长链非编码RNA NEAT1在骨质疏松症中的表达水平及对成骨细胞和破骨细胞分化的影响研究  

Expression of long-chain non-coding RNA NEAT1 in osteoporosis and its effect on differentiation of osteoblasts and osteoclasts

作　　者：樊萍[1] 冯秀媛[1] 胡楠 蒲丹[1] 吕晓虹[1] 孙怡宁[1] 何岚[1] FAN Ping;FENG Xiu-yuan;HU Nan(Department of Rheumatology,The First Affiliated Hospital of Xi'an Jiaotong University,Xi'an Shaanxi 710061,China)

机构地区：[1]西安交通大学第一附属医院风湿免疫科

出　　处：《临床和实验医学杂志》2019年第23期2504-2507,共4页Journal of Clinical and Experimental Medicine

基　　金：陕西省国际科技合作与交流计划项目(编号:2016KW-023)

摘　　要：目的研究长链非编码RNA NEAT1(lncRNA-NEAT1)在骨质疏松症中的表达水平及对破骨细胞与成骨细胞分化的影响。方法通过鼠尾悬吊法制备小鼠骨质疏松症模型,设置实验组与对照组。通过实时荧光定量聚合酶链反应检测lncRNA-NEAT1在小鼠成骨细胞系MC3T3-E1向成骨分化过程中的表达量,测定lncRNA-NEAT1在小鼠巨噬细胞系RAW264.7向破骨分化过程中的表达量;取敲低lncRNA-NEAT1慢病毒感染的前成骨细胞系MC3T3-E1,通过碱性磷酸酶染色观察成骨细胞的活性情况;取敲低lncRNA-NEAT1慢病毒感染的巨噬细胞系RAW264.7,通过抗酒石酸酸性磷酸酶染色明确破骨细胞的活性情况。结果实验组小鼠股骨组织中lncRNA-NEAT1的相对表达量明显高于对照组(P<0.01),巨噬细胞系RAW264.7破骨分化过程中的lncRNA-NEAT1表达量较对照组显著上调(P<0.01),前成骨细胞系MC3T3-E1向成骨分化过程中的lncRNA-NEAT1表达量较对照组显著下调(P<0.01)。经碱性磷酸酶染色可知,通过敲低NEAT1的表达可明显提高成骨细胞活性;经抗酒石酸酸性磷酸酶染色可知,通过敲低NEAT1的表达可明显抑制破骨细胞的活性。结论lncRNA-NEAT1在成骨分化过程中的表达显著下调,而在破骨分化过程中的表达显著上调,提示干扰lncRNA-NEAT1的表达可通过诱导破骨细胞分化和阻滞成骨细胞分化,从而而导致骨质疏松症的发生。Objective To study the expression of long non-coding RNA NEAT1(lncRNA-NEAT1)in osteoporosis and its effect on the differentiation of osteoclasts and osteoblasts.Methods Osteoporosis model of mice was established by tail suspension method.The expression of lncRNA-NEAT1 during osteogenic differentiation of mouse osteoblast line MC3T3-E1 was detected by real-time fluorescence quantitative polymerase chain reaction.The osteoclast differentiation of mouse macrophage line RAW264.7 was measured by lncRNA-NEAT1.The activity of osteoclasts was observed by alkaline phosphatase staining,and the activity of osteoclasts was determined by tartrate-resistant acid phosphatase staining in RAW264.7 macrophage line infected with lncRNA-NEAT1 lentivirus.Results The relative expression of lncRNA-NEAT1 in femoral tissue of experimental group was significantly higher than that of control group(P<0.01).The expression of lncRNA-NEAT1 in osteoclast differentiation of RAW264.7 macrophage line was significantly higher than that of control group(P<0.01).The expression of lncRNA-NEAT1 in osteoblast differentiation of MC3T3-E1 was significantly higher than that of control group(P<0.01).There was significant decrease(P<0.01).Alkaline phosphatase staining showed that the activity of osteoblasts could be significantly increased by knocking down the expression of NEAT1.Tartaric acid phosphatase staining showed that knocking down the expression of NEAT1 could significantly inhibit the activity of osteoclasts.Conclusion The expression of lncRNA-NEAT1 is significantly down-regulated in the process of site differentiation and up-regulated in the process of osteoclast differentiation,suggesting that interfering with the expression of lncRNA-NEAT1 can induce osteoclast differentiation and block osteoblast differentiation,thus leading to osteoporosis.

关 键 词：小鼠 骨质疏松症 长链非编码RNA NEAT1 破骨细胞 成骨细胞 细胞分化 

分 类 号：R73[医药卫生—肿瘤]