机构：[1]Laboratory of Molecular Neuropathology, Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou 215021, China[2]Department of Pharmacy, Suzhou Hospital Affiliated with Nanjing Medical University, Suzhou 215002, China[3]The Second Affiliated Hospital of Soochow University, Soochow University, Suzhou 215021, China
Aim: Increasing evidence has shown that environmental factors such as rotenone and paraquat induce neuroinflammation, which contributes to the pathogenesis of Parkinson's disease (PD). In this study, we investigated the molecular mechanisms underlying the repression by menaquinone-4 (MK-4), a subtype of vitamin K-2, of rotenone-induced microglial activation in vitro. Methods: A microglial cell line (BV2) was exposed to rotenone (1 mu mol/L) with or without MK-4 treatment. The levels of TNF-alpha or IL-1 beta in 100 mu L of cultured media of BV2 cells were measured using ELISA kits. BV2 cells treated with rotenone with or without MK4 were subjected to mitochondrial membrane potential, ROS production, immunofluorescence or immunoblot assays. The neuroblastoma SH-SY5Y cells were treated with conditioned media (CM) of BV2 cells that were exposed to rotenone with or without MK-4 treatment, and the cell viability was assessed using MTT assay. Results: In rotenone-treated BV2 cells, MK-4 (0.5-20 mu mol/L) dose-dependently suppressed the upregulation in the expression of iNOS and COX-2 in the cells, as well as the production of TNF-alpha and IL-1 beta in the cultured media. MK-4 (5-20 mu mol/L) significantly inhibited rotenone-induced nuclear translocation of NF-kappa B in BV2 cells. MK-4 (5-20 mu mol/L) significantly inhibited rotenone-induced p38 activation, ROS production, and caspase-1 activation in BV2 cells. MK-4 (5-20 mu mol/L) also restored the mitochondrial membrane potential that had been damaged by rotenone. Exposure to CM from rotenone-treated BV2 cells markedly decreased the viability of SH-SY5Y cells. However, this rotenone-activated microglia-mediated death of SH-SY5Y cells was significantly attenuated when the BV2 cells were co-treated with MK-4 (5-20 mu mol/L). Conclusion: Vitamin K2 can directly suppress rotenone-induced activation of microglial BV2 cells in vitro by repressing ROS production and p38 activation.
基金：
This work was supported in part by the National Natural Sci-ences Foundation of China (No 31330030), the National High-tech Research and Development program of China 973-proj-ects (No 2012CB947602), and the National Natural Sciences Foundation of China (Nos 81371393 and 31300887), a Project Funded by the Priority Academic Program Development of Jiangsu Higher Education Institutions, Suzhou Clinical Research Center of Neurological Disease (No Szzx201503) and Jiangsu Provincial Special Program of Medical Science (No BL2014042).
第一作者机构：[1]Laboratory of Molecular Neuropathology, Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou 215021, China[2]Department of Pharmacy, Suzhou Hospital Affiliated with Nanjing Medical University, Suzhou 215002, China
通讯作者：
通讯机构：[1]Laboratory of Molecular Neuropathology, Jiangsu Key Laboratory of Translational Research and Therapy for Neuro-Psycho-Diseases and College of Pharmaceutical Sciences, Soochow University, Suzhou 215021, China[3]The Second Affiliated Hospital of Soochow University, Soochow University, Suzhou 215021, China
医学