Verbascoside lowers Streptococcus suis serotype 2 pathogenicity in mice by inhibiting hemolytic activity of suilysin

doi:10.3969/j.issn.1004-1524.2022.08.05

Acta Agriculturae Zhejiangensis ›› 2022, Vol. 34 ›› Issue (8): 1609-1616.DOI: 10.3969/j.issn.1004-1524.2022.08.05

• Animal Science • Previous Articles Next Articles

Verbascoside lowers Streptococcus suis serotype 2 pathogenicity in mice by inhibiting hemolytic activity of suilysin

ZHAN Jiafei¹(), XU Kui², ZHANG Lei¹, XIA Jieying¹, HONG Yang¹, DONG Han¹, LIU Yanglu¹, ZHOU Jing¹, YUAN Mingming¹, WANG Yongjin¹, YAN Liangchun¹^,^*()

1. Center of Experimental Animals, Sichuan Academy of Chinese Medicine Sciences, Chengdu 610041, China
2. College of Veterinary Medicine, Sichuan Agricultural University, Chengdu 611130, China

Received:2021-12-22 Online:2022-08-25 Published:2022-08-26
Contact: YAN Liangchun

Abstract

Abstract:

The aim of this study is to explore the effect of verbascoside (VBS), a natural compound, on the pathogenicity of Streptococcus suis serotype 2 (SS2). By determining the minimum inhibitory concentration (MIC), growth curve of SS2 and hemolysis activity of erythrocytes, is was found that the low concentration of VBS significantly inhibited the hemolytic activity of SS2 without interfering with its growth. Western blotting showed that VBS had no influence on the expression of suilysin (SLY) in SS2. It was predicted by molecular docking that VBS could be embedded into the domain 4 of SLY, which was through the formation of three hydrogen bonds between VBS and two residues (SER-388, GLN-441) of SLY. And it was further confirmed that VBS directly inhibited the hemolytic activity of SLY in vitro. Moreover, VBS significantly increased the survival rate of mice infected with SS2 in vivo. In conclusion, without affecting the growth of SS2, VBS can inhibit the hemolytic activity of SLY to reduce the pathogenicity of SS2 in mice.

Key words: Streptococcus suis serotype 2, suilysin, verbascoside, pathogenicity

CLC Number:

S858.28

ZHAN Jiafei, XU Kui, ZHANG Lei, XIA Jieying, HONG Yang, DONG Han, LIU Yanglu, ZHOU Jing, YUAN Mingming, WANG Yongjin, YAN Liangchun. Verbascoside lowers Streptococcus suis serotype 2 pathogenicity in mice by inhibiting hemolytic activity of suilysin[J]. Acta Agriculturae Zhejiangensis, 2022, 34(8): 1609-1616.

Figures/Tables 7

Fig.1 Growth curves of SS2 treated with different concentrations of VBS

Fig.2 Hemolytic activity in supernatant of VBS and SS2 co-culture system NC, Negative control. * meaned significant difference compared with VBS-free group (P<0.05), ** meaned extremely significant difference compared with VBS-free group (P<0.01). The same as below.

Fig.3 Expression of SLY in supernatant of VBS and SS2 co-culture system

Fig.4 The molecular docking conformation of VBS with SLY A, 2D chemical structure of VBS; B, 3D chemical structure of VBS; C, Details of binding between the hydroxyl group of VBS and the residues of SLY.

Fig.5 Expression and purification of recombinant SLY M, Protein molecular weight marker; 1, Lysate of IPTG-induced E.coil BL21 expressing recombinant SLY; 2, Purified recombinant SLY.

Fig.6 Effect of VBS on the hemolytic activity of purified SLY

Fig.7 Effect of VBS on the survival rate of mice infected with SS2

References 34

[1]	VANIER G, SEGURA M, FRIEDL P, et al. Invasion of porcine brain microvascular endothelial cells by Streptococcus suis serotype 2[J]. Infection and Immunity, 2004, 72(3): 1441-1449. DOI URL
[2]	FITTIPALDI N, SEGURA M, GRENIER D, et al. Virulence factors involved in the pathogenesis of the infection caused by the swine pathogen and zoonotic agent Streptococcus suis[J]. Future Microbiology, 2012, 7(2): 259-279. DOI URL
[3]	GUO G L, KONG X W, DU D C, et al. Genome-wide association study identifies the virulence-associated marker in Streptococcus suis serotype 2[J]. Infection, Genetics and Evolution, 2021, 92: 104894.
[4]	GOYETTE-DESJARDINS G, AUGER J P, XU J G, et al. Streptococcus suis, an important pig pathogen and emerging zoonotic agent-an update on the worldwide distribution based on serotyping and sequence typing[J]. Emerging Microbes & Infections, 2014, 3(1): 1-20.
[5]	徐魁, 王巍, 文心田, 等. 四川主要养猪地区猪2型链球菌血清流行病学调查[J]. 中国预防兽医学报, 2017, 39(5): 370-373.
	XU K, WANG W, WEN X T, et al. Epidemiological investigation of swine Streptococcus suis type 2 in Sichuan region[J]. Chinese Journal of Preventive Veterinary Medicine, 2017, 39(5): 370-373. (in Chinese with English abstract)
[6]	MUSTHAFA K S, VORAVUTHIKUNCHAI S P. Anti-virulence potential of eugenyl acetate against pathogenic bacteria of medical importance[J]. Antonie Van Leeuwenhoek, 2015, 107(3): 703-710. DOI URL
[7]	TENENBAUM T, ASMAT T M, SEITZ M, et al. Biological activities of suilysin: role in Streptococcus suis pathogenesis[J]. Future Microbiology, 2016, 11: 941-954. DOI URL
[8]	TANABE S I, GOTTSCHALK M, GRENIER D. Hemoglobin and Streptococcus suis cell wall act in synergy to potentiate the inflammatory response of monocyte-derived macrophages[J]. Innate Immunity, 2008, 14(6): 357-363. DOI URL
[9]	XU L F, HUANG B, DU H M, et al. Crystal structure of cytotoxin protein suilysin from Streptococcus suis[J]. Protein & Cell, 2010, 1(1): 96-105.
[10]	TAKEUCHI D, AKEDA Y, NAKAYAMA T, et al. The contribution of suilysin to the pathogenesis of Streptococcus suis meningitis[J]. The Journal of Infectious Diseases, 2013, 209(10): 1509-1519. DOI URL
[11]	LAFRANCE M E, FARROW M A, CHANDRASEKARAN R, et al. Identification of an epithelial cell receptor responsible for Clostridium difficile TcdB-induced cytotoxicity[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(22): 7073-7078.
[12]	GOTTSCHALK M, SEGURA M, XU J G. Streptococcus suis infections in humans: the Chinese experience and the situation in North America[J]. Animal Health Research Reviews, 2007, 8(1): 29-45. DOI URL
[13]	PAOLA R D I, OTERI G, MAZZON E, et al. Effects of verbascoside, biotechnologically purified by Syringa vulgaris plant cell cultures, in a rodent model of periodontitis[J]. The Journal of Pharmacy and Pharmacology, 2011, 63(5): 707-717. DOI URL
[14]	袁雅婷, 胡贵丽, 丁鹏, 等. 毛蕊花糖苷的生理功能及其在动物生产中的应用[J]. 动物营养学报, 2016, 28(12): 3777-3783.
	YUAN Y T, HU G L, DING P, et al. Biological functions of verbascoside and its application in animal production[J]. Chinese Journal of Animal Nutrition, 2016, 28(12): 3777-3783. (in Chinese with English abstract)
[15]	姜醒, 鲁丽敏, 张春蕾, 等. 连翘中具有抗菌活性的苯乙醇苷类化学成分研究[J]. 中国现代中药, 2017, 19(5): 642-647.
	JIANG X, LU L M, ZHANG C L, et al. Antibacterial phenylethanoid glycosides from fruits of Forsythia suspensa[J]. Modern Chinese Medicine, 2017, 19(5): 642-647. (in Chinese with English abstract)
[16]	HUMPHRIES R M, AMBLER J, MITCHELL S L, et al. CLSI methods development and standardization working group best practices for evaluation of antimicrobial susceptibility tests[J]. Journal of Clinical Microbiology, 2018, 56(4): e01934-e01917.
[17]	ZHANG Y Y, ZONG B B, WANG X R, et al. Fisetin lowers Streptococcus suis serotype 2 pathogenicity in mice by inhibiting the hemolytic activity of suilysin[J]. Frontiers in Microbiology, 2018, 9: 1723. DOI URL
[18]	ZHAO X R, LI H E, WANG J F, et al. Verbascoside alleviates pneumococcal pneumonia by reducing pneumolysin oligomers[J]. Molecular Pharmacology, 2016, 89(3): 376-387. DOI URL
[19]	RASKO D A, SPERANDIO V. Anti-virulence strategies to combat bacteria-mediated disease[J]. Nature Reviews Drug Discovery, 2010, 9 (2): 117-128. DOI URL
[20]	HUANG J H, SHANG K X, KASHIF J, et al. Genetic diversity of Streptococcus suis isolated from three pig farms of China obtained by acquiring antibiotic resistance genes[J]. Journal of the Science of Food and Agriculture, 2015, 95(7): 1454-1460. DOI URL
[21]	HU P, YANG M, ZHANG A D, et al. Comparative genomics study of multi-drug-resistance mechanisms in the antibiotic-resistant Streptococcus suis R61 strain[J]. PLoS One, 2011, 6(9): e24988.
[22]	BONIFAIT L, GRIGNON L, GRENIER D. Fibrinogen induces biofilm formation by Streptococcus suis and enhances its antibiotic resistance[J]. Applied and Environmental Microbiology, 2008, 74(15): 4969-4972. DOI URL
[23]	CEGELSKI L, MARSHALL G R, ELDRIDGE G R, et al. The biology and future prospects of antivirulence therapies[J]. Nature Reviews Microbiology, 2008, 6 (1): 17-27. DOI URL
[24]	苏永南. 中医药治疗传染病的现状分析与对策研究: 基于福州市调查[D]. 福州: 福建医科大学, 2011.
	SU Y N. Research on the status quo and optimization strategies of treatment of infectious diseases with TCM: based on survey data in Fuzhou[D]. Fuzhou: Fujian Medical University, 2011. (in Chinese with English abstract)
[25]	张煜卓, 戚涵姝, 谷笑雨, 等. 分子对接在药物虚拟筛选中的应用进展[J]. 广州化学, 2017, 42(6): 62-67.
	ZHANG Y Z, QI H S, GU X Y, et al. Progress of molecular docking application in drug virtual screening[J]. Guangzhou Chemistry, 2017, 42(6): 62-67. (in Chinese with English abstract)
[26]	LI G, LU G J, QI Z M, et al. Morin attenuates Streptococcus suis pathogenicity in mice by neutralizing suilysin activity[J]. Frontiers in Microbiology, 2017, 8: 460.
[27]	LI G, SHEN X, WEI Y H, et al. Quercetin reduces Streptococcus suis virulence by inhibiting suilysin activity and inflammation[J]. International Immunopharmacology, 2019, 69: 71-78. DOI URL
[28]	LI G, WANG G Z, SI X S, et al. Inhibition of suilysin activity and inflammation by myricetin attenuates Streptococcus suis virulence[J]. Life Sciences, 2019, 223: 62-68. DOI URL
[29]	WANG G Z, LIU H M, LIU Y, et al. Formononetin alleviates Streptococcus suis infection by targeting suilysin[J]. Microbial Pathogenesis, 2020, 147: 104388.
[30]	WANG G Z, GAO Y W, WU X H, et al. Inhibitory effect of piceatannol on Streptococcus suis infection both in vitro and in vivo[J]. Frontiers in Microbiology, 2020, 11: 593588.
[31]	LIU Y, WANG H, GAO J, et al. Cryptotanshinone ameliorates the pathogenicity of Streptococcus suis by targeting suilysin and inflammation[J]. Journal of Applied Microbiology, 2021, 130(3): 736-744. DOI URL
[32]	林玲, 李春玲, 臧莹安, 等. 猪链球菌毒力因子研究进展[J]. 动物医学进展, 2020, 41(9): 82-86.
	LIN L, LI C L, ZANG Y G, et al. Progress on virulence factors of Streptococcus suis[J]. Progress in Veterinary Medicine, 2020, 41(9): 82-86. (in Chinese with English abstract)
[33]	TWETEN R K. Cholesterol-dependent cytolysins, a family of versatile pore-forming toxins[J]. Infection and Immunity, 2005, 73(10): 6199-6209. DOI URL
[34]	TWETEN R K, PARKER M W, JOHNSON A E. The cholesterol-dependent cytolysins[J]. Current Topics in Microbiology and Immunology, 2001, 257(13): 15-33.

Verbascoside lowers Streptococcus suis serotype 2 pathogenicity in mice by inhibiting hemolytic activity of suilysin

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