Zhang Zhen, Zhao Yuanyuan, Chen Xueqin, Li Wei, Li Wen, Du Jianming, Wang Li
College of Food Science and Engineering, Gansu Agricultural University, Lanzhou 730070, China.
Foods. 2022 Jul 27;11(15):2234. doi: 10.3390/foods11152234.
Salmonella is an important pathogen causing food poisoning. Food safety and health are the themes of today′s society. As a class of food-borne pathogens, Salmonella enteritidis had become one of the common zoonotic pathogens. Cinnamon essential oil (CEO) had been reported as an antibacterial agent, but there are few studies on its antibacterial mechanism. This study investigated the effects of CEO on oxidative damage and outer membrane protein genes of Salmonella enteritidis cells. First, the reactive oxygen species content in bacteria treated with different concentrations of cinnamon essential oil was determined by fluorescence spectrophotometry, and the effects of superoxide dismutase (SOD), catalase (CAT) and superoxide dismutase (SOD), and catalase (CAT) and peroxidase (POD) were determined by the kit method. The activity of POD and the content of malondialdehyde (MDA) were investigated to investigate the oxidative damage of CEO to Salmonella enteritidis cells. By analyzing the effect of CEO on the Salmonella enteritidis cell membrane’s outer membrane protein gene expression, the mechanism of CEO′s action on the Salmonella enteritidis cell membrane was preliminarily discussed. The results showed that CEO treatment had an obvious oxidative damaging effect on Salmonella enteritidis. Compared with the control group, the increase in CEO concentration caused a significant increase in the bacteria ROS content. The observation technique experiment found that with the increase in CEO concentration, the number of stained cells increased, which indicated that CEO treatment would increase the ROS level in the cells, and it would also increase with the increase in CEO concentration, thus causing the oxidation of cells and damage. In addition, CEO treatment also caused the disruption of the balance of the cellular antioxidant enzymes (SOD, CAT, POD) system, resulting in an increase in the content of MDA, a membrane lipid metabolite, and increased protein carbonylation, which ultimately inhibited the growth of Salmonella enteritidis. The measurement results of cell membrane protein gene expression levels showed that the Omp genes to be detected in Salmonella enteritidis were all positive, which indicated that Salmonella enteritidis carried these four genes. Compared with the control group, the relative expressions of OmpF, OmpA and OmpX in the CEO treatment group were significantly increased (p < 0.05), which proved that the cell function was disturbed. Therefore, the toxicity of CEO to Salmonella enteritidis could be attributed to the damage of the cell membrane and the induction of oxidative stress at the same time. It was speculated that the antibacterial mechanism of CEO was the result of multiple effects. This work was expected to provide a theoretical basis for the development of new natural food preservatives and the prevention and control of Salmonella enteritidis.
沙门氏菌是引起食物中毒的重要病原体。食品安全与健康是当今社会的主题。作为一类食源性病原体,肠炎沙门氏菌已成为常见的人畜共患病原体之一。肉桂精油(CEO)已被报道为一种抗菌剂,但其抗菌机制的研究较少。本研究探讨了肉桂精油对肠炎沙门氏菌细胞氧化损伤及外膜蛋白基因的影响。首先,采用荧光分光光度法测定不同浓度肉桂精油处理后细菌内活性氧的含量,并用试剂盒法测定超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、过氧化物酶(POD)的活性及丙二醛(MDA)的含量,以研究肉桂精油对肠炎沙门氏菌细胞的氧化损伤。通过分析肉桂精油对肠炎沙门氏菌细胞膜外膜蛋白基因表达的影响,初步探讨了肉桂精油作用于肠炎沙门氏菌细胞膜的机制。结果表明,肉桂精油处理对肠炎沙门氏菌有明显的氧化损伤作用。与对照组相比,随着肉桂精油浓度的增加,细菌活性氧含量显著增加。观察技术实验发现,随着肉桂精油浓度的增加,染色细胞数量增多,这表明肉桂精油处理会使细胞内活性氧水平升高,且随肉桂精油浓度增加而升高,从而导致细胞氧化损伤。此外,肉桂精油处理还导致细胞抗氧化酶(SOD、CAT、POD)系统平衡破坏,导致膜脂代谢产物MDA含量增加,蛋白质羰基化增加,最终抑制了肠炎沙门氏菌的生长。细胞膜蛋白基因表达水平测定结果显示,肠炎沙门氏菌中检测到的Omp基因均为阳性,表明肠炎沙门氏菌携带这四个基因。与对照组相比,肉桂精油处理组中OmpF、OmpA和OmpX的相对表达量显著增加(p < 0.05),证明细胞功能受到干扰。因此,肉桂精油对肠炎沙门氏菌的毒性可归因于对细胞膜的损伤和同时诱导的氧化应激。推测肉桂精油的抗菌机制是多种作用的结果。本研究有望为新型天然食品防腐剂的开发及肠炎沙门氏菌的防控提供理论依据。