Lee Byung-Hwan, Choi Sun-Hye, Kim Hyeon-Joong, Jung Seok-Won, Kim Hyunsook, Shin Ho-Chul, Lee Joon-Hee, Hwang Sung-Hee, Kim Hyoung-Chun, Nah Seung-Yeol
Department of Physiology, College of Veterinary Medicine and Bio/Molecular Informatics Center, Konkuk University.
Biol Pharm Bull. 2015;38(10):1631-7. doi: 10.1248/bpb.b15-00171.
Gintonin is a novel ginseng-derived G protein-coupled lysophosphatidic acid (LPA) receptor ligand. Gintonin elicits an [Ca(2+)]i transient in animal cells via activation of LPA receptors. In vitro studies have shown that gintonin regulates various calcium-dependent ion channels and receptors. In in vivo studies, gintonin elicits anti-Alzheimer's disease activity through the activation of the non-amyloidogenic pathway and anti-metastatic effects through the inhibition of autotaxin. However, a method for gintonin quantitation in ginseng has not been developed. In the present study, we developed an enzyme immunoassay (EIA) to measure gintonin. A monoclonal antibody was raised in a mouse using gintonin as the immunogen, and an indirect competitive EIA was used to measure gintonin. The working range was 0.01-10 µg per assay. The anti-gintonin monoclonal antibody did not cross-react with the ginsenosides Ra, Rb1, Rb2, Rc, Rd, Re, Rf, Rg1, and Rg3 or with LPAs such as LPA C16:0, LPA C18:0, LPA C18:1, and LPA C18:2. Using a standard curve, we measured the amount of gintonin in various ginseng extract fractions. Interestingly, we only detected a little amount of gintonin in conventional hot water extracts of Korean red ginseng. However, we can measure gintonin after ethanol extraction of Korean red ginseng marc. Thus, gintonin can be extracted from ginseng with ethanol but not water, and the remaining Korean red ginseng marc can be used to obtain gintonin. These results indicate that the EIA with the anti-gintonin monoclonal antibody can be used to quantify gintonin in various ginseng preparations, including commercial ginseng products.
人参皂草苷是一种新型的源自人参的G蛋白偶联溶血磷脂酸（LPA）受体配体。人参皂草苷通过激活LPA受体在动物细胞中引发细胞内钙离子浓度（[Ca(2+)]i）瞬变。体外研究表明，人参皂草苷可调节多种钙依赖性离子通道和受体。在体内研究中，人参皂草苷通过激活非淀粉样蛋白生成途径发挥抗阿尔茨海默病活性，并通过抑制自分泌运动因子发挥抗转移作用。然而，尚未开发出人参中人参皂草苷的定量方法。在本研究中，我们开发了一种酶免疫分析（EIA）方法来测定人参皂草苷。以人参皂草苷作为免疫原在小鼠体内制备单克隆抗体，并使用间接竞争EIA法测定人参皂草苷。每次测定的工作范围为0.01 - 10μg。抗人参皂草苷单克隆抗体与人参皂苷Ra、Rb1、Rb2、Rc、Rd、Re、Rf、Rg1和Rg3或与LPA如LPA C16:0、LPA C18:0、LPA C18:1和LPA C18:2均无交叉反应。使用标准曲线，我们测定了各种人参提取物组分中人参皂草苷的含量。有趣的是，我们在韩国红参的传统热水提取物中仅检测到少量人参皂草苷。然而，对韩国红参药渣进行乙醇提取后，我们能够测定人参皂草苷。因此，人参皂草苷可用乙醇而非水从人参中提取，剩余的韩国红参药渣可用于获取人参皂草苷。这些结果表明，使用抗人参皂草苷单克隆抗体的EIA法可用于定量各种人参制品（包括市售人参产品）中的人参皂草苷。
