miR-455-3p靶向HIPK2调控高糖环境下成骨细胞的增殖和凋亡

文题释义： 糖尿病性骨病：糖尿病可导致骨代谢发生改变，表现为骨形成缺陷、成骨细胞数量减少、骨基质形成不足。高糖环境下以成骨细胞功能改变最为显著，高糖诱导成骨细胞是研究糖尿病性骨病的常用细胞模型。 成骨作用：指成骨细胞移至将要合成骨组织的部位，分泌、合成骨胶原及骨蛋白纤维，将钙、磷吸收到纤维孔隙中进行沉淀结晶，形成骨组织的过程。 背景：以往研究表明多种miRNA在骨形成中发挥作用，miR-335-5p可保护成骨细胞免受氧化应激，对枸橼酸铁铵诱导的成骨细胞具有保护作用，但miR-335-5p对高糖环境下成骨细胞增殖和凋亡的影响尚未可知。 目的：探讨miR-455-3p靶向HIPK2对高糖环境下成骨细胞增殖和凋亡的影响。 方法：双荧光素酶报告基因分析法验证miR-455-3p对HIPK2的靶向作用。体外用高糖诱导MC3T3-E1细胞，分为空白组、高糖组、高糖+miR-control组、高糖+miR-455-3p组、高糖+si-control组、高糖+si-HIPK2组、高糖+miR-455-3p+pcDNA组和高糖+miR-455-3p+pcDNA-HIPK2组。qRT-PCR检测miR-455-3p和HIPK2 mRNA的表达，MTT法检测细胞存活率，流式细胞检测细胞凋亡，Western blot检测HIPK2、p-STAT3和STAT3蛋白的表达。 结果与结论：①HIPK2是miR-455-3p的靶基因，miR-455-3p可负性调控HIPK2的表达；②高糖处理可抑制miR-455-3p的表达，促进HIPK2的表达；③过表达miR-455-3p或抑制HIPK2表达均可促进高糖条件下MC3T3-E1细胞的存活和抑制凋亡；④过表达HIPK2可部分逆转miR-455-3p对高糖条件下成骨细胞的存活促进和凋亡抑制作用；⑤miR-455-3p通过调控HIPK2抑制成骨细胞中p-STAT3的表达；⑥结果表明，miR-455-3p通过靶向下调HIPK2抑制高糖诱导的成骨细胞凋亡，促进成骨细胞增殖，这可能与抑制STAT3信号通路有关。 ORCID: 0000-0001-7129-8455(匡嘉兵) 中国组织工程研究杂志出版内容重点：组织构建；骨细胞；软骨细胞；细胞培养；成纤维细胞；血管内皮细胞；骨质疏松；组织工程

Effect of miR-455-3p targeting HIPK2 on proliferation and apoptosis of osteoblasts induced by high glucose

BACKGROUND:Previous studies have shown that various miRNAs play a role in bone formation.miR-335-5p can protect osteoblasts from oxidative stress and protect osteoblasts under induction with ferric ammonium citrate,but the effect of miR-335-5p on osteoblast proliferation and apoptosis in high glucose environments is unknown.OBJECTIVE:To investigate the effect of miR-455-3p targeting HIPK2 on proliferation and apoptosis of osteoblasts induced by high glucose.METHODS:Dual luciferase reporter assay was used to verify the targeting of miR-455-3p to HIPK2.MC3T3-E1 cells were induced by high glucose in vitro,and MC3T3-E1 cells were treated as follows:blank group,high glucose group,high glucose+miR-control group,high glucose+miR-455-3p group,high sugar+si-control group,high sugar+si-HIPK2 group,high glucose+miR-455-3p+pcDNA group and high glucose+miR-455-3p+pcDNA-HIPK2 group.The expression of miR-455-3p and HIPK2 mRNA was detected by qRT-PCR,cell viability was detected by MTT assay,apoptosis was detected by flow cytometry,and the expression of HIPK2,p-STAT3 and STAT3 protein was detected by western blot.RESULTS AND CONCLUSION:HIPK2 was a target gene of miR-455-3p,and miR-455-3p negatively regulated the expression of HIPK2.High glucose treatment inhibited the expression of miR-455-3p and promoted the expression of HIPK2.The over-expression of miR-455-3p or the inhibition of HIPK2 promoted MC3T3-E1 survival and inhibit cell apoptosis after high glucose treatment.The over-expression of HIPK2 partially reversed the survival promotion and apoptosis inhibition of miR-455-3p on osteoblasts induced by high glucose.miR-455-3p inhibited the expression of p-STAT3 in osteoblasts by regulating HIPK2.To conclude,miR-455-3p inhibits the apoptosis and promotes the proliferation of osteoblasts induced by high glucose via down-regulating HIPK2,which may be related to the inhibition of STAT3 signaling pathway.