation of caspase-12, CHOP, and p-JNK, which are associated with ERS signaling pathways. This work contributes to better understanding the possible mechanism of brain pathology induced by T. gondii Chinese 1 isolates prevalent in China, and also reveals the potential value of ERS inhibitors to treat such related diseases in the future.
刚地弓形虫是一种专性细胞内病原体,对神经系统具有很强的亲和力。TgCtwh3是在中国流行的代表性中国1型弓形虫菌株,具有I型效应子ROP16I/III和II型弓形虫菌株GRA15II的多态性特征。这种非典型菌株与宿主细胞的相互作用仍然极其难以捉摸。
使用Transwell系统,将神经干细胞C17.2与TgCtwh3或标准I型RH菌株的速殖子共培养。通过流式细胞术和蛋白质印迹法检测C17.2细胞的凋亡水平以及内质网应激(ERS)介导途径中相关蛋白的表达水平。
与阴性对照组相比,与TgCtwh3共培养的C17.2细胞凋亡水平显著增加;然而,TgCtwh3组的凋亡水平显著低于RH共培养组。蛋白质印迹分析表明,C17.2细胞与TgCtwh3和RH速殖子共培养后,与对照组相比,细胞中caspase-12、CHOP和p-JNK的表达水平显著增加。在用caspase-12抑制剂Z-ATAD-FMK预处理后,与TgCtwh3或RH速殖子共培养的C17.2细胞凋亡水平明显下降,相应地,那些相关蛋白的表达水平也显著降低。
我们的研究结果表明,TgCtwh3可能通过上调与ERS信号通路相关的caspase-12、CHOP和p-JNK来诱导C17.2细胞凋亡。这项工作有助于更好地理解中国流行的刚地弓形虫中国1型分离株诱导脑部病变的可能机制,也揭示了ERS抑制剂在未来治疗此类相关疾病中的潜在价值。