本文選題：小型豬　切入點：替來他明　出處：《東北農業(yè)大學》2017年碩士論文　論文類型：學位論文

【摘要】：小型豬復合麻醉劑(XFM)是課題組根據小型豬的生理特點和平衡麻醉理論,研制出的一種小型豬專用的復合麻醉劑,具有麻醉誘導時間短、麻醉維持時間適宜、副作用小的優(yōu)點。替來他明和曲馬多作為XFM的成份之一,均作用于動物的中樞神經系統(tǒng),并且都具有確實的鎮(zhèn)痛效果。最新的研究證明:LKB1-AMPK-m TOR信號轉導通路中的關鍵蛋白參與了機體疼痛和神經元興奮性的調節(jié),推測LKB1-AMPK-m TOR信號轉導通路可能參與了麻醉的鎮(zhèn)痛調控。所以本實驗通過給大鼠腹腔注射生理鹽水、鹽酸替來他明和鹽酸曲馬多,利用分子生物學技術檢測藥物注射后不同時間中樞神經系統(tǒng)(大腦皮層、海馬、丘腦、小腦、腦干)中LKB1-AMPK-m TOR信號通路的關鍵蛋白基因的轉錄量和蛋白的磷酸化水平,進而探討他們的鎮(zhèn)痛機制,從而推測XFM的全麻鎮(zhèn)痛機理。將54只SD大鼠隨機分為對照組(n=6)、麻醉組(n=24)和鎮(zhèn)痛組(n=24)。各組大鼠分別腹腔注射生理鹽水、鹽酸替來他明或鹽酸曲馬多后,對照組大鼠在注射生理鹽水后10 min頸椎脫臼處死,麻醉組和鎮(zhèn)痛組根據注射藥物后10 min、20 min、40 min和60 min分為四個亞組(n=6),分別在相應時間點頸椎脫臼處死,并迅速采集大腦皮層、海馬、丘腦、小腦和腦干部位,利用RT-PCR檢測樣品中LKB1 mRNA、AMPKα1 m RNA、AMPKα2 m RNA和4EBP1m RNA的相對轉錄量,利用Western-blot方法檢測各樣品p-LKB1/LKB1、p-AMPK/AMPK和p-4EBP1/4EBP1蛋白磷酸化水平。實驗結果表明:(1)替來他明和曲馬多能使LKB1 m RNA、AMPKα1 m RNA和AMPKα2 m RNA的轉錄量極顯著性升高(p0.01),而抑制4EBP1 m RNA的轉錄量(p0.01)。(2)替來他明和曲馬多能促進LKB1和AMPK的磷酸化水平(p0.01),相反抑制了4EBP1的磷酸化水平(p0.01)。(3)在大鼠的中樞神經系統(tǒng)中,替來他明和曲馬多在海馬、丘腦、小腦以及腦干區(qū)表現出高效的調節(jié)作用,且多為藥物注射后的20 min和40 min。綜上,實驗得出結論:LKB1-AMPK-mTOR信號轉導通路參與了替來他明和曲馬多的麻醉、鎮(zhèn)痛調節(jié),而大鼠的海馬、小腦、丘腦、腦干可能是LKB1-AMPK-m TOR信號通路參與麻醉與鎮(zhèn)痛的靶位;通過激活LKB1-AMPK-mTOR信號通路作用于該藥物相對應的靶蛋白上產生麻醉、鎮(zhèn)痛作用可能是替來他明和曲馬多的作用機制,這一研究結果對于深入揭示小型豬復合麻醉劑的鎮(zhèn)痛和麻醉機理奠定了堅實的基礎。
[Abstract]:The miniature pig compound anesthetic (XFM) is a special compound anesthetic for miniature pigs, which is developed by the research group according to the physiological characteristics and balanced anesthetic theory of miniature pigs. It has the advantages of short induction time and suitable maintenance time. The advantages of small side effects. Teletamine and tramadol, as one of the components of XFM, both act on the central nervous system of animals. Recent studies have shown that the key proteins in the TOR signal transduction pathway of 1: LKB1-AMPK-m participate in the regulation of pain and neuronal excitability. It is speculated that the LKB1-AMPK-m TOR signal transduction pathway may be involved in the analgesic regulation of anaesthesia. Therefore, in this experiment, the rats were injected intraperitoneally with normal saline, teletamine hydrochloride and tramadol hydrochloride. The transcription and phosphorylation of key protein genes of LKB1-AMPK-m TOR signaling pathway in central nervous system (cerebral cortex, hippocampus, thalamus, cerebellum, brainstem) at different time after drug injection were measured by molecular biological techniques. 54 SD rats were randomly divided into control group (n = 6), anesthesia group (n = 24) and analgesia group (n = 24). Rats in each group were given intraperitoneal injection of normal saline, teletamine hydrochloride or tramadol hydrochloride, respectively. The rats in the control group were killed 10 min after injection of normal saline. The anesthetic group and the analgesia group were divided into four subgroups according to the time of 10 minutes, 20 minutes, 40 min and 60 min after injection. The rats were killed at the corresponding time points, and the cerebral cortex was collected rapidly. In hippocampus, thalamus, cerebellum and brain stem, RT-PCR was used to detect the relative transcription of AMPK 偽 1 m RNA-AMPK 偽 2 m RNA and 4EBP1m RNA. The phosphorylation levels of p-LKB1 / AMPK and p-4EBP1/4EBP1 proteins in each sample were detected by Western-blot method. The results showed that teletamine and tramadol could significantly increase the transcription of LKB1 m RNAAMPK 偽 1 m RNA and AMPK 偽 2 m RNA and inhibit the transcription quantity of 4EBP1 m RNA. Lertamine and tramadol could promote the phosphorylation level of LKB1 and AMPK (p0.01), but inhibit the phosphorylation level of 4EBP1 (p0.01) in the central nervous system of rats. Teletamine and tramadol showed high regulatory effects in the hippocampus, thalamus, cerebellum and brain stem area, and most of them were 20 min and 40 mins after drug injection. It is concluded that the LKB1-AMPK-m TOR signal transduction pathway may be involved in the regulation of anesthesia and analgesia by teletamine and tramadol, while the hippocampus, cerebellum, thalamus and brain stem may be the targets of LKB1-AMPK-m TOR signaling pathway. By activating the LKB1-AMPK-mTOR signaling pathway to produce anesthesia on the corresponding target protein of the drug, the analgesic effect may be the mechanism of teletamine and tramadol. The results laid a solid foundation for further revealing the analgesic and anesthetic mechanism of miniature pig compound anesthetic.
【學位授予單位】：東北農業(yè)大學
【學位級別】：碩士
【學位授予年份】：2017
【分類號】：S859.791

【參考文獻】

相關期刊論文 前10條

1 唐雨順;李林;鄒晶;李大鵬;藺東啟;姜勝;范宏剛;;H_2S對噻環(huán)乙胺全麻大鼠血流動力學的影響[J];中國獸醫(yī)雜志;2013年11期

2 唐雨順;李林;鄒晶;李大鵬;藺東啟;姜勝;范宏剛;;噻環(huán)乙胺對大鼠不同腦區(qū)內源性硫化氫濃度的影響[J];中國獸醫(yī)雜志;2013年06期

3 盧德章;范宏剛;于世明;張欒松;馬昆;姜勝;譚麗娟;王洪斌;;替來他明誘導大鼠中樞JUN核蛋白表達的研究[J];中國獸醫(yī)學報;2012年11期

4 葉常紅;楊燕玲;;七氟烷與異氟烷吸入麻醉對肝腎功能影響的比較[J];中國基層醫(yī)藥;2012年10期

5 喻其霞;郭文利;陸雅萍;黃冰;;曲馬多超前鎮(zhèn)痛對乳腺癌根治術后疼痛的影響[J];實用醫(yī)學雜志;2011年11期

6 史鳴;;曲馬多的臨床應用進展[J];中國實用醫(yī)藥;2009年24期

7 張紅勝;周s，

本文編號：1647824