【摘要】：研究目的：本實驗旨在研究在外周組織炎癥時下丘腦弓狀核（arcuate nucleus ofhypothalamus，ARC）產(chǎn)生中樞敏感化的NMDA受體機制和受體后信號轉(zhuǎn)導途徑。 研究方法：建立完全弗氏佐劑（complete Freund’s adjuvant，CFA）炎癥痛模型；用3管玻璃微電極在體細胞外記錄ARC神經(jīng)元的自發(fā)放電及其對傷害性刺激的反應(yīng)；微電泳MK-801（NMDA受體非競爭性拮抗劑）、PP2（Src家族蛋白酪氨酸激酶抑制劑）、Chelerythrine（蛋白激酶C抑制劑），觀察阻斷NMDA受體或受體后激酶Src和PKC的磷酸化對大鼠ARC神經(jīng)元自發(fā)放電和痛誘發(fā)放電的影響。 研究結(jié)果：（1）CFA炎癥組大鼠ARC神經(jīng)元的自發(fā)放電頻率明顯高于生理鹽水對照組（Saline）大鼠，前者的放電頻率為2.17±0.22Hz，后者為3.63±0.25Hz，兩者比較具有顯著的統(tǒng)計學意義（P0.01）；（2）CFA炎癥組大鼠ARC中痛興奮型神經(jīng)元（PEN, pain-excitatory neuron）的痛誘發(fā)放電頻率明顯高于對照組，前者的放電頻率為3.92±0.19Hz，后者為4.95±0.29Hz，兩者比較具有顯著的統(tǒng)計學意義（P0.01）；（3）微電泳MK-801（3mM，+20~30nA，30s）、PP2（1mM，+20~30nA，10~15min）和Chelerythrine（2mM，，+20~30nA，10~15min）可抑制CFA炎癥組大鼠ARC中PENs的自發(fā)放電，放電頻率分別降低了52.36%（P0.001）、25.5%（P0.05）和25%（P0.05），比較給藥前后放電頻率，皆具有統(tǒng)計學意義；微電泳相同藥物對對照組大鼠的自發(fā)放電頻率不產(chǎn)生影響。（4）微電泳MK-801（3mM，+20~30nA，30s）、PP2（1mM，+20~30nA，10~15min）和Chelerythrine（2mM，+20~30nA，10~15min）可抑制CFA炎癥組大鼠ARC中PENs的痛誘發(fā)放電，放電頻率分別降低了89.15%（P0.001）、77.09%（P0.001）和54.65%（P0.001）；微電泳相同藥物對對照組大鼠ARC中PENs的痛誘發(fā)放電頻率也產(chǎn)生同樣的抑制作用,放電頻率分別降低了90.63%（P0.001）、79.18%（P0.001）和47.06%（P0.01）。 研究結(jié)論：（1）外周CFA炎癥時，ARC作為脊髓上水平痛覺調(diào)節(jié)的一個高位中樞核團，其中的神經(jīng)元可產(chǎn)生敏感化現(xiàn)象，表現(xiàn)為ARC中神經(jīng)元的自發(fā)放電增加，PENs對傷害性刺激的反應(yīng)增強,放電頻率增加。（2）NMDA受體、Src和PKC參與了CFA大鼠ARC中PENs的敏感化，表現(xiàn)為MK-801、PP2和Chelerythrine可以抑制ARC中PENs自發(fā)放電的增加和對傷害性刺激反應(yīng)的增強；（3）NMDA受體、Src和PKC參與了CFA大鼠和對照組大鼠對傷害性刺激的反應(yīng)，表現(xiàn)為MK-801、PP2和Chelerythrine可以抑制其PENs痛誘發(fā)放電的頻率。
[Abstract]:Objective: to investigate the central sensitive NMDA receptor mechanism and postreceptor signal transduction pathway in hypothalamic arcuate nucleus (arcuate nucleus ofhypothalamus,ARC) during peripheral inflammation. Methods: a complete Freund's adjuvant (complete Freund's adjuvant,CFA) model of inflammatory pain was established and the spontaneous discharges of ARC neurons and their responses to noxious stimuli were recorded in vitro with 3-tube glass microelectrodes. Microelectrophoresis MK-801 (NMDA receptor noncompetitive antagonist), PP2 (Src family protein tyrosine kinase inhibitor), Chelerythrine (protein kinase C inhibitor). To observe the effect of blocking phosphorylation of NMDA receptor or post-receptor kinase Src and PKC on spontaneous and pain-induced discharges of ARC neurons in rats. Results: (1) the spontaneous firing frequency of ARC neurons in the CFA inflammatory group was significantly higher than that in the saline control group (2.17 鹵0.22Hz, 3.63 鹵0.25Hz), and the spontaneous firing frequency in the inflammatory group was significantly higher than that in the saline control group (2.17 鹵0.22Hz, 3.63 鹵0.25Hz). There was significant statistical significance between the two groups (P0.01). (2) the frequency of pain-induced discharges of pain-excited neurons (PEN, pain-excitatory neuron) in ARC of rats with CFA inflammation was significantly higher than that of the control group (3.92 鹵0.19 Hz, 4.95 鹵0.29 Hz, respectively). There was significant statistical significance between the two groups (P0.01). (3) Micro-electrophoresis of MK-801 (3mm, 20x30nA, 30s), PP2 (1mm, 20x30nA, 10min) and Chelerythrine (2mm, 20mM, 20k30nA, 10min) inhibited the spontaneous discharge of PENs in ARC of CFA inflammatory rats, and the firing frequency was decreased by 52.36% (P0.001), respectively, and the spontaneous discharge of PENs in ARC was decreased by 52.36% (P0.001) in the inflammatory group of CFA, and the spontaneous discharge frequency was decreased by 52.36% (P0.001). 25.5% (P0.05) and 25% (P0.05). The same drugs had no effect on the spontaneous discharge frequency of the control rats. (4) MK-801 (3mm, 20k30nA, 30s), PP2 (1mm, 20mA, 10min) and Chelerythrine (2mm, 20mA, 30nA), respectively, had no effect on the spontaneous discharge frequency of the control rats. 10~15min could inhibit the pain-induced discharge of PENs in ARC of rats with CFA inflammation, and the firing frequency was decreased by 89.15% (P0.001), 77.09% (P0.001) and 54.65% (P0.001), respectively. The same drugs showed the same inhibitory effect on the pain-induced discharge frequency of PENs in the ARC of the control rats, which decreased by 90.63% (P0.001), 79.18% (P0.001) and 47.06% (P0.01), respectively, and decreased by 90.63% (P0.001), 79.18% (P0.001) and 47.06% (P0.01) respectively. Conclusion: (1) in peripheral CFA inflammation, ARC is a high central nucleus of pain regulation at the level of spinal cord, in which neurons can be sensitized, and the spontaneous discharges of neurons in ARC are increased. The response of PENs to noxious stimuli was enhanced and the discharge frequency was increased. (2) NMDA receptor, Src and PKC were involved in the sensitization of PENs in ARC of CFA rats, which showed MK-801,. PP2 and Chelerythrine could inhibit the increase of spontaneous discharge of PENs and the enhancement of nociceptive stimulation in ARC. (3) NMDA receptors, Src and PKC participated in the responses of CFA rats and control rats to noxious stimuli, which showed that MK-801,PP2 and Chelerythrine could inhibit the frequency of PENs pain-induced discharges.
【學位授予單位】：蘇州大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R338

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