四烯甲萘醌保护成骨细胞氧化损伤的作用研究  

Study on the protective effect of menatetrenone against the oxidative stress of osteoblasts

作　　者：蒋益忠 林丽娟 辛海量 金玉娥[3] 蒋益萍 薛黎明[3] JIANG Yizhong;LIN Lijuan;XIN Hailiang;JIN Yu’e;JIANG Yiping;XUE Liming(Dongyang Traditional Chinese Medicine Hospital of Zhejiang,Dongyang 322105,China;Department of Pharmacognosy,School of Pharmacy,Naval Medical University,Shanghai 200433,China;Institute of Chemical Toxicity,Shanghai Municipal Center for Disease Control and Prevention,Shanghai 200336,Chian)

机构地区：[1]浙江省东阳市中医院,浙江东阳322105 [2]海军军医大学药学院生药学教研室,上海200433 [3]上海市疾病预防控制中心化学品毒性检定所,上海200336

出　　处：《药学实践杂志》2020年第6期523-527,共5页Journal of Pharmaceutical Practice

基　　金：上海市卫生和计划生育委员会面上项目(201740068);海军军医大学本科创新孵化基地项目(PH2019102)。

摘　　要：目的考察四烯甲萘醌(MK4)对成骨细胞氧化损伤的保护作用,阐明MK4防治骨质疏松作用机制。方法采用过氧化氢(H2O2)刺激小鼠成骨细胞系(MC3T3-E1)氧化应激模型,考察细胞活力、ALP活性和骨结节面积,DCFH-DA法检测活性氧(ROS)水平,JC-1检测线粒体膜电势,Annexin V-FITC/PI法检测细胞凋亡率,RT-PCR法考察氧化应激相关基因FoxO1、FoxO3、SOD、Bcl-2和bax等的mRNA表达。结果10μmol/L四烯甲萘醌能显著提高H2O2刺激的成骨细胞增殖、ALP活性、骨结节形成面积和增强细胞膜电势,显著降低H2O2刺激的成骨细胞内丙二醛和活性氧水平,同时显著降低成骨细胞凋亡率和细胞凋亡因子bax/Bcl-2的mRNA表达水平,显著提高抗氧化酶SOD和转录因子FoxO1、FoxO3的mRNA表达。结论四烯甲萘醌可通过调控FoxO通路保护成骨细胞氧化损伤和通过下调bax/Bcl-2比例,降低成骨细胞凋亡。Objective To investigate the protective effect of menatetrenone(MK4)on the osteoblasts in oxidative stress,and to clarify the anti-osteoporosis mechanism of MK4.Methods Mouse osteoblasts(MC3T3-E1)induced by hydrogen peroxide(H2O2)was used.Cell viability,ALP activity and the area of bone nodule were observed.The level of ROS was detected by DCFHDA,mitochondrial membrane potential by JC-1,apoptosis rate by annexin V-FITC/PI,and the expression of FoxO1,FoxO3,SOD,bcl-2 and bax by RT-PCR.Results Menatetrenone at 10μmol/L significantly increased the proliferation of osteoblasts stimulated by H2O2,ALP activity,bone nodule formation area,cell membrane potential,the antioxidant SOD and transcription factors FoxO1 and FoxO3 mRNA expression.In the meantime,the elevated malondialdehyde and reactive oxygen species level in cells induced by H2O2,the apoptosis rate and the mRNA expression level of bax/Bcl-2 were significantly reduced.Conclusion menatetrenone can protect osteoblasts from oxidative damage by regulating FoxO pathway and reduce osteoblasts apoptosis by up regulating the proportion of Bcl-2/bax.

关 键 词：四烯甲萘醌 骨质疏松 成骨细胞 氧化损伤 FoxO通路 

分 类 号：R15[医药卫生—营养与食品卫生学]