Chu Jia-Qi, Jing Kai-Peng, Gao Xiang, Li Peng, Huang Rui, Niu Yan-Ru, Yan Shou-Quan, Kong Jun-Chao, Yu Cai-Yuan, Shi Ge, Fan Yi-Ming, Lee Young-Ha, Zhou Yu, Quan Juan-Hua
a Stem Cell Research and Cellular Therapy Center, Affiliated Hospital of Guangdong Medical University , Zhanjiang , China.
b Laboratory Institute of Minimally Invasive Orthopedic Surgery, Affiliated Hospital of Guangdong Medical University , Zhanjiang , China.
Cell Cycle. 2017 Mar 4;16(5):477-486. doi: 10.1080/15384101.2017.1281484. Epub 2017 Jan 23.
Autophagy and apoptosis are critical for controlling Toxoplasma gondii (T. gondii) infection. T. gondii infection during pregnancy can damage the fetus and cause birth defects; however, the molecular mechanisms of this process are poorly understood. This study aims to determine the activities of autophagy and apoptosis as well as their regulatory mechanisms during T. gondii infection by using human umbilical cord mesenchymal stem cells (hUC-MSCs) as a model of congenital diseases. LC3B, a hallmark protein of autophagy was incrementally upregulated with the infection duration, whereas p62 was downregulated in T. gondii-infected hUC-MSCs. Concurrent to this result, the invasion of T. gondii into hUC-MSCs increased in a time-dependent manner. The expression levels of Bcl-2 family proteins including Bcl-2, Bcl-xL, Bim, Bax, Bid and Bak were not altered; however, Mcl-1 levels in hUC-MSCs were dramatically decreased upon T. gondii infection. In addition, at 24 h post-infection, cleaved PARP and cleaved caspase-3 protein levels were elevated in hUC-MSCs. Importantly, Mcl-1 overexpression reduced the levels of autophagy- and apoptosis-related proteins in T. gondii-infected hUC-MSCs. Mcl-1 proteins were primarily expressed in the fraction containing mitochondria and strongly interacted with Beclin-1 under normal conditions; however, these interactions were remarkably attenuated by T. gondii infection. These results suggest that mitochondrial Mcl-1 is an essential signaling mediator regulating the activation of autophagy and apoptosis during T. gondii infection.
自噬和凋亡对于控制弓形虫感染至关重要。孕期弓形虫感染会损害胎儿并导致出生缺陷;然而,这一过程的分子机制尚不清楚。本研究旨在以人脐带间充质干细胞(hUC-MSCs)作为先天性疾病模型,确定弓形虫感染期间自噬和凋亡的活性及其调控机制。自噬的标志性蛋白LC3B随着感染时间的延长而逐渐上调,而在弓形虫感染的hUC-MSCs中p62下调。与此结果一致,弓形虫对hUC-MSCs的侵袭呈时间依赖性增加。包括Bcl-2、Bcl-xL、Bim、Bax、Bid和Bak在内的Bcl-2家族蛋白的表达水平没有改变;然而,弓形虫感染后hUC-MSCs中的Mcl-1水平显著降低。此外,在感染后24小时,hUC-MSCs中裂解的PARP和裂解的caspase-3蛋白水平升高。重要的是,Mcl-1过表达降低了弓形虫感染的hUC-MSCs中自噬和凋亡相关蛋白的水平。Mcl-1蛋白主要在线粒体组分中表达,在正常条件下与Beclin-1强烈相互作用;然而,弓形虫感染显著减弱了这些相互作用。这些结果表明,线粒体Mcl-1是调节弓形虫感染期间自噬和凋亡激活的重要信号介质。