Polygonum Cillinerve polysaccharide inhibits Transmissible gastroenteritis virus by regulating microRNA-181
Duan, X.; Li, H.; Tan, X.; Liu, N.; Wang, X.; Zhang, W.; Liu, Y.; Ma, W.; Wu, Y.; Ma, L.; Fan, Y.
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Transmissible gastroenteritis virus (TGEV) is an important pathogen that can cause changes in the expression profile of cell miRNA. In this study, PCP could treat piglets infected with TGEV by in vivo experiment. High-throughput sequencing technology was used to detect that 9 miRNAs were up-regulated and 17 miRNAs were down-regulated during PCP inhibition of TGEV infection in PK15 cells. Meanwhile, miR-181 was related to the target genes of key proteins of apoptosis pathway. PK15 was treated with different PCP after transfection of miR-181 mimic or inhibitor. Real-time PCR was used to detect the effect of TGEV replication, electron microscopy, TEM and Hoechst fluorescence staining were used to detect the biological function of the treated cells, and western blot (WB) was used to detect the expression of key signaling factors cyt C, caspase 9 and P53 in the apoptotic signaling pathway. The results showed that compared with the control group, 250 g/mL PCP significantly inhibited the replication of TGEV gRNA and gene N (P < 0.01). PK15 cells in PCP groups (250 and 125 g/mL) had uniform cell morphology and a small number of floating cells by microscopic, no typical virus structure was observed in 250 g/mL PCP group under TEM, and apoptosis staining showed that 250 g/mL PCP significantly reduced the number of apoptotic cells. PCP may inhibit TGEV-induced apoptosis through Caspase-dependent mitochondrial pathway after transfection of miR-181. The above results provided a theoretical basis for further research on the mechanism of PCP anti-TGEV.
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