低氧诱导因子在腰椎间盘突出中的病理机制研究

Role of HIF in the pathology mechanism of the herniation of lumbar intervertebral disc

目的研究低氧诱导因子(HIF)在腰椎间盘突出中的病理机制。方法采用磁共振图像系统对腰椎间盘突出症进行分级诊断并将收集的样本分为腰椎间盘突出轻度、中度、重度组和对照组;采用免疫组织化学方法对HIF在腰椎间盘突出病人中的表达进行检测、real-time PCR方法检测其mRNA水平上的定量表达、Western blotting方法半定量分析HIF的蛋白表达、TUNEL法检测髓核细胞凋亡率;用Excel软件分析HIF在腰椎间盘突出中的表达与髓核细胞的凋亡的相关性。结果 HIF在髓核细胞的细胞核内积聚,其在腰椎间盘突出组的表达明显高于对照组,且在腰椎盘突出轻度、中度和重度组间,HIF的表达逐步增加(P<0.05)。在腰椎间盘突出组59个样本中,HIF的表达结果显示有53个呈弱阳性、阳性或强阳性。定量real-time PCR检测中,HIF在腰椎间盘突出组和对照组中的表达分别为(0.484±0.079)和(0.241±0.083),P<0.05,差异有统计学意义。HIF在患病轻度组的表达量高于对照组,中度组高于轻度组,重度组高于中度组,P值分别为0.021、0.025、和0.014,均差异有统计学意义。Western blotting方法半定量HIF表达呈现与定量real-time PCR检测分析相一致的结果。髓核细胞的凋亡率在腰椎间盘突出组中(51.0±7.5)%明显高于对照组(14.4±6.2)%,差异有统计学意义(P<0.05);重度组的细胞凋亡率达(75.2±8.4)%,高于中度组(P<0.05);中度组细胞凋亡率高于轻度组,差异有统计学意义(P<0.05)。HIF的表达与腰椎间盘突出髓核细胞的凋亡呈正相关,差异有统计学意义(P<0.05)。结论 HIF可能参与并作用腰椎间盘突出症的病理机制,其表达水平明显增加,并与腰椎间盘突出髓核细胞的凋亡呈现正相关。

Objective To evaluate the role of HIF expression in the pathology mechanism of the herniation of lumbar intervertebral disc.Methods Collected samples were distributed to lumbar disc herniation group （subgroup：mild,moderate and severe groups） and control group by MRI;Immunohistochemistry staining,quantitative real-time PCR and western blotting were utilized to test the expression of HIF in patients with lumbar disc herniation,and TUNEL assay was used to test cell death of nucleus pulposus cells with lumbar disc herniation disease.Correlation analysis was performed by Excel micro software.Results HIF was accumulated on the nucleus pulposus,and its expression in herniation patient group was significantly higher than that in control group （P〈0.05）;HIF expression in moderate group was higher than mild group and less than severe group （P〈0.05）.53 samples out of 59 samples derived from herniation group were evaluated as HIF weak positive,positive,or strong positive results.In quantitative real-time PCR assay,the value of HIF level in herniation group was （0.484±0.079）,which was higher than （0.241±0.083） in control group （P〈0.05）.HIF expression in mild group was higher than control group and less than moderate group;moderate group was less than severe group.Their Pvalues were 0.021,0.025 and 0.014,respectively.The result from western blotting test showed consistent result with quantitative real-time PCR assay.Cell death rate of （51.0±7.5）% in herniation group was significantly higher than （14.4±6.2）% in control group （P〈0.05）.Cell death rate of moderate group was lower than severe group and higher than mild group （P〈0.05）.Expression of HIF in patients with lumbar disc herniation had a positive correlation with cell death of nucleus pulposus cells （P〈0.05）.Conclusions The expression of HIF was increased,and HIF had a positive correlation with apoptosis of nucleus pulposus cells in lumbar disc herniation disease.Therefore,HIF might be involved in the pathology mechanism of lumbar disc herniation.