title: Therapeutic potential of compound K as an IKK inhibitor with implications for osteoarthritis prevention: an in silico and in vitro study.
abstract: 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 H2O2-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 H2O2-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, H2O2also 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 H2O2-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 H2O2-stimulated MC3T3-E1 cells. The findings of this study suggest the use of CK as a novel protective and therapeutic agent in AO.
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Sera
authors.initials: S
authors.lastname: Kang
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Muhammad Hanif
authors.initials: MH
authors.lastname: Siddiqi
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Sung Joo
authors.initials: SJ
authors.lastname: Yoon
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Sungeun
authors.initials: S
authors.lastname: Ahn
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Hae-Yong
authors.initials: HY
authors.lastname: Noh
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Natarajan Sathish
authors.initials: NS
authors.lastname: Kumar
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea.
authors.forename: Yeon-Ju
authors.initials: YJ
authors.lastname: Kim
authors.affiliation: Graduate School of Biotechnology, College of Life Science, Kyung Hee University, Seocheon, Giheunggu Yonginsi, Gyeonggido, 449-701, South Korea. dcyang@khu.ac.kr.
authors.forename: Deok-Chun
authors.initials: DC
authors.lastname: Yang
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000818:Animals; D002113:Calcification, Physiologic / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003094:Collagen / Q000096:biosynthesis; D003198:Computer Simulation; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics / Q000378:metabolism; D004353:Drug Evaluation, Preclinical; D036145:Ginsenosides / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D006861:Hydrogen Peroxide / Q000633:toxicity; D051550:I-kappa B Kinase / Q000037:antagonists & inhibitors* / Q000378:metabolism; D053583:Interleukin-1beta / Q000235:genetics / Q000378:metabolism; D008810:Mice, Inbred C57BL; D062105:Molecular Docking Simulation; D009569:Nitric Oxide / Q000096:biosynthesis; D010003:Osteoarthritis / Q000188:drug therapy* / Q000473:pathology / Q000517:prevention & control*; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects / Q000201:enzymology; D015675:Osteocalcin / Q000235:genetics / Q000378:metabolism; D047428:Protein Kinase Inhibitors / Q000737:chemistry / Q000494:pharmacology / Q000627:therapeutic use*; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism
chemical_list: D000469:Alkaline Phosphatase; D003094:Collagen; D050659:Core Binding Factor Alpha 1 Subunit; D036145:Ginsenosides; D006861:Hydrogen Peroxide; D051550:I-kappa B Kinase; D053583:Interleukin-1beta; D009569:Nitric Oxide; D015675:Osteocalcin; D047428:Protein Kinase Inhibitors; D012333:RNA, Messenger; C112772:ginsenoside compound K
keywords: Differentiation; Ginsenosides; IKK/NF-kB; MC3T3-E1 cells; Osteoarthritis; Osteoblasts