甘露消毒丹对猫传染性腹膜炎病毒复制及细胞自噬功能的影响

doi:10.16431/j.cnki.1671-7236.2025.11.045

摘要/Abstract

摘要： 【目的】猫传染性腹膜炎(feline infectious peritonitis,FIP)由猫传染性腹膜炎病毒(Feline infectious peritonitis virus,FIPV)引起，严重危害猫的健康和生命，当前尚无安全有效的抗病毒药物治疗该病。本研究旨在探究FIPV感染宿主细胞后，甘露消毒丹对病毒复制的影响，以期揭示相关中药治疗作用机制，为FIP的治疗积累资料。【方法】通过Western blotting检测FIP患猫肠道组织自噬相关蛋白LC3、Beclin-1和p62表达情况；在体外将FIPV毒株B5接种于猫肾细胞(CRFK)，观察其对细胞形态和生长状态的影响；利用细胞毒性试验筛选对CRFK细胞生长无毒副作用的甘露消毒丹浓度，并在该药物浓度下，利用实时荧光定量PCR检测细胞内病毒载量；通过Western blotting验证病毒复制过程中甘露消毒丹对LC3蛋白及MAPK通路相关蛋白MEK、ERK、mTOR的影响。【结果】与对照组相比,在FIP患猫肠道组织中，FIPV可显著降低LC3、Beclin-1和p62的表达水平及LC3-Ⅱ/LC3-Ⅰ比值(P＜0.05)；在CRFK细胞中，FIPV感染诱导细胞发生脱落、融合；与病毒组相比，80 μg/mL甘露消毒丹治疗后可显著降低细胞内病毒载量，升高LC3-Ⅱ/LC3-Ⅰ比值，下调MEK蛋白表达，上调EPK和mTOR蛋白表达(P<0.05)。【结论】甘露消毒丹通过调控MAPK通路，提高细胞自噬水平进而清除FIPV，达到抑制病毒复制的效果，逆转了FIPV感染导致的宿主免疫力降低现象。

关键词: 甘露消毒丹; 猫传染性腹膜炎病毒(FIPV); 自噬; MAPK通路

Abstract: 【Objective】 Feline infectious peritonitis (FIP) was caused by Feline infectious peritonitis virus (FIPV),which seriously endangered the health and life of cats.At present,there were no safe and effective antiviral drugs for the treatment of this disease.The aim of this study was to investigate the effect of Ganlu Xiaodu micropills on virus replication after FIPV infection of host cells,in order to reveal the mechanism of Chinese medicine treatment and accumulate data for the treatment of FIP. 【Method】 Western blotting was used to detect the expression of autophagy related proteins LC3,Beclin-1 and p62 in the intestinal tissues of cats with FIP.FIPV strain B5 was inoculated into CRFK cells in vitro to observe its effect on cell morphology and growth status.The concentration of Ganlu Xiaodu micropills,which had no toxic and side effects on the growth of CRFK cells,was screened by cytotoxicity test.At this concentration,the intracellular viral load was detected by Real-time quantitative PCR.Western blotting was used to verify the effect of Ganlu Xiaodu micropills on protein LC3，and the MAPK pathway upstream of autophagy related proteins MEK,ERK and mTOR during virus replication. 【Result】 Compared with control group,in the intestinal tissues of cats with FIP,FIPV could significantly reduce the expression levels of LC3,Beclin-1 and p62,as well as the LC3-Ⅱ/LC3-Ⅰ ratio (P＜0.05).In CRFK cells,FIPV infection induced cell shedding and fusion.Compared with virus group,80 μg/mL Ganlu Xiaodu micropills could significantly reduce the intracellular viral load,increase the ratio of LC3-Ⅱ/LC3-Ⅰ,downregulate the expression of MEK protein,and upregulate the expressions of EPK and mTOR proteins (P<0.05). 【Conclusion】 Ganlu Xiaodu micropills could inhibit virus replication by regulating MAPK pathway,improving the level of autophagy and eliminating FIPV,reversing the reduction of host immunity caused by FIPV infection.

Key words: Ganlu Xiaodu micropills; Feline infectious peritonitis virus (FIPV); autophagy; MAPK pathway

中图分类号:

殷桂彬, 郭昊彤, 赵文豪, 宋健, 羊雨恬, 李佳, 刘晓晔, 陈武. 甘露消毒丹对猫传染性腹膜炎病毒复制及细胞自噬功能的影响[J]. 中国畜牧兽医, 2025, 52(11): 5510-5519.

YIN Guibin, GUO Haotong, ZHAO Wenhao, SONG Jian, YANG Yutian, LI Jia, LIU Xiaoye, CHEN Wu. Effect of Ganlu Xiaodu Micropills on Feline Infectious Peritonitis Virus Replication and Cellular Autophagic Function[J]. China Animal Husbandry & Veterinary Medicine, 2025, 52(11): 5510-5519.

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