罗汉果苷V促进LncRNA TUG1表达刺激成骨细胞的增殖与分化  

Mogroside V stimulates osteoblast proliferation and differentiation by promoting LncRNA TUG1 expression

作　　者：姚顺晗 韦华成 覃家港 廖亮[1] Yao Shunhan;Wei Huacheng;Qin Jiagang;Liao Liang(First Affiliated Hospital of Guangxi Medical University,Nanning 530021,Guangxi Zhuang Autonomous Region,China)

机构地区：[1]广西医科大学第一附属医院,广西壮族自治区南宁市530021

出　　处：《中国组织工程研究》2020年第26期4129-4134,共6页Chinese Journal of Tissue Engineering Research

基　　金：广西自然科学基金面上项目(2018GXNSFAA138078),项目负责人:廖亮。

摘　　要：背景:骨质疏松是骨在进行重塑的过程中,成骨细胞的骨形成和破骨细胞的骨吸收之间的动态平衡紊乱所致。在细胞水平严格控制骨重建,对于维持骨稳态和避免发生骨质疏松有重要意义。前期研究表明1.25×10^(-2)g/L的罗汉果苷V可促进成骨细胞增殖、分化,其机制可能涉及Lnc RNA TUG1。目的:探讨Lnc RNA TUG1在罗汉果苷V促进成骨细胞增殖与分化中的作用。方法:采用两步酶消化法提取SD乳鼠成骨细胞,随后将细胞分为2组,分别给予0,1.25×10^(-2)g/L的罗汉果苷V干预,干预培养2 d后进行Lnc RNA基因检测;设计并合成Lnc RNA TUG1沉默病毒,将新提取的成骨细胞分为正常细胞对照组、罗汉果苷V干预组、罗汉果苷V+阴性病毒组、TUG1沉默组、罗汉果苷V+TUG1沉默组。按实验分组对成骨细胞进行干预处理,随后分别在2,4,6 d后通过FDA荧光染色观察成骨细胞增殖活性,干预6d后通过碱性磷酸酶染色、茜素红染色及q RT-PCR检测TUG1在罗汉果苷V促进成骨细胞增殖与分化中的作用。结果与结论:(1)Lnc RNA基因检测表明1.25×10^(-2)g/L的罗汉果皂苷显著促进成骨细胞内Lnc RNA TUG1表达;(2)FDA荧光染色显示TUG1沉默可抑制罗汉果苷V促进成骨细胞增殖;(2)干预6 d后,碱性磷酸酶染色及茜素红染色实验均显示TUG1沉默可抑制罗汉果苷V促进成骨细胞成骨矿化;(3)q RT-PCR结果显示成骨分化相关基因RUNX2、BSP、OCN和COL1A1在罗汉果苷V干预组显著高表达,而在罗汉果苷V干预+TUG1沉默组则表达受抑制;(4)以上结果表明,罗汉果苷V通过促进Lnc RNATUG1表达刺激成骨细胞的增殖与分化。BACKGROUND:Osteoporosis is a balance disorder between bone formation of osteoblasts and bone resorption of osteoclasts during bone remodeling.Strict control of bone remodeling at the cellular level is important to maintain bone homeostasis and avoid osteoporosis.Previous studies have shown that 1.25×10-2 g/L mogroside V can promote osteoblast proliferation and differentiation,and its mechanism may be related to LncRNA TUG1.OBJECTIVE:To investigate the role of LncRNA TUG1 in the promotion of osteoblast proliferation and differentiation by mogroside V.METHODS:Osteoblasts from neonatal Sprague-Dawley rats were extracted by two-step enzymatic digestion.The cells were divided into two groups and treated with 0 and 1.25×10-2 g/L Mogroside V.The LncRNA was detected after 2 days of culture.LncRNA TUG1 silencing virus was designed and synthetized.The newly extracted osteoblasts were divided into normal cell control group,mogroside V intervention group,mogroside V+negative virus group,TUG1 silent group,and mogroside V+TUG1 silent group.The proliferation of osteoblasts was observed by FDA fluorescence staining at 2,4,and 6 days after processing according to the above grouping conditions.After 6 days of treatment on osteoblasts,the effect of TUG1 on osteoblast proliferation and differentiation was studied by alkaline phosphatase staining,alizarin red staining and qRT-PCR.RESULTS AND CONCLUSION:LncRNA detection showed that 1.25×10-2 g/L Mogroside V significantly promoted the expression of LncRNA TUG1 in osteoblasts.FDA fluorescent staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on osteoblast proliferation.After 6 days of treatment,alkaline phosphatase staining and alizarin red staining showed that silencing of TUG1 inhibited the positive effect of mogroside V on mineralization of osteoblasts.The results of qRT-PCR showed that Runx2,BSP,OCN and COL1 A1 genes were highly expressed in the mogroside V intervention group,but their expression was inhibited in the mogroside V+TUG1 silent group.

关 键 词：罗汉果苷V LncRNA TUG1 成骨细胞 细胞增殖 细胞分化 骨质疏松 

分 类 号：R446[医药卫生—诊断学] R496[医药卫生—临床医学]