Duan Xueqin, Li Huicong, Tan Xuewen, Liu Nishang, Wang Xingchen, Zhang Weimin, Liu Yingqiu, Ma Wuren, Wu Yi, Ma Lin, Fan Yunpeng
College of Veterinary Medicine, Northwest A&F University, Yangling 712100, PR China; Institute of Traditional Chinese Veterinary Medicine, Northwest A&F University , Yangling 712100, PR China; Agricultural Management Department, Sichuan Xuanhan Vocational Secondary School, Xuanhan 636350, PR China.
College of Veterinary Medicine, Northwest A&F University, Yangling 712100, PR China; Institute of Traditional Chinese Veterinary Medicine, Northwest A&F University , Yangling 712100, PR China.
Vet J. 2024 Apr;304:106083. doi: 10.1016/j.tvjl.2024.106083. Epub 2024 Feb 15.
Transmissible gastroenteritis virus (TGEV) is an important pathogen capable of altering the expression profile of cellular miRNA. In this study, the potential of Polygonum cillinerve polysaccharide (PCP) to treat TGEV-infected piglets was evaluated through in vivo experiments. High-throughput sequencing technology was employed to identify 9 up-regulated and 17 down-regulated miRNAs during PCP-mediated inhibition of TGEV infection in PK15 cells. Additionally, miR-181 was found to be associated with target genes of key proteins in the apoptosis pathway. PK15 cells were treated with various concentrations of PCP following transfection with miR-181 mimic or inhibitor. Real-time PCR assessed the impact on TGEV replication, while electron microscopy (TEM) and Hoechst fluorescence staining evaluated cellular functionality. Western blot analysis was utilized to assess the expression of key signaling factors-cytochrome C (cyt C), caspase 9, and P53-in the apoptotic signaling pathway. The results showed that compared with the control group, 250 μg/mL PCP significantly inhibited TGEV gRNA replication and gene N expression (P < 0.01). Microscopic examination revealed uniform cell morphology and fewer floating cells in PCP-treated groups (250 and 125 μg/mL). TEM analysis showed no typical virus structure in the 250 μg/mL PCP group, and apoptosis staining indicated a significant reduction in apoptotic cells at this concentration. Furthermore, PCP may inhibit TGEV-induced apoptosis via the Caspase-dependent mitochondrial pathway following miR-181 transfection. These findings provide a theoretical basis for further exploration into the mechanism of PCP's anti-TGEV properties.
传染性胃肠炎病毒(TGEV)是一种能够改变细胞微小RNA(miRNA)表达谱的重要病原体。在本研究中,通过体内实验评估了绵毛酸模叶蓼多糖(PCP)治疗TGEV感染仔猪的潜力。采用高通量测序技术鉴定了在PCP介导的PK15细胞中TGEV感染抑制过程中9个上调和17个下调的miRNA。此外,发现miR-181与凋亡途径中关键蛋白的靶基因相关。用miR-181模拟物或抑制剂转染后,用不同浓度的PCP处理PK15细胞。实时定量聚合酶链反应评估对TGEV复制的影响,而电子显微镜(TEM)和Hoechst荧光染色评估细胞功能。蛋白质免疫印迹分析用于评估凋亡信号通路中关键信号因子——细胞色素C(cyt C)、半胱天冬酶9和P53的表达。结果表明,与对照组相比,250μg/mL PCP显著抑制TGEV基因组RNA(gRNA)复制和基因N表达(P<0.01)。显微镜检查显示,PCP处理组(250和125μg/mL)细胞形态均匀,漂浮细胞较少。TEM分析显示,250μg/mL PCP组无典型病毒结构,此浓度下凋亡染色表明凋亡细胞显著减少。此外,miR-181转染后,PCP可能通过半胱天冬酶依赖性线粒体途径抑制TGEV诱导的凋亡。这些发现为进一步探究PCP抗TGEV特性的机制提供了理论依据。