摘要
颗粒蛋白前体(PGRN)是一种分泌型多功能蛋白,具有多种生物学功能,在炎症反应、创伤修复、溶酶体功能、自噬、神经发育等多种生命活动过程中发挥重要作用。课题组前期研究结果证实,ORFV能够诱导OFTu细胞发生自噬,作为细胞自噬的重要调控分子,PGRN是否参与对ORFV诱导OFTu细胞自噬的调控尚不清楚。本研究首先从转录和蛋白水平上分析PGRN在ORFV感染细胞中的表达变化规律,结果显示,PGRN发生显著上调的时间与ORFV诱导自噬发生的时间一致。进一步研究显示,敲低PGRN能够抑制LC3的型别转换,并使P62表达上调;而体外添加PGRN重组蛋白则促进LC3Ⅰ向LC3Ⅱ的转换,P62表达下调。以上结果表明,PGRN参与对ORFV诱导细胞自噬的调控。此外,病毒噬斑试验结果显示,PGRN能够促进ORFV复制,且可解除自噬抑制剂对病毒复制的抑制,该结果表明,PGRN能够通过调节ORFV激活的自噬影响病毒的复制。本研究结果将为明确PGRN对ORFV诱导自噬的调控作用及进一步揭示ORFV致病机制奠定基础。
Progranulin(PGRN)is a secreted multifunctional protein with multiple biological functions,which plays an important role in many activities such as inflammatory response,wound repair,lysosomal function,autophagy,and neurodevelopment.Previous studies confirmed that ORFV induces autophagy in OFTu cells.However,whether PGRN,an important regulatory molecule of autophagy,is involved in the regulation of ORFV-induced autophagy in OFTu cells is not clear.In view of this,we first analyzed the expression pattern of PGRN in ORFV-infected cells from the transcriptional and translational levels,It was found that the expression pattern of PGRN was consistent with the pattern of ORFV-induced autophagy.Further study showed that knockdown of PGRN expression inhibited the type conversion of LC3and up-regulated the P62expression,while addition of PGRN recombinant protein promoted the conversion of LC3Ⅰto LC3Ⅱ,and the downregulated the P62expression.These results indicated that PGRN was involved in the regulation of ORFV-induced autophagy.Furthermore,viral plaque assay showed that PGRN promoted the replication of ORFV,while reversed the inhibition of viral replication by autophagy inhibitor,indicating that PGRN affects the replication of the virus by regulating ORFV-activated autophagy.The results of this study are conducive to clarifying the regulatory effect of PGRN on ORFV-induced autophagy and lay a foundation for further investigation on the pathogenic mechanism of ORFV.
作者
甄瑞雪
吕丽君
陆慧君
刘兴源
徐梦实
关继羽
贺文琦
高丰
赵魁
ZHEN Ruixue;LYU Lijun;LU Huijun;LIU Xingyuan;XU Mengshi;GUAN Jiyu;HE Wenqi;GAO Feng;ZHAO Kui(Key Laboratory of Zoonosis Research,Ministry of Education,College of Veterinary Medicine,Jilin University,Changchun 130062,China)
出处
《中国兽医学报》
CAS
CSCD
北大核心
2023年第5期908-914,共7页
Chinese Journal of Veterinary Science
基金
吉林省重点科技攻关资助项目(20210202042NC)
