Effect of LncRNA XIST on osteogenic differentiation in periodontitis rats by regulating the miR-424-5p/SOCS6 axis
Objective:To investigate the effect of long-chain non coding RNA X inactive specific transcript(lncRNA XIST)on osteogenic differentiation in periodontitis rats by regulating the microRNA-424-5p(miR-424-5p)/suppressor of cy-tokine signaling 6(SOCS6)axis.Methods:A rat model of periodontitis was induced by silk thread ligation combined with bacterial injection,and was separated into model group,oe-NC group,oe-XIST group,oe-XIST+NC agomir group,oe-XIST+miR-424-5p-agomir group.An additional 10 rats were selected as the control group.Rat primary PDLSC was isolated,cul-tured,and separated into NC group,tumor necrosis factor(TNF)-α group,oe-NC group,oe-XIST group,oe-XIST+miR-424-5p-NC group,oe-XIST+miR-424-5p-mimics group.Except for the NC group,all the cells in other groups used TNF-α to cul-ture PDLSC to induce periodontitis cell model.ELISA was applied to detect the levels of serum inflammatory factors in rats.HE staining was applied to detect the pathological morphology of rat periodontal tissue.RT-qPCR was applied to detect the expression levels of LncRNA XIST,miR-424-5p,and SOCS6 mRNA in periodontal tissue and PDLSC.CCK-8 was applied to detect cell proliferation.Alizarin red staining was applied to detect the osteogenic differentiation ability of cells.The reagent kit was applied to measure ALP activity in cells.Western blot was applied to detect the expression of SOCS6 and osteogenic differentiation related proteins in cells.Dual luciferase reporter gene experiment was applied to verify the targeting relation-ship between LncRNA XIST,miR-424-5p,and SOCS6.Results:In rats experiment of periodontitis,overexpression of LncRNA XIST can improve the pathological damage of rat periodontal tissues,the levels of TNF-α,interleukin(IL)-1β,IL-6 and the expression of miR-424-5p in serum of rats were decreased,and the expression of LncRNA XIST and SOCS6 mRNA was in-creased(P<0.05).In the PDLSC experiment,overexpression of LncRNA XIST can increase cell survival rate,calcium depo-sition,ALP activity,the expression of LncRNA XIST and SOCS6 mRNA,and the expression of SOCS6,RUNX2 and OCN pro-teins induced by TNF-α,and decrease the levels of TNF-α,IL-1β,IL-6 and miR-424-5p in cells(P<0.05).However,the co-expression of LncRNA XIST and miR-424-5p further aggravated the periodontal tissues damage of rats,decreased the survival rate,calcium deposition,ALP activity,SOCS6 mRNA expression and SOCS6,RUNX2 and OCN protein expression of PDLSC cells,and increased the levels of TNF-α,IL-1β,IL-6 and miR-424-5p expression in cells(P<0.05).Conclusion:LncRNA XIST may promote osteogenic differentiation in periodontitis rats by regulating the miR-424-5p/SOCS6 signaling axis.
PeriodontitisOsteogenic differentiationLong-chain non coding RNA X-inactive specific transcriptMi-cro RNA-424-5pSuppressor of cytokine signaling 6
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