甘草查尔酮A对副猪嗜血杆菌的体内外抑制作用
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摘要
探讨甘草查尔酮A对副猪嗜血杆菌体内外生长的影响。将副猪嗜血杆菌于分别含10、20、50、100、200μg/mL甘草查尔酮A的TSA液体培养基中培养,采用分光光度法分析副猪嗜血杆菌生长和生物被膜形成的变化;以1、10、100 mg/kg甘草查尔酮A经口灌胃小鼠,分别在3 d、5 d和7 d称重,并计算脏器指数,Hiseq高通量测序技术分析肠道菌群等生长性能,Bliss法测定副猪嗜血杆菌对小鼠半数致死量(LD_(50));检测甘草查尔酮A对小鼠清除0.01×LD_(50)副猪嗜血杆菌感染和抵抗100×LD_(50)副猪嗜血杆菌感染的影响。结果表明,各浓度甘草查尔酮A对副猪嗜血杆菌的生长速率和生物被膜形成的影响差异均无显著差异。所有剂量组小鼠给予甘草查尔酮A处理后,一般临床症状、体重和脏器指数无统计学意义。灌喂甘草查尔酮A有增加小鼠肠道菌群丰富度的趋势,但降低拟杆菌门丰度而增加厚壁菌门丰度。小鼠清除0.01×LD_(50)副猪嗜血杆菌感染的能力呈剂量依赖性,而100×LD_(50)的副猪嗜血杆菌感染小鼠后随着甘草查尔酮A浓度增加可呈剂量依赖延迟小鼠死亡时间。表明甘草查尔酮A对副猪嗜血杆菌体外生长无显著的抑制作用,同时对小鼠生长性能亦无明显影响,但可增加肠道菌群丰富度而提高小鼠抵抗副猪嗜血杆菌感染的能力。
To investigate the effect of licochalcone A on growth performance and inhibition of Haemophilus parasuis in vitro and in vivo,after coculture with Licochalcone A(10 μg/mL,20 μg/mL,50 μg/mL,100 μg/mL,and 200 μg/mL),the growth velocity and biofilm formation of Haemophilus parasuis were measured by spectrophotometric method.Body weight,viscera index,and intestinal microbiota in mice given with licochalcone A at low(1 mg/kg), middle(10 mg/kg) and high doses(100 mg/kg) were determined by weighing method and Hiseq sequencing.The median lethal dosage(LD_(50)) was calculated by Bliss method.The bacterial clearance for 0.01×LD_(50) Haemophilus parasuis and protective efficacy against 100×LD_(50) Haemophilus parasuis were evaluated in mice given with licochalcone A.The difference was not statistically significant in the effect of licochalcone A on the growth velocity and biofilm formation of Haemophilus parasuis H12 L1.The clinical condition of disease was no change in the treatment group.Body weight and viscera indexes(liver,lung,heart,kidney and thymus) in mice were increased slightly(P>0.05).Oral administration of licochalcone A increased the diversity of intestinal microbiota in mice with the decrease of Bacteroidetes and the increase of Firmicutes.LD_(50) for intraperitoneal inoculation of Haemophilus parasuis was 8.72×10~7 CFU/mL.The bacterial clearance of mice infected with 0.01×LD_(50) Haemophilus parasuis was gradually strengthened.And the increase of mean survival time was found in mice infected with 100×LD_(50) Haemophilus parasuis after administration of licochalcone A.The results showed that licochalcone A had low inhibitory effect on the growth of Haemophilus parasuis in vitro and in vivo and no significant effect on growth performance of mice.However,Licochalcone A slightly increased the diversity of intestinal microbiota against infection of Haemophilus parasuis in mice.
To investigate the effect of licochalcone A on growth performance and inhibition of Haemophilus parasuis in vitro and in vivo,after coculture with Licochalcone A(10 μg/mL,20 μg/mL,50 μg/mL,100 μg/mL,and 200 μg/mL),the growth velocity and biofilm formation of Haemophilus parasuis were measured by spectrophotometric method.Body weight,viscera index,and intestinal microbiota in mice given with licochalcone A at low(1 mg/kg), middle(10 mg/kg) and high doses(100 mg/kg) were determined by weighing method and Hiseq sequencing.The median lethal dosage(LD_(50)) was calculated by Bliss method.The bacterial clearance for 0.01×LD_(50) Haemophilus parasuis and protective efficacy against 100×LD_(50) Haemophilus parasuis were evaluated in mice given with licochalcone A.The difference was not statistically significant in the effect of licochalcone A on the growth velocity and biofilm formation of Haemophilus parasuis H12 L1.The clinical condition of disease was no change in the treatment group.Body weight and viscera indexes(liver,lung,heart,kidney and thymus) in mice were increased slightly(P>0.05).Oral administration of licochalcone A increased the diversity of intestinal microbiota in mice with the decrease of Bacteroidetes and the increase of Firmicutes.LD_(50) for intraperitoneal inoculation of Haemophilus parasuis was 8.72×10~7 CFU/mL.The bacterial clearance of mice infected with 0.01×LD_(50) Haemophilus parasuis was gradually strengthened.And the increase of mean survival time was found in mice infected with 100×LD_(50) Haemophilus parasuis after administration of licochalcone A.The results showed that licochalcone A had low inhibitory effect on the growth of Haemophilus parasuis in vitro and in vivo and no significant effect on growth performance of mice.However,Licochalcone A slightly increased the diversity of intestinal microbiota against infection of Haemophilus parasuis in mice.
引文
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[2] Kuehn J S,Register K B,Phillips G J.Draft genome sequences for 10 isolates of the swine pathogen Haemophilus parasuis[J].Genome Announc,2013,1(5pii):e00739-13.
[3] Wang L,Yang R,Yuan B,et al.The antiviral and antimicrobial activities of licorice,a widely-used Chinese herb[J].Acta Pharm Sin B,2015,5(4):10-315.
[4] Bello-ortí B,Deslandes V,Tremblay Y D,et al.Biofilm formation by virulent and non-virulent strains of Haemophilus parasuis[J].Vet Res,2014,45(104):1-14.
[5] Chen X,Liu Z,Meng R,et al.Antioxidative and anticancer properties of licochalcone A from licorice[J].J Ethnopharmacol,2017,198:331-337.
[6] Si H,Xu C,Zhang J,et al.Licochalcone A:An effective and low-toxicity compound against Toxoplasma gondii in vitro and in vivo[J].Int J Parasitol Drugs Drug Resist,2018,8(2):238-245.
[7] Seleem D,Benso B,Noguti J,et al.In vitro and in vivo antifungal activity of lichochalcone-A against Candida albicans biofilms[J].PLoS One,2016,11(6):e0157188.
[8] 申凤鸽.甘草查耳酮A抗金黄色葡萄球菌生物被膜的分子机制初步研究[D].吉林长春:吉林大学,2013.
[9] Tsukiyama R,Katsura H,Tokuriki N,et al.Antibacterial activity of licochalcone A against spore-forming bacteria[J].Antimicrob Agents Ch,2002,46(5):1226-1230.