HOTAIR/miR-21/PTEN的ceRNA网络调节髓核细胞自噬在椎间盘退变中的作用机制

Intervertebral disc degeneration (IDD) is an important cause of low back pain and its precise aetiology is not fully understood. There is a growing body of evidence that autophagy disfunction is extensively involved in the development of IDD. LncRNA-miRNA-mRNA function as ceRNA network，a new pattern of gene expression，widely found in various disease, but the detail function and mechanism of ceRNA network in IDD has not been reported yet. Recently, we detected the expression of miRNAs in human degeneratve and normal NP specimens by the miRNA microarray. MiR-21, one of these miRNAs, is highly upregulated in IDD. Bioinformatics analysis had found that both HOTAIR and PTEN can bind with miR-21. Next, we found that HOTAIR expression was downregulated in nucleus pulposus (NP) cells and was gradually downregulated with aggravation of disc degeneration degree. Moreover, overexpression of HOTAIR can promoted the degradation of type II collagen (Col II) and aggrecan in vitro. We exploited the relationship of HOTAIR and extracellular matrix (ECM) may be attributed to HOTAIR/miR-21/PTEN ceRNA network in IDD. Furthermore, we clarified the detail molecular mechanisms of the relationship of HOTAIR/miR-21/PTEN and cell autophagy or ECM in vivo and in vitro. In generally, we want to confirm the hypothesis that “HOTAIR/miR-21/PTEN function as ceRNA network to regulate NP cells autophagy, thereby controlling ECM cleavage the in IDD”, which may represent a potential therapeutic target for the treatment of IDD.

椎间盘退变(IDD)是导致腰痛的主要原因，发病机制不明，仍无有效治疗方案。髓核细胞(NP)自噬影响胞外基质代谢，诱发椎间盘稳态失衡，将影响IDD过程。课题前期发现miR-21在NP差异表达，并影响NP自噬和胞外基质代谢；HOTAIR调节NP胞外机制合成；生物信息预测PTEN和HOTAIR与miR-21有共同结合位点，然而HOTAIR/miR-21/PTEN是否通过构成ceRNA网络影响NP自噬进而调节胞外基质代谢机制不明。本项目拟采用分子生物、免疫荧光、电镜技术检测人体髓核组织HOTAIR、miR-21和PTEN表达与NP自噬和IDD的相关性；进一步采用人体细胞和动物模型，模拟IDD，采用基因工程技术干扰关键基因表达，利用同步辐射、荧光素酶实验等技术阐明HOTAIR/miR-21/PTEN的ceRNA网络参与调节NP自噬及胞外基质代谢的分子机制及在IDD中的作用，为IDD的治疗提供新靶点。

椎间盘退变(IDD)是导致腰痛的主要原因，发病机制不明，仍无有效治疗方案。髓核细胞(NP)自噬影响胞外基质代谢，诱发椎间盘稳态失衡，将影响IDD过程。课题前期发现miR-21在NP差异表达，并影响NP自噬和胞外基质代谢；HOTAIR调节NP胞外基质合成；生物信息预测PTEN和HOTAIR与miR-21有共同结合位点，然而HOTAIR/miR-21/PTEN是否通过构成ceRNA网络影响NP自噬进而调节胞外基质代谢机制不明。本项目拟采用分子生物、免疫荧光、电镜技术检测人体髓核组织HOTAIR、miR-21和PTEN表达与NP自噬和IDD的相关性；进一步采用人体细胞和动物模型，模拟IDD，采用基因工程技术干扰关键基因表达，利用同步辐射、荧光素酶实验等技术阐明HOTAIR/miR-21/PTEN的ceRNA网络参与调节NP自噬及胞外基质代谢的分子机制及在IDD中的作用，为IDD的治疗提供新靶点。