下丘脑弓状核内CBS参与慢性胰腺炎大鼠腹部痛觉过敏的机制研究

发布时间：2018-06-22 15:06

本文选题：下丘脑弓状核 + 慢性胰腺炎　；参考：《苏州大学》2016年硕士论文

【摘要】：目的硫化氢(H2S)是继NO、CO后被发现的第三种气体信号分子,有报道显示初级感觉神经元中的H2S能够参与内脏痛敏的形成,但其在中枢神经系统中的作用却鲜为人知。此研究旨在探讨下丘脑弓状核(ARC)区内H2S内源性合成酶胱硫醚-β-合成酶(CBS)和胱硫醚-γ-裂解酶(CSE)是否参与了慢性胰腺炎腹部痛觉过敏的形成及其可能的机制。方法1.在180-200 g雄性SD大鼠胰管内注射三硝基苯磺酸(TNBS)诱导产生慢性胰腺炎(CP)模型。2.运用von Frey filament(VFF)评测正常大鼠和CP大鼠在ARC微量给药前后的腹部机械痛反应频率。3.运用Western Blotting检测胱硫醚-β-合成酶(CBS)、胱硫醚-γ-裂解酶(CSE)、蛋白激酶Cγ(PKCγ)、总GluN2B和酪氨酸磷酸化的GluN2B(p-GluN2B)在大鼠ARC中的表达水平。结果1.TNBS注射4周后能显著上调ARC内胱硫醚-β-合成酶(CBS)的表达,而没有改变胱硫醚-γ-裂解酶(CSE)的表达。2.与对照组相比,CP虽然并没有改变GluN2B表达的总量,但显著提高了GluN2B的酪氨酸磷酸化水平。CP也显著上调了ARC内PKCγ的表达。3.ARC内微量注射CBS的抑制剂O-(羧甲基)羟胺半盐酸盐(AOAA)能显著降低CP大鼠的腹部疼痛,也翻转了CP大鼠ARC中p-GluN2B和PKCγ上调。4.微量注射PKC特异性抑制剂白屈菜赤碱(CC)能显著减轻CP大鼠的腹部痛觉过敏,并抑制p-GluN2B的增加。5.微量注射NMDA受体拮抗剂D-AP5、GluN2B特异性拮抗剂艾芬地尔均能显著降低CP大鼠的腹部痛觉过敏。6.微量注射H2S供体NaHS能使正常大鼠产生腹部痛觉过敏。结论总之,我们的数据表明,CP引起了ARC内CBS的上调,并可能通过PKCγ介导了GluN2B的活化,参与慢性胰腺炎痛觉过敏。本研究在一定程度上揭示了慢性胰腺炎痛觉过敏产生的中枢机制,并有助于发现慢性胰腺炎疼痛治疗的新靶标。
[Abstract]:Objective hydrogen sulfide (H _ 2S) is the third gas signal molecule found after NO-CO. It has been reported that H _ 2S in primary sensory neurons can participate in the formation of visceral pain sensitivity, but its role in the central nervous system is not well known. The aim of this study was to investigate whether the endogenous synthase cystathithione- 尾 -synthase (CBS) and cystathionine-gamma lyase (CSE) in the hypothalamic arcuate nucleus (ARC) were involved in the development of abdominal hyperalgesia in chronic pancreatitis and its possible mechanism. Method 1. Chronic pancreatitis (CP) model was induced by injecting trinitrobenzene sulfonic acid (TNBS) into the pancreatic duct of 180-200g male SD rats. Von Frey filament (VFF) was used to evaluate the frequency of abdominal mechanical pain response of normal rats and CP rats before and after microadministration of ARC. Western blotting was used to detect the expression levels of cystathithione- 尾 -synthase (CBS), cystathithione-gamma lyase (CSE), protein kinase C 纬 (PKC 纬), total GluN2B and tyrosine phosphorylated GluN2B (p-GluN2B) in rat ARC. Results 1. TNBS significantly upregulated the expression of cystathithione-尾-synthase (CBS) in ARC 4 weeks after injection, but did not change the expression of cystathithione- 纬-lyase (CSE). Compared with the control group, CP did not change the total expression of GluN2B. However, the tyrosine phosphorylation level of GluN2B was significantly increased. CP also significantly up-regulated the expression of PKC 纬 in ARC. 3. Microinjection of CBS inhibitor O- (carboxymethyl) hydroxylamine hemihydrochloride (AOAA) into ARC could significantly reduce abdominal pain in CP rats. The up-regulation of p-GluN2B and PKC 纬 in ARC of CP rats was also reversed. Microinjection of PKC-specific inhibitor, chillerine (CC), significantly alleviated abdominal hyperalgesia and inhibited the increase of p-GluN2B in CP rats. Microinjection of D-AP5GluN2B specific antagonist of NMDA receptor could significantly reduce abdominal hyperalgesia of CP rats. Microinjection of H _ 2S donor NaHS can induce abdominal hyperalgesia in normal rats. Conclusion in conclusion, our data suggest that CP induces the up-regulation of CBS in ARC and may be involved in the hyperalgesia of chronic pancreatitis through PKC 纬 mediated activation of GluN2B. To some extent, this study reveals the central mechanism of hyperalgesia in chronic pancreatitis and helps to identify new targets for the treatment of pain in chronic pancreatitis.
【学位授予单位】：苏州大学
【学位级别】：硕士
【学位授予年份】：2016
【分类号】：R576

【相似文献】