人退变腰椎间盘组织中p53与IL-1β、IL-6表达的实验研究

发布时间：2018-04-16 23:40

本文选题：椎间盘退变 + 细胞老化　；参考：《东南大学》2015年硕士论文

【摘要】：目的：研究表明椎间盘退变与椎间盘细胞老化以及老化细胞分泌的炎性因子密切相关,本实验通过检测p53和白细胞介素-1β(IL-1β)、白细胞介素-6(IL-6)在退变与正常的腰椎间盘髓核组织中的表达,比较两组的表达水平,分析p53与IL-1β、IL-6表达的相关性,以及椎间盘退变等级与p53、IL-1β、IL-6表达的相关性。探讨p53、IL-1β、IL-6在椎间盘退变中的作用及意义。方法：收集40例腰椎间盘突出症患者的椎间盘髓核组织,其中男22例,女18例,突出部位L4/5,术后病理证实为退变椎间盘髓核组织,并纳入退变组。另取20例腰椎爆裂型骨折手术患者受累椎间盘髓核组织,其中男14例,女6例,受累部位L4/5,术后病理证实为正常椎间盘髓核组织,并纳入正常组。将髓核根据Thompson分级分为五级。应用免疫组织化学方法检测髓核标本中p53与IL-1β、IL-6的表达。分别记录三者的细胞染色阳性率并进行分级,采用X2检验方法(chi-square test)对三者在退变组与正常组的细胞染色阳性率进行比较,同时采用spearman等级相关分析方法分析椎间盘退变等级与p53、IL-1β、IL-6表达的相关性。应用Western-Blot技术检测髓核标本中p53与IL-1β、IL-6的表达,两组之间采用独立样本t检验进行比较,同时采用spearman等级相关分析方法分析p53分别与IL-1β、IL-6表达的相关性。结果：1、免疫组织化学方法检测结果：退变组中40例髓核P53、IL-1β、IL-6染色阳性率分别为80.0%(32/40)、87.5%(35/40)和72.5%(29/40),正常组中p53、IL-1β、IL-6染色阳性率分别为10%(2/20)、20%(4/20)和15%(3/20),p53、IL-1β、IL-6分别在两组之间的表达差异有显著意义(p0.001)。椎间盘退变等级分别与p53、IL-1β、IL-6量的表达正相关(相关系数分别为rs=0.616,p0.001,显著相关；rs=0.673,p0.001,显著相关；rs=0.654,p0.001,显著相关)。2、Western Blot结果表明退变组中p53与IL-1β、IL-6蛋白表达量相对于正常组均有明显增加,两组中p53与IL-1β、IL-6灰度比值有统计学差异(P0.05)。p53分别与IL-1β、IL-6的表达呈正相关,p53与IL-1β与的相关系数为rs=0.466,p=0.003,中度相关；p53与IL-6的相关系数为rs=0.447,p=0.004,中度相关。IL-1p与IL-6的表达无相关性,其相关系数分别为rs=0.113,p=0.512。结论：1、p53与IL-1β、IL-6在椎间盘退变过程中表达增高,三者与椎间盘退变过程关系密切。2、p53、IL-1β、IL-6与椎间盘退变等级之间存在正相关,三者是导致椎间盘退变的重要原因。3、p53与IL-1β、IL-6之间存在正相关性,p53在椎间盘退变过程中发挥作用与IL-1β、IL-6有关。
[Abstract]:Objective: to study that disc degeneration is closely related to aging of intervertebral disc cells and inflammatory factors secreted by aging cells of intervertebral disc.In this study, the expression of p53, interleukin-1 尾, interleukin-6 (IL-6) in the nucleus pulposus of degenerative and normal lumbar intervertebral disc was detected, and the expression levels of p53 and IL-1 尾 -IL-6 were compared between the two groups, and the correlation between the expression of p53 and IL-1 尾 -IL-6 was analyzed.And the correlation between the grade of degeneration of intervertebral disc and the expression of IL-1 尾-6.To investigate the role and significance of P 53 and IL 1 尾 IL 6 in the degeneration of intervertebral disc.Methods: the nucleus pulposus tissues of 40 patients with lumbar disc herniation were collected, including 22 males and 18 females, and L4 / 5 of the protrusion site. The degenerative nucleus pulposus tissue was confirmed by postoperative pathology and included in the degenerative group.Another 20 patients with lumbar burst fracture were involved in the nucleus pulposus of intervertebral disc, including 14 males and 6 females, and the involved site was L4 / 5. The pathological results showed that the tissue was normal and included in the normal group.The nucleus pulposus was divided into five grades according to Thompson classification.The expression of p53 and IL-1 尾-sil-6 in nucleus pulposus was detected by immunohistochemical method.The positive rate of cell staining was recorded and graded. The positive rate of cell staining was compared between the three groups in the degenerative group and the normal group by using X2 test.At the same time, spearman grade correlation analysis was used to analyze the correlation between the grade of disc degeneration and the expression of IL-1 尾-6 in p53.The expression of p53 and IL-1 尾 -IL-6 in nucleus pulposus was detected by Western-Blot technique, and the correlation between p53 and IL-1 尾 -IL-6 was analyzed by spearman rank correlation analysis.Results: in the degenerative group, the positive rates of P53P IL-1 尾 and IL-6 were 80.032 / 40 / 87.5g / 35 / 40 and 72.52.5 / 29 / 40, respectively. The positive rates of p53P / IL-1 尾 / IL-6 staining in the normal group were 1022 / 20 / 20 / 420) and 1520 / 30 / 535 / 420, respectively, and there were significant differences between the two groups.The intervertebral disc degeneration grade was positively correlated with the expression of IL-1 尾 -IL-6 (r = 0.616, P 0.001, r = 0.673, P 0.001, P 0.001, P = 0.654, P 0.001, respectively). The results of Western Blot showed that the expression of p53 and IL-1 尾 IL-6 protein in the degenerative group was significantly higher than that in the normal group, and the expression of p53 and IL-1 尾 IL-6 protein in the degenerative group was significantly higher than that in the normal group.There was a significant difference between p53 and IL-1 尾 -IL-6 in gray scale ratio. There was a positive correlation between p53 and IL-1 尾 -IL-6. The correlation coefficient between p53 and IL-1 尾 was 0.466p0.003, the correlation coefficient between p53 and IL-6 was 0.447p0.004, and there was no correlation between the expression of IL-6 and IL-1p, and the correlation coefficient was 0.113p0.512.Conclusion the expression of p53 and IL-1 尾 -IL-6 in intervertebral disc degeneration is increased, and there is a positive correlation between p53 and IL-1 尾 -IL-6 and the grade of intervertebral disc degeneration.There is a positive correlation between p53 and IL-1 尾 -IL-6. The role of p53 in the process of intervertebral disc degeneration is related to IL-1 尾 -IL-6.
【学位授予单位】：东南大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R681.53

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