本文選題：丙泊酚 + 藥物遺傳學(xué)��； 參考：《遵義醫(yī)學(xué)院》2017年碩士論文

【摘要】：研究背景與目的:全身麻醉應(yīng)用于臨床已經(jīng)百余年歷史,但全身麻醉藥如何致意識消失及蘇醒的機制仍不清楚。近年來研究發(fā)現(xiàn),藍斑(locus coeruleus,LC)核團內(nèi)去甲腎上腺素能神經(jīng)元具有調(diào)控睡眠和覺醒的重要作用,其遞質(zhì)去甲腎上腺素(Noradrenaline,NE)是中樞覺醒環(huán)路信息傳遞的重要媒介。此外,通過在體、離體電生理等技術(shù)發(fā)現(xiàn)藍斑參與了全身麻醉致意識消失或蘇醒過程。但目前尚未闡明全身麻醉藥如何影響藍斑內(nèi)腎上腺素遞質(zhì)變化以及這種變化與麻醉效應(yīng)的相關(guān)性。本實驗采用大鼠微透析以及藥物遺傳學(xué)的方法,研究靜脈全麻藥丙泊酚麻醉實施過程中,LC-NE遞質(zhì)的釋放對大鼠意識消失和蘇醒調(diào)控的作用與機制。方法:(一)隨機選取健康成年雄性SD大鼠12只,建立LC微透析模型,分別測定對照組和丙泊酚組LC區(qū)域NE含量,每組6只。大鼠神經(jīng)遞質(zhì)收集檢測實驗均在建好模型24h后進行。丙泊酚組先收集2h清醒活動大鼠NE作為基礎(chǔ)水平,尾靜脈給予丙泊酚(11 mg/kg),待大鼠大鼠翻正反射消失(loss of righting reflex,LORR)后,丙泊酚(40 mg/kg/h)持續(xù)泵注1h,待翻正反射恢復(fù)(resumption of righting reflex,RORR)后持續(xù)收集1h,每30min一個樣本;對照組給予等量生理鹽水持續(xù)泵注,利用微透析-高效液相電化學(xué)檢測技術(shù)測定兩組NE含量變化;(二)運用DREADDs技術(shù),通過攜帶特異性激活序列的腺相關(guān)病毒(AAV)轉(zhuǎn)染NE神經(jīng)元,腹腔給予CNO激活神經(jīng)元后觀察LC-NE釋放量的改變以及對丙泊酚麻醉下大鼠行為學(xué)的影響。構(gòu)建特異性激活(h M3Dq)及抑制(h M4Di)LC-NE能神經(jīng)元活性AAV,實驗分組,h M3Dq組(n=6)、h M4Di(n=6)、對照組(n=6)。實驗組在LC區(qū)域微注射AAV 500n L,對照組在LC區(qū)域微注射人工腦脊液(ACSF)500n L,2-3周后,待病毒特異性轉(zhuǎn)染NE能神經(jīng)元,建立LC微透析模型,給予腹腔注射CNO(3mg/kg)激活病毒后,分別測定三組NE的含量變化;(三)行為學(xué)觀察。實驗分兩組,h M3Dq組(n=9)、h M4Di(n=8),實驗組在LC區(qū)域微注射AAV 500n L,對照組在LC區(qū)域微注射人工腦脊液(ACSF)500n L,3周后,待病毒成功轉(zhuǎn)染神經(jīng)元。大鼠尾靜脈置管,丙泊酚(800μg/kg/h)持續(xù)泵注,待LORR時記錄時間為LORR時間,并以同樣劑量維持30min,停止丙泊酚泵注,記錄翻正反射恢復(fù)時間;7d后,大鼠尾靜脈置管1h后,腹腔注射CNO(3mg/kg),30min后,再次記錄LORR、RORR時間。結(jié)果:1.采用微透析技術(shù),對比發(fā)現(xiàn),丙泊酚降低LC細胞外液NE水平(P0.05);丙泊酚降低LC細胞外液DA代謝產(chǎn)物HVA、DOPAC水平,同時降低5-HT水平(P0.05),但對其代謝產(chǎn)物5-HIAA無影響(P0.05);2.DREADDs法特異性激活h M3Dq大鼠,LC細胞外液NE水平升高(P0.05),特異性激活h M4Di大鼠,LC細胞外液NE水平降低(P0.05);3.在丙泊酚麻醉下,LC細胞外液NE水平升高,伴隨RORR時間縮短(P0.05),對LORR時間無影響(P0.05);4.在丙泊酚麻醉下,LC細胞外液NE水平降低,伴隨LORR時間縮短(P0.05),對RORR時間無影響(P0.05)。結(jié)論:1.靜脈給予丙泊酚可以降低腦內(nèi)LC區(qū)域NE遞質(zhì)水平;LC NE水平改變,影響丙泊酚麻醉的誘導(dǎo)及蘇醒時間;2.LC去甲腎上腺素能神經(jīng)元參與全身麻醉致意識消失及恢復(fù)過程。
[Abstract]:Background and objective: general anesthesia has been used in clinical history for more than a hundred years, but the mechanism of consciousness disappearance and revival of general anesthetics is still unclear. In recent years, it has been found that noradrenergic neurons in locus coeruleus (LC) nuclei have an important role in regulating sleep and arousal, and their neurotransmitter norepinephrine (norepinephrine) Noradrenaline, NE) is an important medium for the transmission of information in the central awakening loop. In addition, in vivo and in vitro electrophysiology, it is found that the locus coeruleus participates in the disappearance or revival of consciousness induced by general anesthesia. However, it has not yet been clarified how the general anesthetics affect the changes in epinephrine delivery and the relationship between the changes and the anesthetic effect. In this experiment, microdialysis and pharmacogenetic methods were used to study the effect and mechanism of the release of LC-NE transmitter on the consciousness disappearance and revival of rats during the process of propofol anesthesia. Methods: (1) 12 healthy adult male SD rats were randomly selected and the LC microdialysis model was established, and the control group and the third group were determined respectively. The content of NE in the LC area of poolol group was 6. The rat neurotransmitter collection test was performed after the model 24h was built. The propofol group first collected NE as the base level of 2H conscious rats and propofol (11 mg/kg) to the tail vein (loss of righting reflex, LORR), and propofol (40 mg/kg/h) continuous pump 1H, after resumption of righting reflex, RORR), 1H was collected continuously, one sample per 30min; the control group was given constant pumping of normal saline, and the microdialysis high performance liquid phase electrochemical detection technique was used to determine the change of NE content in two groups; (two) DREADDs technology was used to carry the adeno-related disease with specific activation sequence. Toxic (AAV) transfection of NE neurons, CNO activation neurons in the abdominal cavity to observe the changes in the release of LC-NE and the effect on the behavior of rats under propofol anesthesia. Construction of specific activation (H M3Dq) and inhibition (H M4Di) LC-NE neuron activity AAV, experimental group, H M3Dq group, control group. V 500N L, the control group microinjected the artificial cerebrospinal fluid (ACSF) 500N L in the LC region. After 2-3 weeks, the virus specific transfection of NE neurons, the LC microdialysis model was established. After the intraperitoneal injection of CNO (3mg/kg) activated virus, the changes in the content of the NE were measured in the three groups. (three) the experimental group was divided into two groups. The region was microinjected with AAV 500N L, and the control group microinjected the artificial cerebrospinal fluid (ACSF) 500N L in the LC region. After 3 weeks, the virus was successfully transfected to the neuron. The rat tail vein was inserted into the tube, the propofol (800 mu g/kg/h) was pumped continuously, and the recording time was LORR time to LORR, and the same dose was maintained, and the propofol pump was stopped, and the recovery time of the reflex reflex was recorded; 7 After D, 1H was injected into the caudal vein of rats and CNO (3mg/kg) was injected intraperitoneally. After 30min, LORR and RORR time were recorded again. Results: 1. using microdialysis technique, propofol decreased the NE level of LC extracellular fluid (P0.05), propofol decreased the level of DA metabolites of LC extracellular fluid and reduced the level of LC. No effect (P0.05); 2.DREADDs specific activation of H M3Dq rats, LC cell liquid NE level increased (P0.05), specifically activated h M4Di rats, LC extracellular liquid NE level (P0.05); under propofol anesthesia, the level of extracellular fluid increased, accompanied by time contraction, no effect on the time; 4. under propofol anesthesia, fine The NE level of extracellular fluid decreased and the time of LORR shortened (P0.05), and there was no effect on the time of RORR (P0.05). Conclusion: 1. intravenous propofol can reduce the level of NE transmitters in the LC region of the brain, the change of LC NE level, the induction of propofol anesthesia and the awakening time; 2.LC normethyrofenenal neurons are involved in the consciousness disappearance and recovery of general anesthesia. Cheng.

【學(xué)位授予單位】：遵義醫(yī)學(xué)院
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R614

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