CD147在神经病理性疼痛模型大鼠脊髓及背根神经节内的表达变化

发布时间：2018-04-17 07:59

本文选题：CD147 + 神经病理性疼痛　；参考：《第四军医大学》2010年硕士论文

【摘要】： 神经病理性疼痛(Neuropathic pain)是指外周或中枢神经系统结构损害、功能障碍或者疾病所致的疼痛,它给患者带来极大的痛苦,但临床缺乏有效的治疗手段。CD147分子是一种广泛表达于人体多种组织的跨膜糖蛋白,属于免疫球蛋白超家族成员。CD147分子在多种肿瘤细胞和组织中高表达,通过诱导基质金属蛋白酶MMPs的分泌促进肿瘤的浸润、转移。同时, CD147与炎症反应如类风湿性关节炎,以及细胞、组织的分化和发育等密切相关。已有研究证明,MMPs通过细胞外基质蛋白的活化和裂解及促炎性细胞因子和趋化因子的作用参与了神经炎症的发生,在神经病理性疼痛的发生机制中起着非常重要的作用。作为MMPs的上游分子,CD147在神经病理性疼痛的发病机制中很可能扮演着重要角色。本研究应用免疫荧光组织化学染色和western blotting方法,观察脊神经选择性结扎(SNL模型)所致神经病理性疼痛条件下,大鼠脊髓(SC)和背根神经节(DRG)中的CD147的表达变化,为进一步研究CD147在神经病理性疼痛发病机制中的作用奠定基础。 1. CD147在神经病理性痛模型大鼠脊髓背角内的表达利用疼痛行为学评估大鼠脊神经选择性结扎模型。雄性SD大鼠80只随机分为2组,假手术组(Sham组)和脊神经选择性结扎组(SNL组)。结果显示:SNL组术后TWL, PWT均明显降低,其中SNL组PWL从术后第1天开始下降,第7天降至最低值,与Sham组相比(P 0.01)有统计学差异;SNL组TWL从术后1天开始缩短,第3日即缩短到最低值,与Sham组相比(P 0.01)有统计学差异。免疫荧光化学染色结果显示:大鼠SNL模型后1、3、7d,术侧和对侧脊髓背角CD147表达均无明显变化,与Sham组相比无明显差异。 2. CD147在神经病理性痛模型大鼠背根神经节内的表达变化免疫荧光化学染色结果显示:大鼠SNL模型后1、3、7d分别观察到术侧背根神经节中CD147表达变化,SNL组从术后第1d即可发现CD147表达开始上调,第3d表达最强。GFAP活化程度同CD147表达相平行,在SNL模型后,背根神经节中GFAP的活化明显增强,在术后3d时活化最为强烈。Sham组各时间点表达无明显差别。 Western blotting结果提示:大鼠SNL模型后1、3、7d,术侧CD147表达明显上调,同免疫荧光组织化学染色结果一致,即术后1d即可出现CD147的表达上调(P 0.05),术后3d表达最强,与对侧和Sham组相比有统计学意义(P 0.01)。结论: 1)大鼠SNL模型后,术侧后肢均产生机械性和热痛敏。机械性痛敏在术后第7d达到高峰,此后略有下降并持续到术后21d。热痛敏在术后第3d达到高峰,此后略有下降并持续到术后21d。 2)正常脊髓背角未见明显CD147表达,SNL模型后连续观察21d,亦未观察到明显表达变化。 3)正常背根神经节内可见少许背景CD147染色,但在SNL模型后,背根神经节中CD147的表达明显上调,且在术后3d时表达最为强烈,术后7天略有下降,但仍维持高水平表达。GFAP活化程度同CD147表达相平行,在SNL模型后,背根神经节中GFAP的活化明显增强,在术后3d时活化最为强烈。 4) CD147在背根神经节中的表达仅局限于细胞间质和神经节被膜,与神经元、星形胶质细胞、内皮细胞及成纤维细胞并无明显重叠。 5)以上结果提示: CD147在背根神经节的表达上调与机械性和热痛敏的产生以及GFAP的活化密切相关,即CD147很可能在神经病理性疼痛的发病机制中起着重要作用。
[Abstract]:Neuropathic pain (Neuropathic pain) refers to the peripheral or central nervous system damage, dysfunction or disease caused by the pain and agony brought with it, but the lack of clinical treatment of.CD147 molecules effectively is a widely expressed in various tissues of human transmembrane glycoprotein belonging to the immunoglobulin superfamily.CD147 molecule high expression in many tumor cells and tissues, through the induction of matrix metalloproteinase MMPs secretion infiltration, promote tumor metastasis. At the same time, CD147 and inflammation such as rheumatoid arthritis, and cells, tissue differentiation and development are closely related. It has been shown that activation and cleavage and proinflammatory cytokine MMPs the extracellular matrix proteins and chemokines involved in nerve inflammation in the pathogenesis of neuropathic pain plays a very important Role. As an upstream molecule of MMPs, CD147 in the pathogenesis of neuropathic pain is likely to play an important role. In this study, immunofluorescence staining and Western blotting method, the observation of selective spinal nerve ligation induced neuropathic pain (SNL model) under the condition of the rat spinal cord and dorsal root ganglion (SC) (DRG) changes in the expression of CD147, which lays a foundation for further study of CD147 in the pathogenesis of neuropathic pain in rats.
1. expression of CD147 in spinal dorsal horn in rats with neuropathic pain model of pain behavior evaluation of selective spinal nerve ligation in rats model. 80 male SD rats were randomly divided into 2 groups, sham operation group (group Sham) and selective spinal nerve ligation group (SNL group). The results showed that: SNL group after operation TWL and PWT were significantly lower in the SNL group, PWL began to decline from the first day after operation, seventh days to a minimum value, compared with the Sham group (P 0.01) had significant difference; SNL group TWL from the beginning 1 days after the operation is shortened, the third day is shortened to a minimum value, compared with the Sham group (P 0.01) there were significant differences.
Immunofluorescence staining showed that there was no significant change in the expression of CD147 in the lateral and contralateral spinal dorsal horn after 1,3,7d in SNL rats, but no significant difference compared with Sham group.
2. CD147 root ganglion in expression of immunofluorescence staining results showed that in the neuropathic pain rat model: 1,3,7d SNL model rats were observed the changes of CD147 expression of dorsal root ganglia, SNL group from 1D after operation can be found in the expression of CD147 began to increase, the expression of 3D.GFAP showed the strongest activation degree between the expression of CD147 in parallel, SNL model, GFAP in dorsal root ganglion activation significantly enhanced after 3D activation was strongest in.Sham group had no significant difference at each time point.
Western blotting showed: 1,3,7d SNL model rats after operation, side CD147 was obviously up-regulated, with immunofluorescence staining results, the postoperative 1D can up-regulated expression of CD147 (P 0.05), the strongest expression after 3D, and on the side and Sham groups have statistical significance (P 0.01).
Conclusion:
1) after the SNL model in rats, both sides of the operative side had mechanical and thermal hyperalgesia. The mechanical pain sensitivity reached the peak at 7d after operation, then decreased slightly and continued to 21d. after the operation. The heat pain reached its peak at 3D after operation, then decreased slightly and continued to 21d. after operation.
2) no obvious CD147 expression was found in the normal dorsal horn of spinal cord, and 21d was observed continuously after SNL model, and no obvious change was observed.
3) normal dorsal root ganglia showed a few background CD147 staining, but in the SNL model, the upregulated expression of CD147 in dorsal root ganglion, and the postoperative 3D expression was the strongest, the 7 day after operation decreased slightly, but still maintain a high level of expression of.GFAP activation with CD147 expression level in. SNL model after dorsal root ganglion in activation of GFAP significantly enhanced after 3D activation was the strongest.
4) the expression of CD147 in dorsal root ganglion is limited to interstitial cells and ganglion capsule. There is no obvious overlap with neurons, astrocytes, endothelial cells and fibroblasts.
5) the above results suggest that the up regulation of CD147 expression in DRG is closely related to the generation of mechanical and thermal pain and the activation of GFAP, that is, CD147 may play an important role in the pathogenesis of neuropathic pain.

【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2010
【分类号】：R363

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