摘要
目的探讨白藜芦醇对脂多糖(LPS)诱导的MC3T3-E1成骨细胞的骨保护作用。方法采用CCK-8法检测白藜芦醇不同浓度下MC3T3-E1成骨细胞的细胞增殖活性,对硝基苯磷酸盐法检测白藜芦醇不同浓度下MC3T3-E1成骨细胞ALP活性。将MC3T3-E1成骨细胞分为空白组(基础培养基)、对照组(LPS 2μg/mL)和实验组(LPS 2μg/mL+白藜芦醇20μmol/L)。采用qRT-PCR检测三组成骨相关基因Runx2、ALP、骨钙素(OCN)和骨桥蛋白(OPN) mRNA表达,Western blot法检测三组细胞沉默调节蛋白1(SIRT1)蛋白表达。结果白藜芦醇20μmol/L是MC3T3-E1成骨细胞最适成骨浓度(P<0.05)。与空白组比较,实验组成骨相关基因Runx2、ALP、OCN和OPN mRNA表达水平和SIRT1蛋白表达水平均降低(P<0.05);与对照组相比,实验组成骨相关基因Runx2、ALP、OCN和OPN mRNA表达水平和SIRT1蛋白表达水平升高(P<0.05)。结论白藜芦醇能够通过提高Runx2、ALP、OCN、OPN和SIRT1相关成骨因子的表达对LPS诱导的MC3T3-E1成骨细胞发挥骨保护作用。
Objective To investigate the bone protective effect of resveratrol on lipopolysaccharide(LPS)-induced MC3 T3-E1 osteoblasts.Methods Proliferation activity of MC3 T3-E1 osteoblasts in different concentrations of resveratrol was detected by CCK-8.ALP activity of MC3 T3-E1 osteoblasts in different concentrations of resveratrol was determined by pNPP method.MC3 T3-E1 osteoblasts were divided into three groups of A(treated with LPS 2 μg/mL + resveratrol 20 μmol/L),B(treated with LPS 2 μg/mL) and C(blank control).qRT-PCR was used to detect the mRNA expressions of osteogenic genes Runx2,ALP,osteocalcin(OCN) and osteopontin(OPN) in three groups.The protein expression of SIRT1 in three groups was detected by Western blot.Results Resveratrol 20 μmol/L was the optimal osteogenic concentration for MC3 T3-E1 osteoblasts(P<0.05).Compared with group C,mRNA expression levels of Runx2,ALP,OCN and OPN and SIRT1 protein expression level were decreased in group A(P<0.05).Compared with group B,mRNA expression levels of Runx2,ALP,OCN and OPN and SIRT1 protein expression level were increased in group A(P<0.05).Conclusion Resveratrol can play a bone protective role in LPS-induced MC3 T3-E1 osteoblastic cells by increasing the expression of Runx2,ALP,OCN,OPN and SIRT1-related osteoblastic factors.
作者
刘子歌
田佳宁
袁衡
李昃鹏
张晨
宋国瑞
陈德胜
LIU Zige;TIAN Jianing;YUAN Heng(Department of Orthopedics,General Hospital of Ningxia Medical University,Yinchuan 75000A,CHINA)
出处
《江苏医药》
CAS
2021年第3期222-226,共5页
Jiangsu Medical Journal
基金
国家自然科学基金(82060408、81760405、81560364)
宁夏自然科学基金重点项目(2018AAC02013)
宁夏医科大学校级课题重点项目(XZ2018014)。
关键词
白藜芦醇
脂多糖
成骨细胞
Resveratrol
Lipopolysaccharide
Osteoblasts
