静态牵张力通过下调linc01135/mir-106a-5p抑制pPDLSCs成骨分化

Static mechanical stress inhibits osteogenic differentiation of pPDLSCs through downregulating linc01135/mir-106a-5p

目的:研究静态牵张力通过长链非编码RNA linc01135调节炎症来源牙周膜干细胞成骨分化的机制。方法:使用RT-qPCR法检测加载静态牵张力后牙周膜干细胞中linc01135的表达水平。使用慢病毒转染的方法调节linc01135和mir-106a-5p的表达水平,RT-qPCR法检测成骨相关基因的表达水平以及使用茜素红染色实验检测细胞的成骨分化能力。AGO2-rip和双荧光素酶报告基因实验验证linc01135与mir-106a-5p的结合互作。结果:炎症来源牙周膜干细胞加力后linc01135表达水平降低,RT-qPCR和茜素红染色结果表明linc01135过表达促进细胞的成骨分化能力,mir-106a-5p抑制细胞的成骨分化能力,AGO2-rip和双荧光素酶报告实验表明linc01135可以与mir-106a-5p结合互作。结论:炎症条件下静态牵张力通过下调linc01135,降低了linc01135对mir-106a-5p的内源性结合作用,进而抑制细胞的成骨分化。

Objective:To investigate the mechanism that static mechanical strain(SMS)affecting osteogenic differentiation of periodontal ligament stem cells from periodontitis(pPDLSCs)via long non-coding RNA linc01135.Methods:RT-qPCR was performed to detect the expression level of linc01135 in periodontal ligament stem cells from health(hPDLSCs)and pPDLSCs.The expression levels of linc01135 and mir-106 a-5 p were regulated by lentiviruses.RT-qPCR was performed to detect the expression level of osteogenic genes and alizarin red staining assay was performed to detect the osteogenic differentiation ability.AGO2-rip and dual luciferase reporter gene assays were performed to verify the interactions of linc01135 and mir-106 a-5 p.Results:The expression level of linc01135 in pPDLSCs was reduced after loading with SMS.Results of RT-qPCR and alizarin red staining showed that linc01135 overexpression promoted the osteogenic differentiation ability however mir-106 a-5 p inhibited the osteogenic differentiation ability.AGO2-rip and dual luciferase reporter assay showed that linc01135 could interact with mir-106 a-5 p.Conclusion:SMS reduces the endogenous interaction effect of linc01135 on mir-106 a-5 p by downregulating linc01135,which inhibited osteogenic differentiation of pPDLSCs.