植物乳杆菌HB13-2抑制白假丝酵母菌作用及机制

Inhibitory Effect and Mechanism of Lactiplantibacillus plantarum HB13-2 on Candida albicans

探究植物乳杆菌HB13-2上清液对白假丝酵母菌的抑制作用及机制。首先,通过二倍稀释法测定了上清液对白假丝酵母菌的最小抑菌浓度。继而,通过荧光增白剂对细胞壁染色观察,结果表明上清液处理使荧光强度增强,细胞壁出现损伤。通过流式细胞术检测结合荧光显微镜观察,结果显示植物乳杆菌HB13-2上清液改变了白假丝酵母菌细胞膜通透性;利用DiSC_(3)(5)检测跨膜电势,结果显示荧光强度增强,表明上清液导致跨膜电势消散。通过扫描电镜观察其微观结构,发现植物乳杆菌HB13-2上清液使菌体发生形变并伴随内容物的泄漏。采用2’,7’-二氯二氢荧光素二乙酸酯以及罗丹明-123荧光染色检测活性氧积累和线粒体膜电位,结果显示上清液使细胞内活性氧大量积累,线粒体膜电位升高。综上,植物乳杆菌HB13-2上清液可通过破坏细胞壁和细胞膜使细胞发生形变,并导致线粒体损伤,从而对白假丝酵母菌发挥抑制作用。本研究可为植物乳杆菌HB13-2作为口腔益生菌开发提供科学依据。

This study explored the inhibitory effect and mechanism of the culture supernatant of Lactiplantibacillus plantarum HB13-2 on Candida albicans.The minimum inhibitory concentration was determined by the double dilution method.Then,thefluorescent dye calcofluor white(CFW)was used to stain the cell wall and observe it.The results showed that the supernatant enhanced the fluorescence intensity and damaged the cell wall.Flow cytometry and fluorescence microscopy showed that the supernatant changed the membrane permeability of C.albicans.The transmembrane potential was detected using the fluorescent probe DiSC_(3)(5),and it was found that the fluorescence intensity was enhanced,indicating that the supernatant caused dissipation of the transmembrane potential.Through microstructural observation by scanning electron microscopy(SEM),it was found that the supernatant of Lactobacillus plantarum HB13-2 caused cellular deformation and leakage of intracellular contents.As detected byfluorescence staining with 2’,7’-dichlorodihydrofluorescin diacetate(DCFH-DA)and Rhodamine-123,the supernatant resulted in accumulation of a large amount of intracellular reactive oxygen species(ROS)and increased mitochondrial membrane potential.In conclusion,the supernatant of L.plantarum HB13-2 can deform cells by destroying the cell wall and membrane and lead to mitochondrial damage,thereby inhibiting C.albicans.This study will provide a scientific basis for the development of L.plantarum HB13-2 as an oral probiotic.