lncRNA NEAT1过表达促进破骨细胞分化并抑制成骨细胞分化诱发骨质疏松

Over-expression of long noncoding RNA NEAT1 induces osteoporosis via promoting osteoclaste differentiation and inhibiting osteoblast differentiation

目的:研究长链非编码RNA NEAT1在骨质疏松症中的表达及对成骨细胞和破骨细胞分化的作用。方法:收集正常人和骨质疏松症患者的骨组织,运用Real-time PCR检测NEAT1的表达水平。在卵巢去势(OVX)和鼠尾悬吊(TS)诱导的小鼠疏松股骨中,在小鼠前成骨细胞系MC3T3-E1和人间充质干细胞h MSC向成骨分化过程中以及小鼠巨噬细胞系RAW264.7向破骨分化过程中,检测NEAT1的表达水平。利用敲低Neat1的慢病毒感染MC3T3-E1和RAW264.7细胞,分别采用碱性磷酸酶(ALP)染色确定成骨细胞活性以及抗酒石酸酸性磷酸酶(TRAP)染色观察破骨细胞活性变化。结果:NEAT1在患者疏松的骨组织中、小鼠的疏松股骨组织中以及RAW264.7细胞破骨分化过程中表达明显增多;而在MC3T3-E1和hMSC成骨分化过程中表达明显降低。ALP染色显示敲低Neat1显著加重成骨细胞活性,TRAP染色显示敲低Neat1显著抑制破骨细胞活性。结论:长链非编码RNA NEAT1在成骨分化过程中低表达,在破骨分化过程中过表达,并通过促进破骨细胞分化及抑制成骨细胞分化诱发骨质疏松。

Objective: To examine the expression of lncRNA NEAT1 in osteoporotic bone tissue and its effect on the osteoclaste andosteoblast differentiation. Methods: The transcript of NEAT1 in human normal and osteoporotic bone tissues was carried out;ovariectomized (OVX) and hindlimb -unloaded induced osteoporosis in the femurs of mice;process of hMSCs and MC3T3 -E1 cells differentiating to osteoblast and the process of RAW264.7 cells differentiating toosteoclast were detected by Real-time PCR,respectively. Moreover,when NEAT1 was silenced,the activity of osteoblast was determined by ALP staining and the activity of osteoclast was determined by TRAP staining. Results: NEAT1 was significantly up-regulated in the osteoporotic bone tissues of the patients and the osteoporotic mice models as well as in the process of osteoclast differentiation.Furthermore,NEAT1 was down -regulated in the process ofosteoblast differentiation.In addition,when NEAT1 was silenced,the expression of Ctsk and the activity of osteoclast were decreased while the expression of Alp and the activity of osteoblast were increased. Conclusion: lnc-NEAT1 may induce osteoporosis viapromoting osteoclaste differentiation and inhibitosteoblastdifferentiation, providing experimental evidence for NEAT1 as a potential therapeutic target for osteoporosis.