【作者】 谭清实；

【导师】 徐宝山； 夏群；

【作者基本信息】 天津医科大学 ， 外科学， 2010， 硕士

【摘要】 目的：探讨椎间盘源性腰痛的发病机制及与椎间盘突出、脱出及对照组的异同。方法：收集71例因椎间盘退行性疾病行手术治疗的病人椎间盘标本,临床诊断包括椎间盘源性腰痛10例,椎间盘突出症52例(椎间盘突出30例,髓核脱出或游离22例)。另收集9例因腰椎骨折行前路手术切除的伤椎远侧相对正常椎间盘(MRI无明显退变)为对照组。取髓核组织用生理盐水清洗,即刻-80℃冰箱保存。标本分成3部分行以下实验：(1)HE染色、光学显微镜下观察细胞形态、数量及分布,细胞基质退变程度、炎细胞浸润程度和小血管增生情况。(2)应用免疫组化SP法对同一部位连续切片的CD68巨噬细胞和CD45RO阳性T细胞染色,观察CD68巨噬细胞和CD45RO阳性T细胞的表达。(3)髓核组织匀浆后取上清液,用酶联免疫吸附试验(Elisa)测定细胞因子(IL-1、IL-6、TNF-α)的含量。用SPSS13.0统计软件分析。结果：(1)HE染色对照组髓核细胞形态大小一致,分布均匀,未见明显细胞基质退变及炎性细胞浸润。其他各组均见髓核细胞空泡样变、形态大小不一、分布不均。椎间盘脱出组、椎间盘突出组见细胞基质退变,髓核组织边缘可见大量炎性细胞浸润、局灶性小血管增生。椎间盘源性腰痛组细胞基质严重退变,髓核组织边缘可见散在炎性细胞浸润,未见明显血管增生。(2)免疫组化病变各组中均有CD68巨噬细胞表达,阳性率(阳性数/总数)：脱出组(14/22)>椎间盘源性腰痛组(4/10)>突出组(8/30),对照组CD68巨噬细胞表达阴性,卡方检验四组之间有统计学意义(P<0.05),经两两比较：脱出组与突出组、脱出组与对照组差异有统计学意义；CD45RO阳性T细胞均出现在CD68巨噬细胞同一部位的连续切片上,CD45RO阳性T细胞阳性率(阳性数/总数)：脱出组(13/22)>突出组(4/30)，在间盘源性腰痛组和对照组中未见阳性表达,卡方检验四组之间有统计学意义(P<0.05),经两两比较：脱出组与椎间盘源性腰痛组、脱出组与突出组、脱出组与对照组差异有统计学意义。(3)Elisa检测IL-1含量均值(pg/mL)自大至小依次为：椎间盘源性腰痛组12.32>椎间盘脱出组11.84>椎间盘突出组7.89>对照组5.16；秩和检验四组之间有统计学意义(P<0.05),经两两比较：间盘源性腰痛组与突出组、间盘源性疼痛组与对照组、脱出组与突出组、脱出组与对照组差异有统计学意义。IL-6含量均值自大至小依次为：椎间盘脱出组128.02>椎间盘突出组17.59>椎间盘源性腰痛组18.6>对照组7.03；秩和检验四组之间有统计学意义(P<0.05),经两两比较：脱出组与突出组、脱出组与对照组、突出组与对照组、间盘源性腰痛组与对照组差异有统计学意义。TNF-α含量均值(pg/ML)自大至小依次为：椎间盘脱出组15.61>椎间盘突出组13.74>椎间盘源性腰痛组11.54>对照组5.40；秩和检验四组之间有统计学意义(P<0.05),经两两比较：各组与对照组差异均有统计学意义。结论：(1)椎间盘源性腰痛髓核细胞基质退变严重,髓核组织中有CD68巨噬细胞和细胞因子IL-1、IL-6、TNF-α,说明炎性反应和细胞因子是其发病机制的重要因素。(2)椎间盘脱出、突出的髓核组织边缘有大量巨噬细胞、CD45RO免疫T细胞和小血管增生；而椎间盘源性腰痛髓核组织边缘炎性细胞浸润较少,无明显的小血管增生和CD45RO免疫T细胞,提示椎间盘源性腰痛尚有细胞免疫以外的发病机制。更多还原

【Abstract】 Objective:To study the pathogenesis of degenerative disc disease such as discogenic low back pain and the difference from disc herniation, prolapse and control group.Method:71 cases disc specimens with degenerative disc disease were collected during operation, including 10 cases in discogenic low back pain,52 cases Disc herniation(30 cases in disc herniation,22 cases in disc prolapse), And nine cases of spinal fractures were collected during anterior approach as normal control group (MRI shows no degeneration), The nucleus pulposus were got during surgery, washed with saline and were immediately placed in-80℃refrigerator after surgery. (1)HE staining, observe the morphology, size and distribution of the nucleus pulposus cells, observe the degree of degeneration of extracellular matrix, inflammatory cells infiltration and small vessel hyperplasia. (2) SP immunohistochemical was used to stain CD68 positive macrophages and CD45RO T cells. (3)Break down the nucleus pulposus tissues, get the supernate, determine the the content of the cytokines of IL-1, IL-6, TNF-a by Elisa. Analyzed with SPSS 13.0 Statistics software.Result:(1) HE staining:In the control group, the nucleus pulposus cells are in the same shape and size, and are well distributed, no obvious cell matrix degeneration can be seen. All of the other groups can see that nucleus pulposus cells became vascuolated, different in size and distribution, there are inflammatory cells and focal small vessels proliferation on the edge of disc prolapse group, herniation group, In discogenic low back pain group the cell matrix became degenerate severely, there are inflammatory cells but there is no significant angiogenesis;(2)Immunohistochemistry CD68-positive macrophages exist in each disease group, positive rate(positive cases/all cases):prolapse group(14/22)>discogenic low back pain group(4/10)> herniation group(8/30)>, there is no positive cells can be seen in control group, Chi-square test reveals that there is significant difference between the four groups (P <0.05), by comparison:There is significant difference between prolapse group and herniation group, as well as prolapse group and control group. positive rate of CD45RO T cells (positive cases/all cases):prolapse group(13/22)>herniation group(4/30), there is no positive cells can be seen in discogenic low back pain group and control group, Chi-square test reveals that there is significant difference between the four groups(P<0.05), by comparison:There is significant difference between prolapse group and discogenic low back pain group, prolapse group and herniation group, prolapse group and control group. (3) Elisa test Mean Content of IL-1(pg/ML) form high to low:discogenic low back pain group12.32>prolapse group11.84> herniation group7.89>control group5.16, Rank sum test demonstrate there is significant difference between the four groups(P<0.05), by comparison:There is significant difference between discogenic low back pain group and herniation group, discogenic low back pain group and control group, prolapse group and herniation group, prolapse group and control group. Mean Content of IL-6(pg/ML) form high to low:prolapse groupl28.02>herniation group17.59>discogenic low back pain group18.6>control group7.03; Rank sum test demonstrate there is significant difference between the four groups(P<0.05), by comparison:There is significant difference between prolapse group and herniation group, prolapse group and control group, discogenic low back pain group and control group. Mean Content of TNF-a(pg/ML) form high to low:prolapse group 15.61>herniation group13.74> discogenic low back pain group 11.54>control group5.40, Rank sum test demonstrate there is significant difference between the four groups (P<0.05), by comparison:There is significant difference between each group and control group.Conclusion:(1)Cell matrix of discogenic low back pain degenerates seriously, there are CD68- positive macrophages and cytokines of IL-1, IL-6, TNF-a in the nucleus pulposus, which suggest inflammatory response and cytokines are important pathoge-nesis. (2) There are macrophages, CD45RO T cells and focal small vessels proliferatio- n on the edge of prolapse group, herniation group; on the edge of low back pain group inflammatory cells are not so many, and there is no small vessels proliferation and CD45RO T cells, which suggest the pathogenesis of discogenic low back pain dose not associated with cellular immunity.更多还原