發(fā)布時(shí)間：2018-06-17 22:59

  本文選題：慢性內(nèi)臟痛 + 脊髓背角�。� 參考：《福建醫(yī)科大學(xué)》2015年碩士論文

【摘要】：目的:探究小膠質(zhì)細(xì)胞P2X4受體對(duì)慢性功能性內(nèi)臟痛脊髓中樞敏化作用,為慢性內(nèi)臟痛的藥物治療提供新的作用靶點(diǎn)和治療方向。方法:新生SD大鼠出生第3~21天,每天固定時(shí)間給予3小時(shí)母嬰分離,建立腸易激綜合征(IBS)慢內(nèi)臟痛大鼠模型。通過(guò)檢測(cè)大鼠腹外斜肌對(duì)結(jié)直腸擴(kuò)張的放電反應(yīng)來(lái)評(píng)估IBS模型建立是否成功,并記錄大鼠腹外斜肌在鞘內(nèi)給予P2X4受體阻斷劑5-BDBD、激動(dòng)劑C5-TDS前后對(duì)于結(jié)直腸擴(kuò)張的放電幅值;采用Westernblot方法檢測(cè)大鼠脊髓P2X4受體以及腦神經(jīng)營(yíng)養(yǎng)因子(BDNF)的表達(dá)情況;借助免疫組化的方法檢測(cè)大鼠脊髓背角離子鈣接頭蛋白1(Iba-1)表達(dá)情況來(lái)觀察小膠質(zhì)細(xì)胞的活化狀態(tài);通過(guò)免疫熒光雙標(biāo)方法觀察大鼠脊髓背角P2X4受體與Iba-1蛋白表達(dá)部位是否一致來(lái)確認(rèn)P2X4受體是否在小膠質(zhì)細(xì)胞上表達(dá)。結(jié)果:與正常大鼠相比較,IBS大鼠腹外斜肌對(duì)結(jié)直腸擴(kuò)張刺激反應(yīng)顯著增強(qiáng)(p0.05);鞘內(nèi)給予P2X4受體阻斷劑5-BDBD可抑制IBS和正常大鼠腹外斜肌對(duì)結(jié)直腸擴(kuò)張的放電反應(yīng),同等劑量下,其對(duì)IBS大鼠的阻斷反應(yīng)更明顯(p0.05),而且在IBS大鼠該阻斷劑表現(xiàn)出明顯的劑量依賴效應(yīng);鞘內(nèi)給予P2X4激動(dòng)劑C5-TDS后,IBS和正常大鼠腹外斜肌對(duì)結(jié)直腸擴(kuò)張刺激放電反應(yīng)顯著增強(qiáng),同等劑量下,對(duì)IBS大鼠的激動(dòng)作用更明顯(p0.05),而且在IBS大鼠該激動(dòng)劑表現(xiàn)出明顯的劑量依賴效應(yīng);Western blot結(jié)果顯示,與正常大鼠相比較,IBS大鼠脊髓胸腰段和腰骶段P2X4受體顯著表達(dá)(p0.05)及BDNF蛋白顯著表達(dá)(p0.05);而且鞘內(nèi)給予P2X4受體阻斷劑后,BDNF在IBS大鼠脊髓表達(dá)的抑制程度顯著高于正常大鼠(p0.05);免疫組化結(jié)果顯示,與正常大鼠相比較,IBS大鼠脊髓背角Iba-1蛋白顯著表達(dá)(p0.05),形態(tài)發(fā)生不同程度的肥大,而且鞘內(nèi)給予P2X4受體阻斷劑后,Iba-1在IBS大鼠脊髓背角的表達(dá)抑制程度顯著高于正常大鼠,肥大狀態(tài)被不同程度的減輕;熒光雙標(biāo)結(jié)果顯示,IBS和正常大鼠P2X4受體和Iba-1蛋白在脊髓背角共表達(dá)。結(jié)論:脊髓小膠質(zhì)細(xì)胞P2X4受體對(duì)慢性內(nèi)臟痛中樞具有敏化作用,其機(jī)制可能是通過(guò)激活小膠質(zhì)細(xì)胞釋放腦神經(jīng)營(yíng)養(yǎng)因子導(dǎo)致了慢性內(nèi)臟痛的中樞敏化。
[Abstract]:Aim: to investigate the central sensitization of microglia P2X4 receptor on the spinal cord of chronic visceral pain, and to provide a new target and therapeutic direction for the drug therapy of chronic visceral pain. Methods: Sprague-Dawley (SD) rats were born on the 3rd day of life and were given 3 hours maternal and infant separation at a fixed time every day to establish the model of chronic visceral pain in IBS rats with irritable bowel syndrome (IBS). The success of the establishment of IBS model was evaluated by detecting the discharge response of the ventral oblique muscle to colorectal dilatation, and the discharge amplitude of P2X4 receptor blocker 5-BDBD and the agonist C5-TDS before and after intrathecal administration of P2X4 receptor blocker 5-BDBD were recorded. The expression of P2X4 receptor and brain-neurotrophic factor BDNFin in rat spinal cord were detected by Western blot, and the activation status of microglia was observed by immunohistochemical method. The expression site of P2X4 receptor and Iba-1 protein in spinal dorsal horn of rats was observed by immunofluorescence double labeling method to confirm whether P2X4 receptor was expressed on microglia cells. Results: compared with normal rats, the response of external oblique abdominal muscles to colorectal dilatation was significantly enhanced in IBS rats, and 5-BDBD, a P2X4 receptor blocker, could inhibit the discharge response of IBS and normal rat external oblique muscles to colorectal dilatation, at the same dose, the same dose of P2X4 receptor antagonist 5-BDBD could inhibit the response of IBS and normal rats to colorectal dilatation. The blocking response to IBS was more obvious in IBS rats, and the antagonist showed a dose-dependent effect in IBS rats, and the responses of IBS and normal rat ventral oblique muscles to colorectal dilatation were significantly increased after intrathecal administration of P2X4 agonist C5-TDS. At the same dose, the excitatory effect on IBS rats was more obvious than that on IBS rats, and the agonist showed a significant dose-dependent effect in IBS rats. Compared with normal rats, the expression of P2X4 receptor and BDNF protein in thoracolumbar and lumbosacral segments of IBS rats were significantly higher than those in normal rats, and the inhibition degree of BDNF expression in IBS spinal cord after intrathecal administration of P2X4 receptor blocker was significantly higher than that in normal rats. The results of immunohistochemistry showed that, The expression of Iba-1 protein in the spinal dorsal horn of IBS rats was significantly higher than that of normal rats. The expression of Iba-1 in the spinal dorsal horn of IBS rats was significantly higher than that of normal rats. The expression of Iba-1 in the spinal dorsal horn of IBS rats was significantly higher than that of normal rats, and the expression of Iba-1 in the spinal dorsal horn of IBS rats was significantly inhibited by intrathecal administration of P2X4 receptor blocker. The hypertrophy was alleviated in varying degrees, and the results of fluorescence double labeling showed that P2X4 receptor and Iba-1 protein were co-expressed in the dorsal horn of spinal cord. Conclusion: P2X4 receptor of spinal microglia can sensitize the center of chronic visceral pain by activating microglia to release neurotrophic factor, which may lead to the central sensitization of chronic visceral pain.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R402

【參考文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 林春;IBS樣功能性慢性內(nèi)臟痛的痛覺(jué)敏化作用以及離子型谷氨酸受體的作用[D];福建醫(yī)科大學(xué);2008年

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本文編號(hào)：2032819