LncRNA-COX-2通过TGFβ1/Smad3信号通路参与脊柱结核发病机制的研究

Long non-coding RNA (LncRNA) regulated the expression of protein-coding genes at both the transcriptional and post-transcriptional levels, and they played an important regulatory role in the progression and progression of inflammation and tumors. However, previous studies had rarely focused on LncRNAs of the regulation of the pathogenesis of spinal tuberculosis. Our previous study found that LncRNA-COX-2 was highly expressed in vertebral body lesions of spinal tuberculosis, suggesting that LncRNA-COX-2 was involved in the pathogenesis of spinal tuberculosis. This study intends to use expression profiling microarray method to study high expression of LncRNA-COX-2 in spinal tuberculosis disc and vertebral lesions, and to explore the role of LncRNA-COX-2 in the regulation of spinal tuberculosis. Analysis of LncRNA-COX-2, TGF-β1 , Smad3 and inflammatory factors in rabbit with spinal tuberculosis, in vitro tuberculosis-infected macrophages, and human spinal tuberculosis tissue samples to elucidate the involvement of LncRNA-COX-2 through the TGF-β1/Smad3 signaling pathway. The pathogenesis of tuberculosis and the pathogenesis of spinal tuberculosis are of great significance. This study provided a theoretical basis for the early diagnosis and treatment of spinal tuberculosis targeting LncRNA-COX-2 and drug development for anti-tuberculosis.

长链非编码RNA(LncRNA)在转录及转录后水平调节蛋白编码基因的表达，在炎症和肿瘤的进展和转归过程中起着重要调节作用，但以往的研究很少有关于LncRNA在脊柱结核发病机制中调控作用。我们前期研究发现脊柱结核椎体病灶中LncRNA-COX-2高表达，提示LncRNA-COX-2参与脊柱结核的发病。本课题拟利用表达谱芯片方法进行脊柱结核椎间盘及椎体病变组织中高表达LncRNA-COX-2的功能研究，探索LncRNA-COX-2在脊柱结核发病中的调控作用。通过体内兔脊柱结核、体外结核菌感染巨噬细胞及人体脊柱结核组织标本中的LncRNA-COX-2、TGF-β1、Smad3及炎性因子分析，对阐明LncRNA-COX-2通过TGF-β1/Smad3信号通路参与结核发病、揭示脊柱结核的发病机制有重要意义。本研究为以LncRNA-COX-2为靶点的脊柱结核早期诊断、治疗及抗结核药物研发提供理论基础。

结核病是一类严重危害人类健康的慢性传染性疾病。脊柱结核（STB）是最常见的骨与关节结核，STB的发生与多种细胞和细胞因子有关、多条通路参与其发病，在这个过程中具有调节功能的长链非编码RNA（LncRNAs）、环状RNA（CircRNAs）的表达异常可能在STB的发病机制中发挥重要的调节作用。本研究对STB外周血及椎间盘病灶组织中差异LncRNA、CircRNA及mRNA进行筛选、验证及生物学分析，结果表明STB与退变椎间盘的LncRNA及mRNA的表达量存在着差异。差异表达的LncRNA可能通过与trans-靶蛋白结合的方式调控cis-靶基因表达，进而通过多种信号通路导致人体炎症、免疫功能变化，降低人体对结核分枝杆菌（MTB）的免疫清除能力。LINC01128-201、Lnc-DNALI1-205在STB椎间盘间存在明显的低表达，有成为STB诊断标记物的潜力。STB椎间盘病灶组织中差异表达的circRNA可能与STB的发生和发展密切相关。hsa_circ_0001021、hsa_circ_0043898和hsa_circ_0093669在STB外周血中可作为新型诊断标志物，联合ESR和CRP可提高诊断价值。体外实验表明MTB感染的细菌量与TGF-β1/Smads经典信号通路激活有关。TGF-β1/Smads 通路与STB发病相关，外周血及椎间盘组织TGF-β1、Smad3 mRNA 检测可辅助STB诊断及疗效评价。LBP、S100-A8和S100-A9蛋白可能是STB发病相关的重要表达蛋白，LBP蛋白可能作为STB的诊断标志物。宁夏STB主要以农村、中壮年居多，应加强农民宣教，关注中壮年，注重肺结核防控及STB的早期筛查。STB常见的病理改变为干酪样坏死、多核巨细胞、肉芽肿性炎、结核结节等。布氏杆菌性脊柱炎（BS）的病理改变以肉芽组织增生修复为主，慢性炎症急性发作时泡沫细胞反应和组织细胞学反应明显。STB和BS血常规某些参数、AST、ALT、ALP、ALB存在差异，可能在STB和BS的鉴别诊断、疗效评估等方面具有临床应用价值。本研究对阐明LncRNA、CircRNA等小分子通过TGF-β1/Smad3通路参与结核发病、揭示STB的发病机制有重要意义，为以LncRNA、CircRNA及外周血蛋白分子为靶点的STB诊断标志物、疗效评价及抗结核药物研发提供理论基础。