微小RNA-210/缺氧诱导因子1α信号轴与椎间盘退变程度的相关性分析

Correlation analysis of mircoRNA-210/hypoxia inducible factor-1α signaling axis and intervertebral disc degeneration

中文关键词:  腰椎间盘突出症  椎间盘退变  缺氧  缺氧诱导因子1α  微小RNA-210

英文关键词:Lumbar disc herniation  Intervertebral disc degeneration  Hypoxia  Hypoxia inducible factor-1α  MircoRNA-210

中文摘要:

  【摘要】 目的：探讨微小RNA-210（mircoRNA-210，miR-210）/缺氧诱导因子1α（hypoxia inducible factor-1α，HIF-1α）信号轴与椎间盘退变程度的相关性。方法：选取2024年1月~12月在苏州市中医医院骨伤科行手术治疗切除髓核组织的45例腰椎间盘突出症（lumbar disc herniation，LDH）患者，男22例，女23例，年龄20~75岁[59（45，67）岁]；病程1~240个月[36（9，120）个月]。收集患者术前影像学资料及术中切除的髓核组织样本。在术前矢状位MRI T2加权像上采用Pfirrmann分级系统对手术椎间盘的退变程度进行分级；对髓核组织固定切片后行苏木精-伊红（hematoxylin-eosin，HE）染色观察椎间盘病理改变并行组织学评分；应用定量反转录聚合酶链式反应（quantitative reverse transcription polymerase chain reaction，qRT-PCR）检测髓核组织中miR-210-3p及HIF-1α、甘油-3-磷酸脱氢酶1样蛋白（glycerol-3-phosphate dehydrogenase 1-like，GPD1L）mRNA表达水平；应用免疫组化染色检测髓核组织中HIF-1α、GPD1L蛋白表达水平。分析年龄、病程、组织学评分与Pfirrmann分级，miR-210-3p、HIF-1α、GPD1L表达水平与椎间盘退变程度及miR-210-3p、HIF-1α、GPD1L之间的相关性。结果：45例患者手术椎间盘Pfirrmann分级包括Ⅱ级4例，Ⅲ级14例，Ⅳ级19例，Ⅴ级8例。Pfirrmann分级与年龄及病程正相关（rs=0.763，P<0.001；rs=0.715，P<0.001）。髓核组织病理组织学评分显示，Pfirrmann Ⅱ级组3.00±0.54分，Pfirrmann Ⅲ级组5.14±0.60分，Pfirrmann Ⅳ级组8.58±0.74分，Pfirrmann Ⅴ级组9.54±0.75分，组间差异具有统计学意义（F=136.540，P＜0.001），组织学评分与Pfirrmann分级正相关（rs=0.889，P<0.001）。不同Pfirrmann分级髓核组织miR-210-3p、HIF-1α mRNA、HIF-1α蛋白、GPD1L mRNA、GPD1L蛋白表达水平存在显著性差异（P＜0.05），髓核组织miR-210-3p表达与Pfirrmann分级及组织学评分正相关（rs=0.890，P<0.001；rs=0.832，P＜0.001），HIF-1α mRNA和蛋白表达与Pfirrmann分级及组织学评分均正相关（rs=0.874，P<0.001；rs=0.821，P<0.001和rs=0.868，P<0.001；rs=0.820，P<0.001），GPD1L mRNA和蛋白表达与Pfirrmann分级及组织学评分均负相关（rs=－0.807，P<0.001；rs=－0.722，P<0.001和rs=－0.793，P<0.001；rs=－0.718，P<0.001）。髓核组织miRNA-210-3p与HIF-1α mRNA表达正相关（rs=0.839，P<0.001），与GPD1L mRNA表达负相关（rs=－0.776，P<0.001）；HIF-1α与GPD1L mRNA及蛋白表达均负相关（rs=－0.813，P<0.001和rs=－0.754，P<0.001）。结论：miR-210/HIF-1α信号轴与椎间盘退变程度相关联，miR-210-3p、GPD1L和HIF-1α互相影响，共同参与椎间盘退变的分子机制。

英文摘要:

  【Abstract】 Objectives: To investigate the correlation between the microRNA-210(miR-210)/hypoxia inducible factor-1α(HIF-1α) signaling axis and intervertebral disc degeneration. Methods: From January to December 2024, 45 patients with lumbar disc herniation(LDH) who underwent surgical treatment for nucleus pulposus removal in the Department of Orthopedics and Traumatology, Suzhou Traditional Chinese Medicine Hospital were selected. There were 22 males and 23 females, aged 20 to 75 years[59(45, 67) years]. The disease duration was 1 to 240 months[36(9, 120) months]. Preoperative imaging data and the removed nucleus pulposus samples during operation were collected. The degeneration degree of the surgical intervertebral discs was graded using the Pfirrmann classification system on the sagittal MRI T2-weighted images; Hematoxylin-eosin(HE) staining was performed on the fixed sections of the nucleus pulposus to observe the pathological changes of the intervertebral discs and conduct histological scoring; Quantitative reverse transcription polymerase chain reaction(qRT-PCR) was used to detect the mRNA expression levels of miR-210-3p, HIF-1α, and glycerol-3-phosphate dehydrogenase 1-like(GPD1L) in the nucleus pulposus tissue; Immunohistochemical staining was used to detect the protein expression levels of HIF-1α and GPD1L in the nucleus pulposus tissue. The correlations between age, disease duration, histological scores and Pfirrmann grading, as well as the correlations between miR-210-3p, HIF-1α, GPD1L expression levels and intervertebral disc degeneration degree and miR-210-3p, HIF-1α, GPD1L were analyzed. Results: Pfirrmann grades of the surgical intervertebral discs of the 45 patients were: 4 cases of grade Ⅱ, 14 cases of grade Ⅲ, 19 cases of grade Ⅳ, and 8 cases of grade Ⅴ. Pfirrmann grading was positively correlated with age and disease duration(rs=0.763, P<0.001; rs=0.715, P<0.001). The pathological histological scores of nucleus pulposus were: 3.00±0.54 in Pfirrmann grade Ⅱ, 5.14±0.60 in grade Ⅲ, 8.58±0.74 in grade Ⅳ, and 9.54±0.75 in grade Ⅴ groups. The differences between groups were statistically significant(F=136.540, P<0.001), and the histological scores were positively correlated with Pfirrmann grade(rs=0.889, P<0.001). The expression levels of miR-210-3p, HIF-1α mRNA, HIF-1α protein, GPD1L mRNA, and GPD1L protein in nucleus pulposus tissue differed significantly among different Pfirrmann grades(P<0.05). The expression of miR-210-3p in nucleus pulposus tissue was positively correlated with Pfirrmann grade and histological score(rs=0.890, P<0.001; rs=0.832, P<0.001). The mRNA and protein expression of HIF-1α was positively correlated with Pfirrmann grade and histological score(rs=0.874, P<0.001; rs=0.821, P<0.001, and rs=0.868, P<0.001; rs=0.820, P<0.001). The mRNA and protein expression of GPD1L was negatively correlated with Pfirrmann grade and histological score(rs=-0.807, P<0.001; rs=-0.722, P<0.001, and rs= -0.793, P<0.001; rs=-0.718, P<0.001). The expression of miR-210-3p was positively correlated with HIF-1α mRNA expression(rs=0.839, P<0.001), and negatively correlated with GPD1L mRNA expression(rs=-0.776, P<0.001). HIF-1α expression was negatively correlated with both mRNA and protein expression of GPD1L(rs= -0.813, P<0.001, and rs=-0.754, P<0.001). Conclusions: The miR-210/HIF-1α signaling axis is associated with the degree of intervertebral disc degeneration, miR-210-3p, GPD1L and HIF-1α interact with each other and jointly participate in intervertebral disc degeneration.