Jang Kyung-Jun, Choi Sang Hoon, Yu Gyeong Jin, Hong Su Hyun, Chung Yoon Ho, Kim Cheol-Hong, Yoon Hyun-Min, Kim Gi-Young, Kim Byung Woo, Choi Yung Hyun
Department of Acupuncture and Moxibustion, Dongeui University College of Korean Medicine, Busan 614-052, Republic of Korea.
Department of Biochemistry, Dongeui University College of Korean Medicine, Busan 614-052, Republic of Korea.
Exp Ther Med. 2016 Mar;11(3):1109-1115. doi: 10.3892/etm.2015.2965. Epub 2015 Dec 29.
Ginseng, the root of C.A. Meyer (Araliaceae), is a widely known traditional medicine that has been utilized throughout Asia for several thousand years. Ginseng saponins exert various important pharmacological effects regarding the control of a number of diseases. The aim of the present study was to identify the anti-inflammatory effects of total saponins extracted from ginseng (TSG) on lipopolysaccharide (LPS)-stimulated mouse RAW 264.7 macrophages. The inhibitory effects of TSG on LPS-induced nitric oxide (NO) production and LPS-induced tumor necrosis factor-α (TNF-α) and interleukin-1β (IL-1β) protein expression were determined by measuring the levels of nitrite and enzyme-linked immunosorbent assays, respectively. Furthermore, the effects of TSG on the mRNA expression levels and localizations of inducible NO synthase (iNOS), IL-1β and TNF-α, and their upstream signaling proteins, including nuclear factor-κB (NF-κB) and mitogen-activated protein kinases (MAPKs), were investigated by reverse transcription-polymerase chain reaction and western blotting, respectively. Following stimulation with LPS, elevated levels of NO production were detected in RAW 264.7 cells; however, TSG pretreatment significantly inhibited the production of NO (P<0.05), by suppressing the expression of iNOS. In addition, LPS-stimulated TNF-α and IL-1β production was significantly reduced by TSG (P<0.05). In the LPS-stimulated RAW 264.7 cells, NF-κB was translocated from the cytosol to the nucleus, whilst TSG pretreatment induced the sequestration of NF-κB in the cytosol by inhibiting inhibitor of κB degradation. TSG also contributed to downregulation of MAPKs in LPS-stimulated RAW 264.7 cells. These results suggested that TSG may exert anti-inflammatory activity, and that TSG may be considered a potential therapeutic for the treatment of inflammatory diseases associated with macrophage activation.
人参是五加科植物人参(C.A. Meyer)的根,是一种广为人知的传统药物,在亚洲已被使用了数千年。人参皂苷对多种疾病的控制具有多种重要的药理作用。本研究的目的是确定从人参中提取的总皂苷(TSG)对脂多糖(LPS)刺激的小鼠RAW 264.7巨噬细胞的抗炎作用。分别通过测量亚硝酸盐水平和酶联免疫吸附测定法,确定TSG对LPS诱导的一氧化氮(NO)产生以及LPS诱导的肿瘤坏死因子-α(TNF-α)和白细胞介素-1β(IL-1β)蛋白表达的抑制作用。此外,分别通过逆转录-聚合酶链反应和蛋白质印迹法,研究TSG对诱导型一氧化氮合酶(iNOS)、IL-1β和TNF-α的mRNA表达水平及定位,以及它们的上游信号蛋白,包括核因子-κB(NF-κB)和丝裂原活化蛋白激酶(MAPKs)的影响。用LPS刺激后,在RAW 264.7细胞中检测到NO产生水平升高;然而,TSG预处理通过抑制iNOS的表达,显著抑制了NO的产生(P<0.05)。此外,TSG显著降低了LPS刺激的TNF-α和IL-1β的产生(P<0.05)。在LPS刺激的RAW 264.7细胞中,NF-κB从细胞质转移到细胞核,而TSG预处理通过抑制κB抑制剂的降解,诱导NF-κB滞留在细胞质中。TSG还导致LPS刺激的RAW 264.7细胞中MAPKs的下调。这些结果表明,TSG可能具有抗炎活性,并且TSG可能被认为是治疗与巨噬细胞活化相关的炎症性疾病的潜在疗法。