大鼠三叉神经痛慢性缩窄环模型中Na_v1.3、Na_v1.7、Na_v1.8和Na_v1.9的表达及意义

发布时间：2018-03-14 16:50

本文选题：三叉神经痛　切入点：神经病理性疼痛　出处：《山东大学》2016年博士论文　论文类型：学位论文

【摘要】：[研究目的]三叉神经痛(trigeminal neuralgia, TN)是一种神经病理性疼痛。其主要病理变化是三叉神经根的脱髓鞘改变。电压门控钠离子通道(Voltage-gated sodium channels, VGSC)特别是河豚毒素敏感型(tetrodoxin-sensitive, TTX-S)的Nav1.3和Nav1.7及河豚毒素不敏感型(tetrodoxin-resistance, TTX-R)的Nav1.8和Nav1.9在神经病理性疼痛(neuropathic pain, NP)中发挥着重要作用。本研究的目的在于建立稳定的TN大鼠研究模型,探索Nav1.3、Nav1.7、Nav1.8和Nav1.9在神经损伤后的表达变化及其在三叉神经痛中发挥作用的机制。从而寻找治疗三叉神经痛相关的靶点,为临床治疗提供理论依据。[研究方法]研究中我们通过用两根4-0医用铬线结扎眶下神经,结扎线间距约2mm,建立了三叉神经痛的眶下神经(infraorbital nerve, ION)慢性缩窄环术(chronic constriction injury, CCI)模型,并将成年雄性SD大鼠(180～220g)随机分成两组,即假手术组和ION-CCI组。对其机械痛阂进行测定,并采用视频检测的方式,记录大鼠的抓脸次数,以透射电镜观察建模后三叉神经节和眶下神经的组织学改变。此外,通过实时定量聚合酶链反应(Quantitative Polymerase Chain Reaction, QPCR)和免疫荧光双标法检测钠离子通道α亚型基因和蛋白在TG神经元中的表达。[结果]1.行为学检测显示ION-CCI组大鼠机械痛阈在术后4～42天明显下降,在术后12天达到最低,较假手术组有显著性差异(P0.05)。自发行为学视频检测显示ION-CCI组大鼠在术后6～42天抓脸次数明显增多,在术后12天,抓脸次数也达到最高值,较假手术组有显著性差异(P0.05)。2. ION-CCI组大鼠三叉神经节和眶下神经出现不同程度的脱髓鞘改变,而假手术组大鼠则无明显的脱髓鞘现象。3.与假手术组相比,ION-CCI组中Nav1.3的mRNA表达明显上调,Nav1.7、Nav1.8和Nav1.9的mRNA表达则明显下降。4.假手术组和ION-CCI大鼠TG神经元上均有Nav1.3、Nav1.7、Nav1.8和Nav1.9蛋白的表达。在假手术组739/786(94.02%)的TG神经元表达Nav1.3,832/927(89.75%)的TG神经元表达Nay1.7,766/877(87.34%)的TG神经元表达Nay1.8,551/712(77.39%)的TG神经元表达Na,1.9。在ION-CCI组29/801(3.62%)的TG神经元表达Nav1.3,287/869(33.03%)的TG神经元表达Nay1.7,529/874(60.53%)的TG神经元表达Nav1.8,406/742(54.72%)的TG神经元表达Nay1.9。与假手术组相比Navl.3在ION-CCI组大鼠TG神经元上的表达明显增高(P0.01)。Nav1.7、Nav1.8和Nav1.9在ION-CCI组大鼠TG神经元上的表达显著降低(P0.01、P0.01和P0.05)。[结论]我们的研究显示眶下神经慢性缩窄环模型可使大鼠产生类似三叉神经痛的行为学表现和病理学改变。能成为一种较为稳定的离子通道研究模型,用于寻找和研究电压门控钠离子通道亚型在疼痛发生、发展中的功能异常及其分子机制。钠离子通道可能参与了三叉神经痛的调控,寻找出在三叉神经痛中发挥作用的钠离子通道亚型,进而开发特异性的无中枢神经系统及心血管系统副作用的拮抗剂有助于开展三叉神经痛的靶向治疗。
[Abstract]:[Objective] trigeminal neuralgia (trigeminal neuralgia TN) is a kind of neuropathic pain. The main pathological change is the root of trigeminal nerve demyelination. Voltage gated sodium channels (Voltage-gated sodium, channels, VGSC) especially tetrodotoxin sensitive (tetrodoxin-sensitive, TTX-S) is not sensitive to the Nav1.3 and Nav1.7 and tetrodotoxin type (tetrodoxin-resistance, TTX-R) Nav1.8 and Nav1.9 in neuropathic pain (neuropathic pain NP) plays an important role. The purpose of this study is to establish the TN rat model, explore the stability of Nav1.3, Nav1.7, Nav1.8 and Nav1.9 expression in nerve and trigeminal neuralgia mechanism. In order to find the target for the treatment of trigeminal neuralgia related research, provide theoretical basis in research methods.] we through 4-0 with two medical chromium for clinical treatment Infraorbital nerve ligation, ligation line distance of about 2mm, the establishment of the trigeminal neuralgia of the infraorbital nerve (infraorbital nerve, ION) chronic constriction injury (chronic constriction, injury, CCI) model, and adult male SD rats (180 ~ 220g) were randomly divided into two groups: sham operation group and ION-CCI group. The mechanical pain threshold was measured, and the detection of the video recording, rat face catch number, to observe the changes of the trigeminal ganglion and the infraorbital nerve tissue after modeling by transmission electron microscope. In addition, through real-time quantitative polymerase chain reaction (Quantitative Polymerase Chain Reaction, QPCR). The results of]1. behavior detection group ION-CCI showed the mechanical pain threshold of rats on the 4~42 postoperative morning significantly decreased the expression and immunofluorescence method to detect sodium channel alpha gene and protein in TG neurons, after 12 Tianda to the lowest, compared with the sham operation group significantly The difference (P0.05). The spontaneous behavior of video detection showed that ION-CCI rats in 6~42 days after grasping the face number increased significantly, on the 12 day after operation, grasping the face number reached the highest value, there was significant difference compared with the sham operation group (P0.05 group ION-CCI).2. rat trigeminal ganglion and infraorbital nerve appeared different degrees of demyelination, and the rats in the sham operation group had no obvious demyelination.3. compared with sham operation group, Nav1.3 ION-CCI group in mRNA was obviously up-regulated, Nav1.7, Nav1.8 and Nav1.9 mRNA expression decreased TG.4. neurons in sham and ION-CCI rats were Nav1.3, Nav1.7, Nav1.8 expression and the Nav1.9 protein. In the sham operation group (739/786 94.02%) TG (89.75%) expression of Nav1.3832/927 neurons TG neurons expression of Nay1.7766/877 (87.34%) TG (77.39%) expression of Nay1.8551/712 neurons in TG neurons of Na and 1.9. in group ION-CCI 29/8 01 (3.62%) of TG neurons expressed Nav1.3287/869 (33.03%) TG (60.53%) expression of Nay1.7529/874 neurons TG neurons expression of Nav1.8406/742 (54.72%) TG neurons Nay1.9. expression compared with the sham operation group, the expression of Navl.3 in rats of ION-CCI group on the TG neurons were significantly increased (P0.01).Nav1.7, the expression of Nav1.8 and Nav1.9 in ION-CCI group rat TG neurons decreased significantly (P0.01, P0.01 and P0.05). Our research conclusion] infraorbital nerve chronic constriction injury model in rats can cause similar trigeminal neuralgia behaviouristics changes and pathology. To become a more stable ion channel research model, and used to find study of voltage gated sodium channel subtypes in pain, in the development of abnormal function and molecular mechanism of regulation of sodium channel may be involved in the trigeminal neuralgia, find out in pain in the trigeminal nerve function The subtypes of sodium channels used to develop specific antagonists without central nervous system and cardiovascular side effects are helpful for targeted treatment of trigeminal neuralgia.

【学位授予单位】：山东大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R-332;R745.11

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