绞股蓝皂苷减轻H2O2诱导大鼠成骨细胞氧化应激损伤的机制  

Mechanism by which gypenosides alleviate oxidative stress injury induced by H2O2 in rat osteoblasts

作　　者：林燕平 黄佳纯 陈桐莹 马江涛 郭海威 汪悦东 袁嘉尧 姜涛[1,3] 黄宏兴 黄红[1,4] 万雷 Lin Yanping;Huang Jiachun;Chen Tongying;Ma Jiangtao;Guo Haiwei;Wang Yuedong;Yuan Jiayao;Jiang Tao;Huang Hongxing;Huang Hong;Wan Lei(Guangzhou University of Chinese Medicine,Guangzhou 510000,Guangdong Province,China;Lingnan Medical Research Center,Guangzhou 510080,Guangdong Province,China;Third Affiliated Hospital of Guangzhou University of Chinese Medicine,Guangzhou 510240,Guangdong Province,China;School of Nursing,Guangzhou University of Chinese Medicine,Guangzhou 510000,Guangdong Province,China)

机构地区：[1]广州中医药大学,广东省广州市510000 [2]岭南医学研究中心,广东省广州市510080 [3]广州中医药大学第三附属医院,广东省广州市510240 [4]广州中医药大学护理学院,广东省广州市510000

出　　处：《中国组织工程研究》2020年第23期3649-3653,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金面上项目(81973886,81674004),项目负责人:黄宏兴;广州中医药大学学科研究重点项目(XK2019028),项目负责人:黄宏兴。

摘　　要：背景:绞股蓝皂苷具有抗氧化作用,多应用于降血压、抗衰老、抗肿瘤等,但其具体保护机制尚不明确,对氧化应激损伤的成骨细胞增殖、分化的影响亦未知。目的:探讨绞股蓝皂苷减轻大鼠成骨细胞氧化应激损伤的机制,以及对氧化损伤的成骨细胞的增殖、分化的影响。方法:采用分离乳鼠颅骨细胞的单层培养法培养成骨细胞,实验分为3组,以普通生长培养基为空白组,以普通生长培养基+氧化损伤为对照组,以含绞股蓝皂苷的培养基+氧化损伤为实验组。实验组和对照组成骨细胞经含150μmol/L H2O2的生长培养基诱导氧化损伤模型。干预3,5 d CCK8法检测绞股蓝皂苷对氧化损伤的成骨细胞活性的影响;在诱导后第7天采用碱性磷酸酶染色检测成骨细胞碱性磷酸酶的活性;诱导第21天采用茜素红染色观察成骨细胞矿化情况;Western blot检测NOX4、骨形态发生蛋白2和Smad4的蛋白表达量。实验方案经广州中医药大学动物实验伦理委员会批准。结果与结论:①绞股蓝皂苷可促进氧化损伤的成骨细胞增殖;②碱性磷酸酶染色和茜素红染色结果表明绞股蓝皂苷对受氧化应激损伤的成骨细胞的分化有一定促进作用;③与对照组相比,绞股蓝皂苷可下调实验组NOX4蛋白的表达,上调实验组骨形态发生蛋白2和Smad4蛋白的表达(P<0.05);④结果说明,绞股蓝皂苷对H2O2诱导的成骨细胞氧化应激损伤具有一定的保护作用,并且对损伤的成骨细胞有促进增殖分化的作用,其机制可能与抑制NOX4蛋白表达并激活BMP/Smad通路有关。BACKGROUND:Gypenosides have antioxidant properties,with beneficial effects such as reducing blood pressure,anti-aging and anti-tumor,but the specific protective mechanism is not clear.It is also unknown whether gypenosides have effect on the proliferation and differentiation of oxidative stress-damaged osteoblasts.OBJECTIVE:To investigate the mechanism by which gypenosides alleviate oxidative stress injury in rat osteoblasts and the effect on the proliferation and differentiation of oxidatively damaged osteoblasts.METHODS:Monolayer cell culture method was used to separate neonatal rat skull cells for the culture of osteoblasts.In this experiment,there were three groups,with normal culture medium as blank group,normal culture medium+oxidative damage as control group,and normal culture medium containing gypenosides and oxidative damage as experimental group.Osteoblasts in the experimental and control groups were cultured in the culture medium containing 150μmol/L H2O2.After 3 and 5 days of intervention,cell counting kit-8 method was used to detect the effects of gypenosides on oxidative damage of osteoblasts.Alkaline phosphatase staining was used to detect alkaline phosphatase activity on day 7 after induction.Alizarin red staining was used on day 21 of induction to observe osteoblast mineralization.Western blot was used to detect the expression of NOX4,bone morphogenetic protein 2 and Smad4.The experimental protocol was approved by the Animal Ethics Committee of Guangzhou University of Chinese Medicine.RESULTS AND CONCLUSION:Gypenosides could promote the proliferation of oxidatively damaged osteoblasts.The results of alkaline phosphatase staining and alizarin red staining showed that gypenosides could promote the differentiation of oxidatively damaged osteoblasts.Compared with the control group,gypenosides could downregulate the expression of NOX4 protein and upregulate the expression of bone morphogenetic protein 2 and Smad4 protein in the experimental group,with statistically significant results(P<0.05).All the

关 键 词：骨质疏松 成骨细胞 绞股蓝皂苷 氧化应激 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]