Zhan Jiafei, Song Hongyu, Wang Ning, Guo Cheng, Shen Nengxing, Hua Ruiqi, Shi Yuan, Angel Christiana, Gu Xiaobin, Xie Yue, Lai Weimin, Peng Xuerong, Yang Guangyou
Department of Parasitology, College of Veterinary Medicine, Sichuan Agricultural University, Chengdu, China.
Department of Veterinary Parasitology, Faculty of Veterinary Sciences, Shaheed Benazir Bhutto University of Veterinary and Animal Sciences, Sakrand, Pakistan.
Front Microbiol. 2020 Apr 22;11:729. doi: 10.3389/fmicb.2020.00729. eCollection 2020.
The larval stage of sensu lato, resulting in cystic echinococcosis, a parasitic zoonosis, causes huge economic losses to the livestock industry and poses a threat to public health. Inhibitor of apoptosis proteins (IAPs) is a class of endogenous anti-apoptotic family, which plays a significant functional role in the regulation of organism's development. Herein, to explore potential functions of IAPs in , two members of IAPs from (Eg-IAP and Eg-BIRP) were cloned, expressed, and molecularly characterized. Eg-IAP and Eg-BIRP encoded putative 331 and 168 residue proteins, respectively. Bioinformatic analysis showed that both proteins contained a type II BIR domain-the essential functional domain of IAPs. Fluorescence immunohistochemistry revealed that both proteins were ubiquitously localized in all life-cycle stages of . Our fluorescent quantitative PCR (RT-qPCR) results revealed relatively higher transcription levels of two Eg-IAPs in protoscoleces (PSCs) compared to the 18-day strobilated worms. We further used different concentrations of LCL161, a Smac-mimetic pan-IAPs inhibitor, to induce the apoptosis in PSCs , and revealed that the survival rate of PSCs and transcription levels of both genes were negatively correlated with the concentration of LCL161. While the results of light microscopy, transmission electron microscopy (TEM), and terminal deoxynucleotidyl transferase dUTP nick end labeling (TUNEL) assay also showed a higher apoptotic rate in PSCs with the increasing concentrations of LCL161. Taken together, our findings provide the reasonable evidence that both Eg-IAP and Eg-BIRP have potential implication in critical anti-apoptotic roles during the development of .
狭义的幼虫阶段会导致囊型包虫病，这是一种寄生性人畜共患病，给畜牧业造成巨大经济损失，并对公众健康构成威胁。凋亡抑制蛋白（IAPs）是一类内源性抗凋亡家族，在生物体发育调控中发挥着重要功能作用。在此，为了探究IAPs在[具体物种]中的潜在功能，克隆、表达并对来自[具体物种]的IAPs的两个成员（Eg-IAP和Eg-BIRP）进行了分子特征分析。Eg-IAP和Eg-BIRP分别编码推定的331和168个残基的蛋白质。生物信息学分析表明，这两种蛋白质都含有II型BIR结构域——IAPs的关键功能结构域。荧光免疫组织化学显示，这两种蛋白质在[具体物种]的所有生命周期阶段均广泛分布。我们的荧光定量PCR（RT-qPCR）结果显示，与18天的成虫节片相比，原头蚴（PSCs）中两种Eg-IAPs的转录水平相对较高。我们进一步使用不同浓度的LCL161（一种Smac模拟泛IAPs抑制剂）诱导PSCs凋亡，结果显示PSCs的存活率和这两个基因的转录水平与LCL161的浓度呈负相关。同时，光学显微镜、透射电子显微镜（TEM）和末端脱氧核苷酸转移酶dUTP缺口末端标记（TUNEL）分析结果也显示，随着LCL161浓度的增加，PSCs的凋亡率更高。综上所述，我们的研究结果提供了合理证据，表明Eg-IAP和Eg-BIRP在[具体物种]发育过程中的关键抗凋亡作用中都具有潜在意义。
