Kang Sera, Siddiqi Muhammad Hanif, Yoon Sung Joo, Ahn Sungeun, Noh Hae-Yong, Kumar Natarajan Sathish, Kim Yeon-Ju, Yang Deok-Chun
Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
In Vitro Cell Dev Biol Anim. 2016 Oct;52(9):895-905. doi: 10.1007/s11626-016-0062-9. Epub 2016 Jul 1.
Ginsenosides have been used traditionally as an oriental medicine. However, the anti-osteoarthritic effect of ginsenoside compound K (hereafter referred to as CK) has not been reported. Therefore, in this study, the protective effects of CK were evaluated in silico and in vitro using HO-stimulated MC3T3-E1 cells by measuring the levels of proinflammatory cytokines responsible for articular cartilage degradation. In silico results demonstrated that, among the selected ginsenosides, CK is a non-toxic drug-like molecule with strong binding affinity for selected cytokine-activated kinase such as IkBα kinase (IKK). The molecular binding energy of CK with the active sites of IKK suggests anti-osteoarthritic functions. Cultured HO-stimulated MC3T3-E1 cells that were exposed to CK showed dramatically increased expression of osteoblast differentiation markers such as alkaline phosphatase (ALP) activity, type I collagen (Col-I) content, and mineralization. During aging, HO also leads to the production of reactive oxygen species (ROS) and nitric oxide (NO), which play important roles in the development of osteoarthritis (OA). Therefore, the effect of CK on ROS and NO generation was also examined. Our results showed that CK dose-dependently inhibited HO-induced ROS and NO production in MC3T3-E1 cells. Moreover, qRT-PCR data showed that CK increased expression of osteogenic markers such as ALP and Col-I but decreased expression of inflammatory-related genes including IKK and interleukin 1β (IL-1β) in a dose-dependent manner in HO-stimulated MC3T3-E1 cells. The findings of this study suggest the use of CK as a novel protective and therapeutic agent in AO.
人参皂苷传统上一直被用作一种东方药物。然而，人参皂苷化合物K（以下简称CK）的抗骨关节炎作用尚未见报道。因此，在本研究中，通过测量负责关节软骨降解的促炎细胞因子水平，在计算机模拟和体外实验中使用过氧化氢（HO）刺激的MC3T3-E1细胞评估了CK的保护作用。计算机模拟结果表明，在所选的人参皂苷中，CK是一种无毒的类药物分子，对所选的细胞因子激活激酶如IkBα激酶（IKK）具有很强的结合亲和力。CK与IKK活性位点的分子结合能表明其具有抗骨关节炎功能。暴露于CK的培养的HO刺激的MC3T3-E1细胞显示成骨细胞分化标志物的表达显著增加，如碱性磷酸酶（ALP）活性、I型胶原蛋白（Col-I）含量和矿化。在衰老过程中，HO还会导致活性氧（ROS）和一氧化氮（NO）的产生，它们在骨关节炎（OA）的发展中起重要作用。因此，还研究了CK对ROS和NO生成的影响。我们的结果表明，CK在MC3T3-E1细胞中剂量依赖性地抑制HO诱导的ROS和NO产生。此外，qRT-PCR数据显示，在HO刺激的MC3T3-E1细胞中，CK剂量依赖性地增加成骨标志物如ALP和Col-I的表达，但降低包括IKK和白细胞介素1β（IL-1β）在内的炎症相关基因的表达。本研究结果表明CK可作为AO的一种新型保护和治疗剂。
