摘要
为了解由香芹酚、肉桂醛和百里香酚配制的复合植物精油对猪源肠道致病菌的体外抑菌活性及其对细胞炎症反应的影响,分别采用纸片扩散法、宏量肉汤稀释法测定复合植物精油对3种致病菌的抑菌圈直径和最小抑菌浓度(MIC),并采用透射电镜观察复合植物精油作用后3种致病菌形态结构变化,将3种致病菌与猪小肠上皮细胞共孵育建立体外炎症损伤模型,通过荧光定量PCR检测复合植物精油对该模型炎性细胞因子mRNA表达的影响。结果表明,复合植物精油对大肠埃希氏菌、金黄色葡萄球菌、沙门氏菌的抑菌圈直径分别为(15.60±0.79)mm、(19.17±0.85)mm、(11.43±0.31)mm,其对3种致病菌的最小抑菌浓度均为0.42μL/mL;经复合植物精油处理6 h后,3种细菌细胞壁、膜结构完整性被破坏,细胞内容物泄漏,细胞产生空腔甚至空化;与对照组相比,炎症损伤模型中细胞IL-1β、IL-6、TNF-α的mRNA水平极显著上调(P<0.01),与复合植物精油共孵育6 h后炎症损伤模型中细胞IL-1β、IL-6、TNF-α的mRNA水平极显著降低(P<0.01)。结果表明,复合植物精油对猪源肠道致病菌有显著抑制作用,且能显著降低致病菌感染的猪小肠上皮细胞炎性细胞因子表达水平。
The essential oil compounds consists of three monomers:carvacrol,cinnamaldehyde and thymol.The experiment was conducted to study antibacterial characteristics of essential oil compounds against porcine intestinal pathogenic bacteria in vitro and its effect of the cellular inflammatory response.The bacteriostatic circle diameter and minimum inhibitory concentration(MIC)of essential oil compounds against three pathogens were determined by the filter paper method and broth dilution method,respectively.Morphological and structural changes of the three pathogens were observed by transmission electron microscopy after treatment of essential oil compounds.Inflammation injury model was established by co-incubation of porcine pathogenic Escherichia coli,Staphylococcus aureus and Salmonella with porcine intestinal epithelial cells.The effects of compound essential oil on the mRNA expressions of inflammatory cytokines in porcine intestinal epithelial cells were detected by fluorescence quantitative PCR.Bacteriostatic circle diameters of essential oil compounds against Escherichia coli,Staphylococcus aureus and Salmonella were(15.60±0.79)mm,(19.17±0.85)mm and(11.43±0.31)mm,respectively.The MIC of essential oil compounds on three pathogens were 0.42μL/mL.Cell wall and cell membrane’s structural integrity of three pathogens were destroyed after 6h treatment with essential oil compounds.The leakage of cell content causes the production of cell cavities or even cavitation.Compared with the control group,the mRNA levels of IL-1β,IL-6 and TNF-αin the inflammatory injury model were extremely significantly up-regulated(P<0.01).And the mRNA levels of IL-1β,IL-6 and TNF-αin porcine intestinal epithelial cell inflammation were extremely significantly decreased under the co-culture with essential oil compounds(P<0.01).To sum up,the results showed that the essential oil compounds has a significant inhibitory effect on porcine pathogenic bacteria.And it can dramatically reduce the expression levels of inflammatory cytokines in porcine intestinal epithelial cells infected with pathogenic bacteria.
作者
杨阳
聂丰秋
邹盼盼
史立建
王朝霞
王春江
王迈
曹省艳
齐雪峰
YANG Yang;NIE Feng-qiu;ZOU Pan-pan;SHI Li-jian;WANG Zhao-xia;WANG Chun-jiang;WANG Mai;CAO Sheng-yan;QI Xue-feng(College of Veterinary Medicine,Northwest A&F University,Yangling,Shaanxi,712100,China;Hebei Veyong Pharmaceutical Co.,Ltd,Shijiazhuang,Hebei,052165,China;Shijiazhuang Agricultural Comprehensive Administrative Law Enforcement Detachment,Shijiazhuang,Hebei,050026,China;Mianyang Animal Disease Prevention and Control Center,Mianyang,Sichuan,621000,China)
出处
《动物医学进展》
北大核心
2022年第6期79-85,共7页
Progress In Veterinary Medicine
基金
校企合作项目(K4030220403)。
关键词
复合植物精油
抑菌活性
炎症
essential oil compound
antibacterial activity
inflammation
