组蛋白去乙酰化酶抑制剂对小反刍兽疫病毒复制的影响

doi:10.11843/j.issn.0366-6964.2022.07.027

摘要/Abstract

摘要： 旨在探究组蛋白去乙酰化酶抑制剂对小反刍兽疫病毒(peste des petits ruminants virus，PPRV)复制的影响，以确定组蛋白去乙酰化酶(histone deacetylases，HDACs)在PPRV复制过程中的作用。采用RT-qPCR法检测PPRV感染后Vero细胞HDACs(HDAC1~11) mRNA表达水平的变化；用针对不同类型HDACs的抑制剂处理感染PPRV的Vero细胞48 h，Western blot筛选影响PRPV N蛋白表达的抑制剂；应用筛选出的抑制剂处理感染PPRV的Vero细胞，通过RT-qPCR、Western blot和TCID₅₀法进一步分析抑制剂对病毒RNA、蛋白和病毒滴度的影响。结果显示，PPRV感染后，Vero细胞HDAC2 mRNA水平显著升高(P<0.05)；SAHA、TMP269、MGCD0103处理后，PPRV N蛋白的表达量明显降低，且呈剂量负相关；SAHA、TMP269、MGCD0103也显著降低PPRV N RNA表达水平(P<0.05，P<0.05，P<0.01)和病毒滴度(P<0.05,P<0.05,P<0.01)。这表明Ⅱa类HDACs抑制剂和Ⅰ类HDACs抑制剂能抑制PPRV的复制，Ⅰ类HDACs (HDAC1~3)和Ⅱa类HDACs (HDAC4~5、HDAC7、HDAC9)可能参与调控PPRV的感染。

关键词: 组蛋白去乙酰化酶抑制剂, 组蛋白去乙酰化酶, 小反刍兽疫病毒, 复制

Abstract: The present study aimed to explore the effect of histone deacetylase(HDAC) inhibitors on peste des petits ruminants virus (PPRV) replication, and to determine the role of HDACs involved in the PPRV replication.The mRNA expression of HDACs (HDAC1-11) was examined by RT-qPCR assay in PPRV-infected Vero cells. To screen the inhibitors that affect PPRV replication, the expression level of N protein was evaluated in infected Vero cells by Western blot following treatment with the various inhibitors for different types of HDACs for 48 h. The selected inhibitors were used to treat PPRV-infected Vero cells, and the effects of inhibitors on PPRV N RNA, N protein and virus titer were further analyzed by RT-qPCR, Western blot and TCID₅₀, respectively. The results showed that PPRV infection in Vero cells resulted in a significant increase of the mRNA expression level of HDAC2 (P<0.05). After treatment with SAHA, TMP269 and MGCD0103, the expression level of PPRV N protein was significantly reduced and negatively correlated with the doses of these inhibitors. SAHA, TMP269 and MGCD0103 also significantly reduced the expression level of PPRV N RNA (P<0.05, P<0.05, P<0.01) and virus titer (P<0.05, P<0.05, P<0.01), respectively. These results indicated that class IIa HDACs inhibitors and class I HDACs inhibitors significantly inhibited PPRV replication in Vero cells, suggesting that class I HDACs (HDAC1-3) and class IIa HDACs (HDAC4-5, HDAC7, HDAC9) may be involved in the regulation of PPRV infection.

Key words: histone deacetylase inhibitor, histone deacetylase, peste des petits ruminants virus, replication

中图分类号:

S852.659.5

潘春容, 邓瑞雪, 胡林杰, 朱学亮, 孙跃峰, 王桂荣, 蒙学莲. 组蛋白去乙酰化酶抑制剂对小反刍兽疫病毒复制的影响[J]. 畜牧兽医学报, 2022, 53(7): 2307-2316.

PAN Chunrong, DENG Ruixue, HU Linjie, ZHU Xueliang, SUN Yuefeng, WANG Guirong, MENG Xuelian. Effect of Histone Deacetylase Inhibitors on Peste Des Petits Ruminants Virus Replication[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53(7): 2307-2316.

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