Differences in the secretory exosomes of Clonorchis sinensis adults at different incubation times.

Exosomes are small membrane vesicles of endocytic origin and widely involved in a variety of physiological and pathological conditions. Exosome-like vesicles (ELVs) have been identified to mediate the parasite-host interactions and communication. Thus, increased knowledge of C. sinensis ELVs could provide insights into parasite-host interactions. In this experiment, ELVs was purified by ultracentrifugation from the culture medium of C. sinensis adults in vitro incubated for 24 h and 48 h, respectively. Transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA) confirmed that the purified vesicles which ranged from 30 to 150 nm in size were present in the culture medium. Small RNA high-throughput sequencing analysis identified 51 miRNAs, including 37 known C. sinensis miRNAs, 3 novel C. sinensis miRNAs and 11 rat miRNAs. The sequencing data were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). The biological function of targets of known C. sinensis miRNAs were proved to associated with signal transduction, infectious diseases and the immune system. Further, 15 miRNAs were classified as differentially expressed in the 24h-ELVs compared to the 48h-ELVs. We found that the numbers and expression levels of most miRNAs from 24h-ELVs were more and higer than 48h-ELVs'. Our work provides important data for understanding the molecular mechanisms underlying the pathogenesis of C. sinensis adults ELVs.