Reyes Alisha Wehdnesday Bernardo, Simborio Hannah Leah Tadeja, Hop Huynh Tan, Arayan Lauren Togonon, Min Won Gi, Lee Hu Jang, Rhee Man Hee, Chang Hong Hee, Kim Suk
Institute of Animal Medicine, College of Veterinary Medicine, Gyeongsang National University, Jinju 52828, Korea.
Department of Veterinary Paraclinical Sciences, College of Veterinary Medicine, University of the Philippines Los Baños, Los Baños 4031, Philippines.
J Vet Sci. 2016 Sep 30;17(3):315-21. doi: 10.4142/jvs.2016.17.3.315.
Korean red ginseng (KRG) has long been used in traditional Korean and Oriental medicine. However, the anti-bacterial mechanism and therapeutic efficiency of KGR for intracellular Brucella infection are still unclear. In this study, the bactericidal activity of Korean red ginseng acidic polysaccharide (RGAP) on Brucella (B.) abortus and its cytotoxic effects on RAW 264.7 cells were evaluated. In addition, B. abortus internalization and intracellular replication in macrophages were investigated after RGAP treatment. RGAP-incubated cells displayed a marked reduction in the adherence, internalization and intracellular growth of B. abortus in macrophages. Furthermore, decreased F-actin fluorescence was observed relative to untreated B. abortus-infected cells. Western blot analysis of intracellular signaling proteins revealed reduced ERK, JNK and p38α phosphorylation levels in B. abortus-infected RGAP-treated cells compared to the control. Moreover, elevated co-localization of B. abortus-containing phagosomes with lysosome-associated membrane protein 1 (LAMP-1) were observed in RGAP-treated cells compared with the control. Overall, the results of this study suggest that RGAP can disrupt phagocytic activity of B. abortus via suppression of mitogen-activated protein kinases (MAPKs) signaling proteins ERK, JNK and p38 levels and inhibit intracellular replication of B. abortus by enhancing phagolysosome fusion, which may provide an alternative control of brucellosis.
韩国红参(KRG)长期以来一直应用于传统的韩国和东方医学。然而,韩国红参对细胞内布鲁氏菌感染的抗菌机制和治疗效果仍不清楚。在本研究中,评估了韩国红参酸性多糖(RGAP)对流产布鲁氏菌的杀菌活性及其对RAW 264.7细胞的细胞毒性作用。此外,还研究了RGAP处理后巨噬细胞中流产布鲁氏菌的内化和细胞内复制情况。经RGAP孵育的细胞中,流产布鲁氏菌在巨噬细胞中的黏附、内化和细胞内生长均显著降低。此外,与未处理的流产布鲁氏菌感染细胞相比,观察到F-肌动蛋白荧光减弱。细胞内信号蛋白的蛋白质印迹分析显示,与对照组相比,经RGAP处理的流产布鲁氏菌感染细胞中ERK、JNK和p38α的磷酸化水平降低。此外,与对照组相比,在经RGAP处理的细胞中观察到含流产布鲁氏菌的吞噬体与溶酶体相关膜蛋白1(LAMP-1)的共定位增加。总体而言,本研究结果表明,RGAP可通过抑制丝裂原活化蛋白激酶(MAPKs)信号蛋白ERK、JNK和p38的水平来破坏流产布鲁氏菌的吞噬活性,并通过增强吞噬溶酶体融合来抑制流产布鲁氏菌的细胞内复制,这可能为布鲁氏菌病提供一种替代控制方法。