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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
Expression of CD147 in the Spinal Cord and Dorsal Root Ganglia of Rat Neuropathic Pain Model
【作者】 田莉；
【作者基本信息】 第四军医大学 ， 麻醉学， 2010， 硕士
【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】 CD147, a member of the immunoglobulin superfamily, is widely expressed in various tissues, originally identified as a tumor surface protein capable of inducing matrix metalloproteinase (MMPs) expression in fibroblasts. While increased expression of CD147 occurs in many tumors, its expression is not limited to tumor cells. CD147 is a pleiotropic molecule that is critical in fetal development and retinal function and has been shown to play a role in thymic T cell development, as well as many neurological processes ranging from the development of the nervous system to the involvement in spatial learning and plaque formation within Alzheimer?s stricken brains.It was documented that MMPs are invovled in the development and mantainance of neuropathic pain. We proposed a hypothesis that CD147, as a inducer of MMPs, may be directly involved in the development of neuropathic pain, and immune therapy targeted at this molecule might provide effictive means for the treatment of neuropathic pain. To provide evidence that CD147 is invovled in the development of neuropathic pain, immunofluorescence staining and western blotting were used to observe the expression of CD147 in the spinal cord (SC) and dorsal root ganglion (DRG) in a neuropathic pain model --- spinal never ligation model (SNL).1. The expression of CD147 in the SC of the rats after SNL. Behavioral tests were used to assess the success rate of the SNL models. Male SD rats (n=80) were randomly divided into two groups: sham operation group (Sham group) and SNL group. The results showed that the PWT and TWL in SNL group were significantly lower than that in the sham group after the surgery (P < 0.01). The PWT in SNL group reached a peak at day 7 and mantained at least untill day 21. The TWL in SNL group began to decline after 1d of surgery, reached a peak at day 3 and lasted to day 21. Immunofluoresence staining showed that there were no significant expression changes of CD147 between SNL and Sham group after 1,3,7d of SNL in the dorsal horn of the spinal cord.2. The expression of CD147 in the DRG of the rats after SNL. Immunofluorescence staining and western blotting showed that the expression of CD147 was significantly upregulated in the DRG of SNL group, comparing with sham group. The upregulaed expression of CD147 in SNL group reached a peak at day 3 and maintained untill day 21. Activation of GFAP is paralleled with CD147 in the dorsal root ganglion in SNL models. The CD147 stauing was mainly located in the stroma. Double-staining indicated that CD147 staining is not overlaped with neurons, satellite cells, fibroblasts and endothelial cells.Conclusions:The present study indicated that the expression of CD147 was upregulated in the DRGs, but not in the spinal cord, after SNL. The upregulated expression of CD147 is paralleled with the activation of GFAP and the development of mechanical allodynia and thermal hyperalgesia after SNL, suggesting its potential role in the development of neuropathic pain.