苦参碱对牛源无乳链球菌的体外抑菌活性分析

doi:10.11843/j.issn.0366-6964.2023.03.040

摘要/Abstract

摘要： 旨在探讨苦参碱对牛源无乳链球菌的体外抑菌活性及其对无乳链球菌毒力因子的影响，为中药单体抑菌产品的开发与应用提供理论依据。通过测定最小抑菌浓度和抑菌曲线分析苦参碱对无乳链球菌的抑菌活性；同时利用全血杀伤试验、黏附试验、生物膜染色和透射电镜观察等方法，探究苦参碱对无乳链球菌抵御机体杀伤、对宿主的黏附和生物膜形成的影响。结果显示，苦参碱在体外能够有效地抑制无乳链球菌(ATCC13813)的增殖，其对无乳链球菌的最小抑菌浓度为4 mg·mL^-1，苦参碱可显著降低无乳链球菌生物膜的形成；透射电镜可见无乳链球菌的细胞膜表面皱缩，胞壁光滑变薄，生物膜的形成受到明显抑制。苦参碱可显著抑制无乳链球菌对于乳腺上皮细胞的黏附。荧光定量 PCR 结果表明，苦参碱可显著降低与侵袭和免疫逃避相关毒力基因ScpB、CpsA、CylE和CAMP的表达(P<0.05)，显著上调CpsE基因的表达(P<0.05)。此外，苦参碱可显著降低与毒力因子黏附作用相关的Bac、Bca、Lmb、BIBA、PI-2b和FbsB基因的表达(P<0.05)。以上结果表明，苦参碱在体外能够有效抑制无乳链球菌(ATCC13813)的增殖，破坏细胞的完整性，降低菌株的侵袭力，从而达到抗菌的效果。

关键词: 苦参碱, 无乳链球菌, 全血杀伤, 黏附能力

Abstract: This study aimed to investigate the antibacterial activity of matrine against bovine Streptococcus agalactiae in vitro, and to provide theoretical basis for the development of monomer antibacterial products of Chinese herbal medicine. In this study, the bacteriostatic activity of matrine against Streptococcus agalactis was analyzed by determining the minimum inhibitory concentration and bacteriostatic curve. In the mean time, whole blood killing test, adhesion test, biofilm staining and electron microscopic observation were used to explore the effects of matrine on the resistance of Streptococcus agalactis to organism killing, host adhesion and biofilm formation. The results showed that matrine could effectively inhibit the proliferation of Streptococcus agalactiae (ATCC13813) in vitro, and the minimum inhibitory concentration of matrine against Streptococcus agalactiae was 4 mg·mL^-1, the biofilm formation of S. agalactiae was decreased as well. Scanning transmission electron microscopy showed that the surface of the cell membrane of Streptococcus agalactiae shrank, the cell wall became smooth and thin, and the formation of capsule was significantly inhibited. Matrine could significantly inhibit the adhesion of Streptococcus agalactiae to mammary epithelial cells. Quantitative PCR showed that matrine significantly decreased the expression of virulence genes ScpB, CpsA, CylE and CAMP which related to invasion and immune evasion (P<0.05), and significantly up-regulated the expression of CpsE gene (P<0.05). In addition, matrine significantly decreased the expression of Bac, Bca, Lmb, BIBA, PI-2b and FbsB genes which related to the adhesion of virulence factors (P<0.05). These results indicate that matrine can achieve the antibacterial effect by inhibiting the proliferation of Streptococcus agalactiae (ATCC13813) in vitro, destroying the integrity thus decreasing the invasion of the strain.

Key words: matrine, Streptococcus agalactiae, whole blood killing, adhesion ability

中图分类号:

S853.74

霍相羽, 高家瑞, 靳佳祺, 李燕, 高原, 刘梦涵, 蒋林树, 童津津. 苦参碱对牛源无乳链球菌的体外抑菌活性分析[J]. 畜牧兽医学报, 2023, 54(3): 1300-1309.

HUO Xiangyu, GAO Jiarui, JIN Jiaqi, LI Yan, GAO Yuan, LIU Menghan, JIANG Linshu, TONG Jinjin. Antibacterial Activity of Martine against Bovine Streptococcus agalactiae in vitro[J]. Acta Veterinaria et Zootechnica Sinica, 2023, 54(3): 1300-1309.

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