改良育阴方对非洲猪瘟病毒感染PAMs的cGAS-STING通路影响

doi:10.11843/j.issn.0366-6964.2024.12.045

摘要/Abstract

摘要：

旨在为评价中药体外对非洲猪瘟病毒的抑制作用，本研究将5种试验用药：连翘、马勃、改良育阴方(modified Yuyin decoction, MYY)、复方鱼腥草(复方2)和银翘马勃散(复方3)，通过qPCR检测不同中药在被非洲猪瘟病毒(African swine fever virus, ASFV)感染的猪肺泡巨噬细胞(porcine alveolar macrophages, PAMs)中，对ASFV编码的衣壳蛋白p72的影响，筛选出可以显著降低ASFV-p72基因表达的中药为改良育阴方(MYY)。采用LC-MS分析检测出改良育阴方的主要化学成分；采用CCK8法观察改良育阴方对猪肺泡巨噬细胞(PAMs)活力的影响；荧光定量PCR检测ASFV-p72基因的表达；Western blot和ELISA检测CGAS-STING信号通路蛋白表达水平；RU.521作为cGAS抑制剂用于验证MYY在cGAS-STING通路中对ASFV的作用。结果表明，在PAMs中，MYY可以显著降低ASFV-p72基因表达，在ASFV感染2 h给药效果最佳，对ASFV感染PAMs后的细胞活力具有浓度依赖性改善作用。攻毒后2 h，使用MYY发现PAMs细胞上清液中干扰素基因刺激蛋白(STING)、TANK结合激酶1(TBK1)、干扰素调节因子3(IRF3)、干扰素诱导跨膜蛋白3(IFITM3)表达量的下降，PAMs细胞内环状GMP-AMP合成酶(cGAS)、干扰素β(IFNβ)蛋白量表达增加，MYY可激活cGAS-STING-TBK1-IRF3-IFNβ信号通路，促进IFITM3的表达。经RU.521处理后，MYY仍能提高细胞活力，降低ASFV-p72基因的表达。综上所述，MYY具有抑制ASFV的潜在作用，可能与cGAS-STING信号通路的激活促进IFITM3的表达有关。

关键词: 改良育阴方, 非洲猪瘟病毒, ASFV-p72基因, CGAS-STING信号通路, LC-MS分析

Abstract:

In order to evaluate the efficacy of Chinese traditional medicine against African swine fever virus (ASFV) in vitro, five experimental drugs were used in this study. The effects of different Chinese herbal medicines on ASFV encored capshell protein p72 in porcine alveolar macrophages (PAMs) infected with African swine fever virus were detected by qPCR. Modified Yuyin decoction (MYY) was selected as a Chinese medicine that could significantly reduce the expression of ASFV-p72 gene. The main components of anti-ASFV were detected by LC-MS analysis. CCK8 method was used to observe the effect of modified Yuyin decoction on the activity of PAMs. The expression of ASFV-p72 gene was detected by fluorescence quantitative PCR. Western blot and ELISA were used to detect CGAS-STING signaling pathway protein expression. RU.521 was used as a cGAS inhibitor to verify the anti-ASFV effect of MYY in the CGAS-STING pathway. The results showed that MYY can significantly reduce the expression of ASFV-p72 gene in PAMs, and the best effect is given at 2 h after ASFV infection, and it has a concentration-dependent improvement effect on the cell viability after ASFV infection of PAMs. The expression levels of interferon gene stimulating protein (STING), TANK binding kinase 1 (TBK1), interferon regulatory factor 3 (IRF3) and interferon induced transmembrane protein 3 (IFITM3) in the supernatant of PAMs cells were decreased by MYY 2 h after the challenge. The expression of cyclic GMP-AMP synthetase (cGAS) and interferon β (IFNβ) protein in PAMs cells increased, and MYY could activate the cGAS-STING-TBK1-IRF3-IFNβsignaling pathway and promote the expression of IFITM3. After treatment with RU.521, MYY could still increase cell viability and decrease the expression of ASFV-p72 gene. In summary, MYY has a potential anti-ASFV effect, which may be related to the activation of cGAS-STING signaling pathway to promote the expression of IFITM3.

Key words: modified Yuyin decoction, African swine fever virus, ASFV-p72 gene, CGAS-STING signaling pathway, LC-MS analysis

中图分类号:

陈晓丽, 周佳浩, 周静, 屈倩, 王志华, 熊鹰, 朱咏琪, 贾伟新, 吕伟杰, 郭世宁. 改良育阴方对非洲猪瘟病毒感染PAMs的cGAS-STING通路影响[J]. 畜牧兽医学报, 2024, 55(12): 5839-5853.

CHEN Xiaoli, ZHOU Jiahao, ZHOU Jing, QU Qian, WANG Zhihua, XIONG Ying, ZHU Yongqi, JIA Weixin, LÜ Weijie, GUO Shining. Effect of Modified Yuyin Decoction on cGAS-STING Pathway of African Swine Fever Virus Infected PAMs[J]. Acta Veterinaria et Zootechnica Sinica, 2024, 55(12): 5839-5853.

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参考文献 40

1	DIXON L K , SUN H , ROBERTS H . African swine fever[J]. Antiviral Res, 2019, 165, 34- 41. doi: 10.1016/j.antiviral.2019.02.018
2	陈画菡, 朱君海, 高飞, 等. 非洲猪瘟检测方法研究进展[J]. 中国兽医杂志, 2024, 60 (5): 91- 100.
	CHEN H H , ZHU J H , GAO F , et al. Research progress of detection methods for African swine fever[J]. Chinese Journal of Veterinary Medicine, 2024, 60 (5): 91- 100.
3	ARABYAN E , HAKOBYAN A , KOTSINYAN A , et al. Genistein inhibits African swine fever virus replication in vitro by disrupting viral DNA synthesis[J]. Antiviral Res, 2018, 156, 128- 137. doi: 10.1016/j.antiviral.2018.06.014
4	方建国, 施春阳, 汤杰, 等. 大青叶抗内毒素活性部位筛选[J]. 中草药, 2004, 35 (1): 60- 62. doi: 10.3321/j.issn:0253-2670.2004.01.026
	FANG J G , SHI C Y , TANG J , et al. Screening of active fraction of antiendotoxin from Folium Isatidis[J]. Chinese Traditional and Herbal Drugs, 2004, 35 (1): 60- 62. doi: 10.3321/j.issn:0253-2670.2004.01.026
5	黄楠, 张兵, 冯燕, 等. 白及提取物小鼠体内抗流感病毒药效研究[J]. 浙江中医药大学学报, 2019, 43 (8): 734- 742.
	HUANG N , ZHANG B , FENG Y , et al. Study on the anti-influenza virus efficacy of the Bletilla striata extracts in mice[J]. Journal of Zhejiang Chinese Medical University, 2019, 43 (8): 734- 742.
6	梁怡平, 钟秀会. 中药抗流感病毒的机理与应用前景[J]. 今日畜牧兽医, 2014, (1): 13- 15. doi: 10.3969/j.issn.1673-4092.2014.01.005
	LIANG Y P , ZHONG X H . Mechanism and application prospect of Chinese medicine against influenza virus[J]. Animal Husbandry and Veterinary Today, 2014, (1): 13- 15. doi: 10.3969/j.issn.1673-4092.2014.01.005
7	徐培平, 赵昉, 刘妮, 等. 中药体外防治甲型H1N1流感病毒实验研究[J]. 实用预防医学, 2012, 19 (8): 1121- 1124. doi: 10.3969/j.issn.1006-3110.2012.08.001
	XU P P , ZHAO F , LIU N , et al. In vitro study on preventive effects of three Chinese medicines against influenza A (H1N1)[J]. Practical Preventive Medicine, 2012, 19 (8): 1121- 1124. doi: 10.3969/j.issn.1006-3110.2012.08.001
8	刘廷, 王海丹, 狄留庆, 等. 基于甲型H1N1流感病毒致MDCK细胞损伤保护作用的双黄连组方HPLC特征图谱谱效关系研究[J]. 中国中药杂志, 2015, 40 (21): 4194- 4199.
	LIU T , WANG H D , DI L Q , et al. HPLC specific chromatogram spectrum-effect relationship for Shuanghuanglian on MDCK cell injury induced by influenza A virus (H1N1)[J]. China Journal of Chinese Materia Medica, 2015, 40 (21): 4194- 4199.
9	宋晓东, 刘哲, 陈敬洲, 等. 体外筛选具有抗柯萨奇病毒作用的中药提取物[J]. 中国药物与临床, 2010, 10 (9): 974- 976. doi: 10.3969/j.issn.1671-2560.2010.09.003
	SONG X D , LIU Z , CHEN J Z , et al. Screening of traditional Chinese herbal extracts against Coxsackie virus in vitro[J]. Chinese Remedies & Clinics, 2010, 10 (9): 974- 976. doi: 10.3969/j.issn.1671-2560.2010.09.003
10	卫文峰, 张国成, 许东亮, 等. 柴胡黄芩炙甘草对小鼠CVB3心肌炎治疗作用的研究[J]. 中国当代儿科杂志, 2003, 5 (3): 223- 225. doi: 10.3969/j.issn.1008-8830.2003.03.010
	WEI W F , ZHANG G C , XU D L , et al. Therapeutic effects of bupleurum root, scutellaria root and baked licorice on viral myocarditis caused by coxsackievirus B3 in mice[J]. Chinese Journal of Contemporary Pediatrics, 2003, 5 (3): 223- 225. doi: 10.3969/j.issn.1008-8830.2003.03.010
11	HAKOBYAN A , ARABYAN E , KOTSINYAN A , et al. Inhibition of African swine fever virus infection by genkwanin[J]. Antiviral Res, 2019, 167, 78- 82. doi: 10.1016/j.antiviral.2019.04.008
12	李艳青, 朱秀高, 王晓田. 含金银花复方中药提取物方剂在非洲猪瘟防控中的作用[J]. 今日养猪业, 2021, (5): 88- 89.
	LI Y Q , ZHU X G , WANG X T . The role of Chinese herbal extract formula containing honeysuckle flower in the prevention and control of African swine fever[J]. Including Today's Pig Industry, 2021, (5): 88- 89.
13	毛世香, 黄金凤, 张方武, 等. 非洲猪瘟的卫气营血辨证及防控[J]. 畜禽业, 2021, 32 (3): 97- 98.
	MAO S X , HUANG J F , ZHANG F W , et al. Differentiation of Weiqi Ying blood syndrome and prevention and control of African swine fever[J]. Livestock and Poultry Industry, 2021, 32 (3): 97- 98.
14	LIU Y J , ZHANG X H , LIU Z X , et al. Toosendanin suppresses African swine fever virus replication through upregulating interferon regulatory factor 1 in porcine alveolar macrophage cultures[J]. Front Microbiol, 2022, 13, 970501. doi: 10.3389/fmicb.2022.970501
15	赵旭阳, 靳家鑫, 路闻龙, 等. 非洲猪瘟病毒免疫逃逸分子机制研究进展[J]. 畜牧兽医学报, 2022, 53 (7): 2074- 2082. doi: 10.11843/j.issn.0366-6964.2022.07.005
	ZHAO X Y , JIN J X , LU W L , et al. Advances in the molecular mechanism of immune escape of African swine fever virus[J]. Acta Veterinaria et Zootechnica Sinica, 2022, 53 (7): 2074- 2082. doi: 10.11843/j.issn.0366-6964.2022.07.005
16	GARCÍA-BELMONTE R , PÉREZ-NÚÑEZ D , PITTAU M , et al. African swine fever virus Armenia/07 virulent strain controls interferon beta production through the cGAS-STING pathway[J]. J Virol, 2019, 93 (12): e02298- 18.
17	RASMUSSEN J A , VILLUMSEN K R , ERNST M , et al. A multi-omics approach unravels metagenomic and metabolic alterations of a probiotic and synbiotic additive in rainbow trout (Oncorhynchus mykiss)[J]. Microbiome, 2022, 10 (1): 21. doi: 10.1186/s40168-021-01221-8
18	NAVARRO-REIG M , JAUMOT J , GARCÍA-REIRIZ A , et al. Evaluation of changes induced in rice metabolome by Cd and Cu exposure using LC-MS with XCMS and MCR-ALS data analysis strategies[J]. Anal Bioanal Chem, 2015, 407 (29): 8835- 8847. doi: 10.1007/s00216-015-9042-2
19	WISHART D S , TZUR D , KNOX C , et al. HMDB: the human metabolome database[J]. Nucl Acids Res, 2007, 35 (suppl_1): D521- D526.
20	HORAI H , ARITA M , KANAYA S , et al. MassBank: a public repository for sharing mass spectral data for life sciences[J]. J Mass Spectrom, 2010, 45 (7): 703- 714. doi: 10.1002/jms.1777
21	SUD M , FAHY E , COTTER D , et al. LMSD: LIPID MAPS structure database[J]. Nucl Acids Res, 2007, 35 (suppl_1): D527- D532.
22	ABDELRAZIG S , SAFO L , RANCE G A , et al. Metabolic characterisation of Magnetospirillum gryphiswaldense MSR-1 using LC-MS-based metabolite profiling[J]. RSC Adv, 2020, 10 (54): 32548- 32560. doi: 10.1039/D0RA05326K
23	MELONI D , FRANZONI G , OGGIANO A . Cell lines for the development of African swine fever virus vaccine candidates: an update[J]. Vaccines, 2022, 10 (5): 707. doi: 10.3390/vaccines10050707
24	TATOYAN M R , IZMAILYAN R A , SEMERJYAN A B , et al. Patterns of alveolar macrophage activation upon attenuated and virulent African swine fever viruses in vitro[J]. Comp Immunol Microbiol Infect Dis, 2020, 72, 101513. doi: 10.1016/j.cimid.2020.101513
25	徐晓曦, 范传园. 中医药对非洲猪瘟防控策略探析[J]. 中兽医学杂志, 2019, (4): 86- 89.
	XU X X , FAN C Y . Analysis on prevention and control strategy of African swine fever by traditional Chinese medicine[J]. Journal of Chinese Veterinary Medicine, 2019, (4): 86- 89.
26	李飞翔. 运用中医温病学说防控非洲猪瘟[J]. 畜牧兽医科学, 2019, (20): 82- 83.
	LI F X . Prevention and control of African swine fever with the theory of febrile disease in traditional Chinese medicine[J]. Animal Husbandry and Veterinary Science, 2019, (20): 82- 83.
27	南京中医药大学《新编温病学》编委会. 新编温病学[M]. 北京: 学苑出版社, 2003.
	Nanjing University of Traditional Chinese Medicine, "New edition of Febrile disease" Edilorial Board" . New edition of febrile diseases[M]. Beijing: Academy Press, 2003.
28	陈利平, 王发渭, 郝爱真, 等. 中医对传染性疾病的认识与防治[J]. 中华医院感染学杂志, 2008, 18 (11): 1587- 1588. doi: 10.3321/j.issn:1005-4529.2008.11.033
	CHEN L P , WANG F W , HAO A Z , et al. View of TCM on epidemic diseases and their preyention and treatment[J]. Chinese Journal of Nosocomiology, 2008, 18 (11): 1587- 1588. doi: 10.3321/j.issn:1005-4529.2008.11.033
29	王志旺, 刘雪枫, 妥海燕, 等. 育阴软肝中药对实验性肝损伤的保护作用[J]. 中国应用生理学杂志, 2015, 31 (1): 76- 79.
	WANG Z W , LIU X F , TUO H Y , et al. The protective effect of Yuyin Ruangan Decoction on experimental hepatic injury[J]. Chinese Journal of Applied Physiology, 2015, 31 (1): 76- 79.
30	王志旺, 付晓艳, 程小丽, 等. 育阴软肝颗粒剂对肝纤维化大鼠TGF-β1表达的影响[J]. 中国应用生理学杂志, 2018, 34 (2): 169- 172.
	WANG Z W , FU X Y , CHENG X L , et al. Effect of Yuyin Ruangan Granule on TGF-β1 expression in hepatic fibrosis rats[J]. Chinese Journal of Applied Physiology, 2018, 34 (2): 169- 172.
31	WEI S J , HE Q M , ZHANG Q , et al. Traditional Chinese medicine is a useful and promising alternative strategy for treatment of Sjogren's syndrome: A review[J]. J Integr Med, 2021, 19 (3): 191- 202. doi: 10.1016/j.joim.2021.01.008
32	AFONSO C L , ALCARAZ C , BRUN A , et al. Characterization of p30, a highly antigenic membrane and secreted protein of African swine fever virus[J]. Virology, 1992, 189 (1): 368- 373. doi: 10.1016/0042-6822(92)90718-5
33	SÁNCHEZ E G , PÉREZ-NÚÑEZ D , REVILLA Y . Development of vaccines against African swine fever virus[J]. Virus Res, 2019, 265, 150- 155.
34	MAZUR-PANASIUK N , WOŹNIAKOWSKI G , NIEMCZUK K . The first complete genomic sequences of African swine fever virus isolated in Poland[J]. Sci Rep, 2019, 9 (1): 4556.
35	LIU Q , MA B T , QIAN N C , et al. Structure of the African swine fever virus major capsid protein p72[J]. Cell Res, 2019, 29 (11): 953- 955.
36	SCHNEIDER W M , CHEVILLOTTE M D , RICE C M . Interferon-stimulated genes: a complex web of host defenses[J]. Annu Rev Immunol, 2014, 32, 513- 545.
37	BRASS A L , HUANG I C , BENITA Y , et al. The IFITM proteins mediate cellular resistance to influenza A H1N1 virus, West Nile virus, and dengue virus[J]. Cell, 2009, 139 (7): 1243- 1254.
38	LEWIN A R , REID L E , MCMAHON M , et al. Molecular analysis of a human interferon-inducible gene family[J]. Eur J Biochem, 1991, 199 (2): 417- 423.
39	HUANG I C , BAILEY C C , WEYER J L , et al. Distinct patterns of IFITM-mediated restriction of filoviruses, SARS coronavirus, and influenza A virus[J]. PLoS Pathog, 2011, 7 (1): e1001258.
40	ZHOU W , WHITELEY A T , DE OLIVEIRA MANN C C , et al. Structure of the human cGAS-DNA complex reveals enhanced control of immune surveillance[J]. Cell, 2018, 174 (2): 300- 311. e11.

试验用药 Experimental drug	组方 Composition	剂量/g Dosage	作用功效 Function and efficacy
连翘 Forsythia	单药	20	清热，解毒，散结，消肿
马勃Sphaerium	单药	20	清肺利咽，解毒，止血
改良育阴方(MYY)(复方1) Modified Yuyin decoction (Compound 1)	石膏	30	清热泻火
	知母	18	和石膏同用，可增强石膏的清热泻火作用
	玄参	18	清热凉血，滋阴降火，除烦，解毒
	蒲公英	18	清热解毒，利尿散结
	金银花	20	清热，解毒
	连翘	20	清热，解毒，散结，消肿
	桑叶	16	祛风清热，凉血明目
	生地黄	18	滋阴清热，凉血补血
	牡丹皮	18	清热凉血，和血消瘀
	大青叶	18	清热解毒，凉血止血
	紫苏叶	16	发散表寒，开宣肺气
	甘草	10	和中缓急，润肺，解毒，调和诸药
复方鱼腥草(复方2) Compound Houttuynia (Compound 2)	鱼腥草	16	清热解毒，利尿消肿
	黄芩	18	泻实火，除湿热，止血，安胎
	板蓝根	18	清热解毒，凉血
	连翘	20	清热，解毒，散结，消肿
	金银花	20	清热，解毒
银翘马勃散(复方3) Yinqiao Mabo powder (Compound 3)	连翘	30	清热，解毒，散结，消肿
	牛蒡子	18	疏散风热，宣肺透疹，消肿解毒
	金银花	15	清热，解毒
	射干	9	降火，解毒，散血，消痰
	马勃	6	清肺利咽，解毒，止血

目标 Target	方向 Orientation	序列(5′- 3′) Sequence
ASFV-p72	正向 Forward	ACGGCGCCCTCTAAAGGT
ASFV-p72	反向 Reverse	CATGGTCAGCTTCAAACGTTTC
Cyclophilin A	正向 Forward	AGACAAGGTTCCAAAGACAGCAG
Cyclophilin A	反向 Reverse	AGACTGAGTGGTTGGATGGCA

化学式 Formula	名称 Name	质量 Exact_mass	测量误差(ppm) Measurement error(ppm)	CAS	Library	化学结构 Structure
C₆H₄O₂	1, 2-Benzoquinone	108.021 1	2.971 703 44	583-63-1	MetaDNA
C₅H₁₀N₂O	L-Prolinamide	114.079 3	2.815 271 87	7531-52-4	MetaDNA
C₁₅H₁₈O₈	1-O-(4-coumaroyl)-β-D-glucose	326.100 2	14.021 680 47	7139-64-2	MetaDNA
C₉H₁₈N₄O₄	D-Octopine	246.132 8	29.529 685 51	34522-32-2	MetaDNA
C₁₁H₂₀N₄O₆	Nopaline	304.138 3	24.608 987 88	22350-70-5	MetaDNA
C₉H₁₃N₃O₄	Deoxycytidine	227.090 6	6.605 293 218	951-77-9	MetaDNA
C₁₉H₂₆O₂	androst-5-ene-3, 17-dione	286.193 3	23.064 020 6	571-36-8	MetaDNA
C₁₉H₂₈O₂	Androstanedione	288.208 9	1.645 827 722	846-46-8	MetaDNA
C₂₁H₂₈O₄	11-Dehydrocorticosterone	344.198 8	22.415 016 9	72-23-1	MetaDNA
C₂₇H₄₆O₄S	Cholesterol sulfate	466.311 7	7.934 606 831	1256-86-6	MetaDNA

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