A Novel Bifunctional Alkylphenol Anesthetic Allows Characterization of γ-Aminobutyric Acid,Type A (GABAA), Receptor Subunit Binding Selectivity in Synaptosomes

Propofol, an intravenous anesthetic, is a positive modulator of the GABA_A receptor, but the mechanistic details, including the relevant binding sites and alternative targets, remain disputed. Here we undertook an in-depth study of alkylphenol-based anesthetic binding to synaptic membranes. We designed, synthesized, and characterized a chemically active alkylphenol anesthetic (2-((prop-2-yn-1-yloxy)methyl)-5-(3-(trifluoromethyl)-3H-diazirin-3-yl)phenol, AziPm-click (1)), for affinity-based protein profiling (ABPP) of propofol-binding proteins in their native state within mouse synaptosomes. The ABPP strategy captured ∼4% of the synaptosomal proteome, including the unbiased capture of five α or β GABA_A receptor subunits. Lack of γ2 subunit capture was not due to low abundance. Consistent with this, independent molecular dynamics simulations with alchemical free energy perturbation calculations predicted selective propofol binding to interfacial sites, with higher affinities for α/β than γ-containing interfaces. The simulations indicated hydrogen bonding is a key component leading to propofol-selective binding within GABA_A receptor subunit interfaces, with stable hydrogen bonds observed between propofol and α/β cavity residues but not γ cavity residues. We confirmed this by introducing a hydrogen bond-null propofol analogue as a protecting ligand for targeted-ABPP and observed a lack of GABA_A receptor subunit protection. This investigation demonstrates striking interfacial GABA_A receptor subunit selectivity in the native milieu, suggesting that asymmetric occupancy of heteropentameric ion channels by alkylphenol-based anesthetics is sufficient to induce modulation of activity.     

Genetic effects on an anesthetic‐sensitive pathway in the brain of drosophila

General anesthetics are known to inhibit the electrically induced escape response of the fruitfly through action within the brain. We examined this response and its sensitivity to anesthetics in several mutants that cause significant disruption of the mushroom body and other structures of the central brain in adult flies. Because we show here that anesthesia sensitivity is influenced by genetic background, we have used a set of congenic mutant lines. Sensitivity to halothane is normal in most of these lines, indicating that the anesthetic target is unaffected by the gross status of the central brain. Thus, for the escape response, anesthetic sensitivity is not a global feature but reflects action at a localized target. Only the mushroom body defect (mud) line showed an increased sensitivity of the escape response to halothane. Sensitivity to two other anesthetics is also perturbed in this line, albeit less dramatically so. The behavior of mud/+ heterozygotes and the comparison of brain anatomy among all the mutant lines imply that the effect of the mud mutation on anesthesia is not via gross alteration of central brain structures. The possibility that an adventitious mutation in the mud line is responsible for the effects on anesthesia is disfavored by the behavior of a heterozygote between two mud alleles. Although we do not yet know whether the mud gene encodes an anesthetic target or influences the functioning of an anesthetic‐sensitive neuron in this pathway, our work indicates that this gene regulates the effects of halothane on a circumscribed pathway. © 2000 John Wiley & Sons, Inc. J Neurobiol 42: 69–78, 2000     

Anesthesia of wild red howler monkeys (Alouatta seniculus) with medetomidine/ketamine and reversal by atipamezole

Wild red howler monkeys (Alouatta seniculus) were translocated during the flooding of the forest at a hydroelectric dam site in French Guiana. For a variety of minor clinical procedures, 96 monkeys were anesthetized with various intramuscular injections of combinations of medetomidine and ketamine. The howler population was composed of healthy animals (42 males and 54 females) of various ages. Medetomidine (150 μg/kg) associated with ketamine (4 mg/kg) gave the best results and was used on 63 animals. The injection rapidly resulted in complete immobilization with good to excellent myorelaxation. The induction stage was quiet, with absence of both corneal and pedal withdrawal reflexes in 57 animals after 2.9 ± 1.4 min. Six animals required an additional injection. Rectal temperature and respiratory and heart rates decreased during anesthesia, whereas relative oxyhemoglobin saturation increased. One death occurred during anesthesia. One abortion and one death also occurred the day following anesthesia but were more probably a result of capture stress. Atipamezole given i.m. at a dose of five times the medetomidine dose 38.4 ± 8.0 min after the anesthetic injection led to standing recovery in 7.1 ± 4.5 min. Spontaneous recovery occurred in 17 animals before the atipamezole injection after an average of 30.6 ± 9.6 min. Total recovery time was shorter in young animals. Medetomidine/ketamine induced good myorelaxation and provided considerably shortened immobilization duration, which are two notable advantages for field studies. We recommend this association for short procedures including minor surgery in red howler monkeys. Am. J. Primatol. 45:399–410, 1998. © 1998 Wiley-Liss, Inc.     

猪全胰十二指肠移植术的麻醉技术及围术期管理探讨

目的-以肠内引流(enteric drainage,ED)和门静脉回流(Portal venous drainage,PVD)术式,建立猪全胰十二指肠移植模型,探讨该术式的麻醉及围术期管理方法,为临床胰腺移植积累经验。方法-供、受体猪各20只,基础麻醉后建立心电监护,硫喷妥钠静脉诱导,暴露声门后,静注司可林气管插管,机控呼吸,术中以芬太尼、咪唑安定、哌库溴铵、硫喷妥钠维持麻醉。直视下右颈总动脉、右颈外静脉穿刺置管,连续监测并记录平均动脉压、心率、经皮脉搏氧饱和度及中心静脉压,测定血糖及胰岛素、胰高血糖素水平。结果-成功建立胰腺移植模型并改进了诱导插管方法,气管插管一次成功率95％。采用静脉复合麻醉,苏醒迅速、安全。无胰期积极扩容可保证新胰灌注后呼吸循环稳定。     

右美托咪定对腰椎全麻手术患者术后疼痛及认知功能的影响

目的:探讨右美托咪定对腰椎全麻手术患者术后疼痛及认知功能的影响。方法:选择2014年3月~2015年12月在我院行腰椎全麻手术的84例患者为研究对象,按照随机数字表法分为对照组(42例)和试验组(42例),患者均常规给予芬太尼及顺式阿曲库铵麻醉诱导,试验组患者在麻醉诱导过程中给予右美托咪定静脉注射,对照组患者仅给予氯化钠注射液静脉注射。分别于术前(T0)、手术开始2 h(T2)、术后24 h(T24)检测血清肾上腺糖皮质激素,采用疼痛视觉模拟评分法(VAS)进行疼痛评定;采用简易智能精神状态量表(MMSE)于术后1d和2d进行认知状态评定,并计算术后认知功能障碍(POCD)发生率;同时观察患者不良反应发生情况。结果:试验组患者T2和T24时肾上腺糖皮质激素水平明显低于T0,T2时试验组患者肾上腺糖皮质激素水平明显低于对照组,差异有统计学意义(P0.05)。试验组患者T2时VAS评分明显低于对照组,差异有统计学意义(P0.05)。术后1d和2d时试验组患者的MMSE评分高于对照组,POCD发生率明显低于对照组;两组患者术后2d时MMSE评分高于术后1d,POCD发生率明显低于术后1d,差异均有统计学意义(P0.05)。两组患者均未见除POCD以外的不良反应。结论:右美托咪定有较强的抗氧化能力,可有效减轻腰椎全麻手术患者的疼痛程度,提高患者的认知功能。     

限制性输液与充分液体复苏在感染性休克患者围手术期麻醉中的应用研究

目的:探究限制性输液与充分液体复苏在感染性休克患者围手术期麻醉中的应用,从而为患者治疗提供相关科学依据。方法:回顾性分析2007年2月至2015年6月期间因感染性休克入院接受治疗的82例患者的临床资料,按输液方式的不同分为研究组(限制性输液)与对照组(充分液体复苏)各41例,观察两组患者转归情况,记录患者心率(HR)、平均动脉压(MAP)、中心静脉压(CVP)、动脉血氧分压(PaO_2)、出血量、总输液量、尿量、术后机械通气时间及ICU住院时间。结果:术后研究组患者出现弥散性血管内凝血(DIC)5例、成人呼吸窘迫综合征(ARDS)3例,2例病情控制不佳者转为多器官功能障碍综合征(MODS),1例死亡,总发生率为26.83%;对照组患者出现DIC 8例,ARDS 7例,MODS 5例,最终出现3例死亡,总发生率为56.10%,两组患者术后转归状况比较,差异有统计学意义(P0.05);研究组患者MAP、CVP、HR、出血量、总输液量及尿量等指标低于对照组,差异有统计学意义(P0.05),研究组患者PaO_2显著高于对照组,差异有统计学意义(P0.05);研究组患者术后机械通气时间及ICU住院时间比对照组短,差异有统计学意义(P0.05)。结论:限制性输液较充分液体复苏能够显著改善感染性休克患者组织血流灌流状况,术中出血量少、术后并发症少,效果更显著,更适合在围手术期麻醉中使用。     

七氟醚联合骶管阻滞麻醉对小儿疝气手术的麻醉效果分析

目的:观察七氟醚联合骶管阻滞麻醉对小儿疝气手术的麻醉效果。方法:选取80例行腹股沟疝气手术患儿,按随机数字表法分为两组,对照组(39例)静脉注射氯胺酮,观察组(41例)先吸入8%七氟醚,然后进行骶管阻滞麻醉,通过观察并记录两组患儿生命体征、麻醉诱导时间、苏醒时间、手术麻醉时间、苏醒期躁动评分(Pediatric anesthesia emergence delirium,PAED)和麻醉诱导期合作量表(Induction Compliance Checklist,ICC)及麻醉期间不良反应情况,评价七氟醚联合骶管阻滞麻醉对小儿疝气手术的麻醉效果。结果:两组切皮后T1、T2时心率(HR)、平均动脉压(MAP)水平均高于T0时的值(P0.05),两组切皮后T1、T2时组间HR、MAP水平相比,无统计学差异(P0.05)。两组切皮后T3时HR、MAP水平基本恢复到T0时的水平。两组切皮前后4个时间点的血氧饱和度(Sp O2)相比,无统计学差异(P0.05)。观察组患儿麻醉诱导时间,苏醒时间均明显短于对照组患儿(P0.05),两组术中麻醉持续时间相比,无统计学差异(P0.05),均能达到期望麻醉时间,观察组患儿PAED评分和ICC评分均低于对照组患儿(P0.05),不良反应组间比较无统计学差异(P0.05)。结论:七氟醚联合骶管阻滞麻醉对小儿疝气手术具有良好的麻醉效果,麻醉诱导快,苏醒快,小儿配合度高,术后躁动少,值得临床推广使用。     

HTEA对全脑缺血/再灌注大鼠脑血流及海马细胞凋亡的影响

目的:研究上胸段硬膜外阻滞(high thoracic epidural anesthesia,HTEA)对大鼠全脑缺血再灌注损伤(global cerebral ischemia,GCI)再灌注期间脑血流及凋亡相关蛋白Bcl-2和Bax的影响。方法:本研究选用成年雄性Wistar大鼠成功进行T4-5间隙硬膜外置管,并建立四血管阻断的全脑缺血模型进行15 min的全脑缺血。根据通过硬膜外导管输注药物不同,随机分为四组:假手术组(Sham,0.9%生理盐水)、假手术-硬膜外组(Sham-HTEA,0.25%布比卡因)、全脑缺血组(GCI,0.9%生理盐水)和硬膜外组(HTEA,0.25%布比卡因)。给药时间从缺血前15 min开始以20μL/h的速度持续输注至再灌注24 h。缺血前至再灌注2 h观察平均动脉压(MAP)和心率(HR),激光多普勒血流仪监测脑血流(cerebral blood flow,CBF),Western-blot检测再灌注24 h海马凋亡蛋白Bcl-2和Bax含量。结果:HTEA组与GCI组相比,缺血期间及再灌注2 h内的MAP和HR无统计学差异,而与Sham组相比,GCI组的MAP缺血时升高,而再灌注时降低(P0.05);再灌注10 min的高灌注期HTEA组CBF明显低于GCI组(123.1%±35.2%vs 177.5%±32.4%,P0.01),再灌注60 min至120 min的低灌注期的大部分时间点HTEA组CBF均高于GCI组(P0.05);再灌注24 h海马组织Bax/Bcl-2比例明显降低(P0.01)。结论:0.25%的布比卡因20μL·h-1连续上胸段硬膜外阻滞可以维持血流动力学稳定,且可改善大鼠全脑缺血再灌注后低灌注期的脑血流量,并减少再灌注24 h海马Bax/Bcl-2比例。     

不同麻醉方法对食管癌手术患者免疫及认知功能的影响

目的:探讨全麻或全麻复合硬膜外麻醉对食管癌手术患者的T细胞水平及术后认知功能的影响。方法:选择2014年1月至2015年12月于我院择期行开胸手术的食管癌患者100例为研究对象,根据手术时间顺序分为观察组(全麻复合硬膜外麻醉)和对照组(全麻),每组50例,观察记录两组患者诱导前、插管时、术中1 h、拔管后的平均动脉压(MAP)、血氧饱和度(Sp O2)和心率(HR);两组患者术前30 min、术后2 h、术后2 d和术后7 d的T细胞亚群水平,包括CD3~+、CD4~+、CD8~+、CD4~+/CD8~+;两组患者术前1 d,术后6 h,术后1 d,术后3 d的认知功能;术后认知功能障碍(POCD)发生率。结果:诱导前观察组和对照组患者的MAP、Sp O2和HR比较,差异均不显著(P0.05),插管时、术中1h和拔管后观察组患者的MAP和HR水平均明显低于对照组(P0.05),而Sp O2明显高于对照组(P0.05)。术后2 h,观察组和对照组的CD3~+、CD4~+、CD8~+、CD4~+/CD8~+值均较术前30 min明显降低(P0.05),但两组间各指标值无显著性差异(P0.05);术后2 d,观察组的CD3~+、CD4~+、CD8~+、CD4~+/CD8~+值均明显高于对照组(P0.05)。术后7 d,两组的T细胞亚群水平均较术前30 min无显著性差异(P0.05)。术后6 h和术后1 d,两组的MMSE评分均较术前1 d明显下降(P0.05),观察组术后1 d、3 d和7 d的MMSE评分均明显高于对照组(P0.05)。术后6 h,观察组的POCD发生率明显低于对照组(P0.05),术后1 d和3 d观察组的POCD发生率低于对照组,但无统计学差异(P0.05)。结论:与单凭全麻比较,全麻复合硬膜外麻醉对食管癌手术患者的T细胞水平及术后认知功能的影响较小,术后恢复快。     

0.5%盐酸罗哌卡因复合芬太尼腰麻在剖宫产术中的临床应用效果

目的:探讨0.5%盐酸罗哌卡因复合芬太尼腰麻在剖宫产术中临床应用效果。方法:选取自2015年01月~2015年05月择期在我院行剖宫产的产妇70例,随机分为观察组和对照组,每组35例。对照组采用0.5%盐酸罗哌卡因进行腰麻,观察组采用0.5%盐酸罗哌卡因复合芬太尼进行腰麻。比较两组的麻醉效果及产妇麻醉指标,记录两组麻醉前后血压及心率变化,并对比两组新生儿出生后1、5、10 min的Apgar评分。结果:观察组麻醉优良率为97.14%,明显高于对照组的85.71%(P0.05)。两组麻醉后30 min血压、心率与麻醉前相比,差异具有统计学意义(P0.05),且两组间比较差异具有统计学意义(P0.05)。观察组痛觉阻滞起效时间、痛觉恢复时间、运动阻滞起效时间、运动恢复时间与对照组相比,差异具有统计学意义(P0.05);观察组新生儿出生后1、5、10min Apgar评分均明显高于对照组(P0.05)。结论:0.5%盐酸罗哌卡因复合芬太尼腰麻应用于剖宫产术,能有效提升麻醉效果,麻醉阻滞起效快,持续时间长,有利于改善新生儿结局,具有临床推广意义。