title: Mechanisms underlying the antiapoptotic and anti-inflammatory effects of monotropein in hydrogen peroxide-treated osteoblasts.
abstract: Monotropein, the primary iridoid glycoside isolated from Morindacitrifolia, has been previously reported to possess potent antioxidant and antiosteoporotic properties. However, there is no direct evidence correlating the antiosteoporotic effect of monotropein with its observed antioxidant capacity, and the molecular mechanisms involved in mediating these processes remain unclear. Therefore, the aim of the present study was to investigate the protective effects of monotropein against oxidative stress in osteoblasts and the mechanisms involved in mediating this process. Osteoblast viability was evaluated using the MTT assay. The mitochondrial membrane potential and reactive oxygen species were detected by flow cytometry analyses. Western blotting and enzyme‑linked immunosorbent assays were performed to detect protein expression levels. A significant reduction in osteoblast viability was observed at 24 h following exposure to various concentrations (100‑1,000 µM) of H2O2 compared with untreated osteoblasts. The cytotoxic effect of H2O2 was notably reversed when osteoblasts were pretreated with 1‑10 µg/ml monotropein. Pretreatment with 1-10 µg/ml monotropein increased the mitochondrial membrane potential and reduced the generation of reactive oxygen species in osteoblasts following exposure to H2O2. In addition, the H2O2‑induced increase in apoptotic markers (caspase-3 and caspase-9) and H2O2-induced reduction in sirtuin 1 levels were significantly reversed following pretreatment of cells with monotropein. Furthermore, monotropein significantly reduced H2O2‑induced stimulation of NF‑κB expression, in addition to the expression of a number of proinflammatory mediators. These results indicate that monotropein suppresses apoptosis and the inflammatory response in H2O2‑induced osteoblasts through the activation of the mitochondrial apoptotic signaling pathway and inhibition of the NF‑κB signaling pathway.
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Fang-Bing
authors.initials: FB
authors.lastname: Zhu
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Jian-Yue
authors.initials: JY
authors.lastname: Wang
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Ying-Liang
authors.initials: YL
authors.lastname: Zhang
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Yun-Gen
authors.initials: YG
authors.lastname: Hu
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Zhen-Shuang
authors.initials: ZS
authors.lastname: Yue
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Lin-Ru
authors.initials: LR
authors.lastname: Zeng
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Wen-Jie
authors.initials: WJ
authors.lastname: Zheng
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Qiao
authors.initials: Q
authors.lastname: Hou
authors.affiliation: Department of Orthopedic Surgery, The Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou, Zhejiang 310009, P.R. China.
authors.forename: Shi-Gui
authors.initials: SG
authors.lastname: Yan
authors.affiliation: Department of Orthopedic Surgery, Xiaoshan Traditional Chinese Medical Hospital, Hangzhou, Zhejiang 311200, P.R. China.
authors.forename: Ren-Fu
authors.initials: RF
authors.lastname: Quan
mesh_terms: D000469:Alkaline Phosphatase / Q000378:metabolism; D000818:Animals; D000893:Anti-Inflammatory Agents / Q000494:pharmacology*; D000975:Antioxidants / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects*; D002454:Cell Differentiation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D051546:Cyclooxygenase 2 / Q000235:genetics / Q000378:metabolism; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D039823:Iridoids / Q000494:pharmacology*; D008297:Male; D020782:Matrix Metalloproteinases / Q000378:metabolism; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D016328:NF-kappa B / Q000378:metabolism; D052247:Nitric Oxide Synthase Type II / Q000235:genetics / Q000378:metabolism; D010006:Osteoblasts / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism*; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017382:Reactive Oxygen Species / Q000378:metabolism
chemical_list: D000893:Anti-Inflammatory Agents; D000975:Antioxidants; D039823:Iridoids; D016328:NF-kappa B; D017382:Reactive Oxygen Species; C507582:monotropein; D006861:Hydrogen Peroxide; D052247:Nitric Oxide Synthase Type II; D051546:Cyclooxygenase 2; D000469:Alkaline Phosphatase; D020782:Matrix Metalloproteinases