SOCS1在氯胺酮减轻小胶质细胞活化中的机制研究

目的:探讨麻醉药氯胺酮(Ketamine, KET)对暴露于脂多糖(Lipopolysaccharide, LPS)中的小胶质细胞活化水平的影响,并观察细胞因子信号转导抑制因子1(Suppressor of Cytokine Signaling 1, SOCS1)在其中的作用。方法:本研究选用N9小胶质细胞系,将其暴露于浓度为10 ng/mL的LPS中,模拟炎症反应,同时给与浓度为1 m M的KET,采用Western blot、酶联免疫吸附检测(Enzyme-Linked Immunosorbent Assay, ELISA)小RNA干扰和免疫细胞染色等方法,观察KET对暴露于LPS中的小胶质细胞活化水平的影响,及SOCS1分子在其中的作用。结果:将细胞分为3组,分别为正常培养的Control组、LPS组和KET+LPS组,研究发现,将N9小胶质细胞暴露于含10 ng/mL的细胞培养基中24 h后,细胞诱导型一氧化氮合酶(Inducible Nitric Oxide Synthase, i NOS)表达和培养基肿瘤坏死因子α(Tumor Necrosis Factorα, TNF-α)含量显著增加(P0.05),而KET可显著降i NOS蛋白表达和培养基TNF-α含量(P0.05)。随后,将细胞分为5组,分别为Control组、LPS组、KET+LPS组、SOCS1-siRNA+KET+LPS组和乱序siRNA(SC-siRNA)+KET+LPS组,我们发现,LPS可显著增加小胶质细胞TNF-α和白细胞介素1β(Interleukin-1β, IL-1β)的释放、增加SOCS1和核因子κB(Neuclear FactorκB, NF-κB)表达(P0.05),而KET可显著逆转LPS对炎症因子释放和NF-κB表达的影响,并进一步增加SOCS1表达(P0.05),而SOCS1-siRNA显著逆转了KET的上述作用(P0.05),SC-siRNA未对KET产生的上述作用造成显著影响(P0.05)。结论:KET可降低LPS对小胶质细胞的活化作用,上述作用可能通过SOCS1分子介导。     

Stereochemical and structural effects of (2R,6R)-hydroxynorketamine on the mitochondrial metabolome in PC-12 cells

Background

Impairment in mitochondrial biogenesis and function plays a key role in depression and anxiety, both of which being associated with changes in fatty acid and phospholipid metabolism. The antidepressant effects of (R,S)-ketamine have been linked to its conversion into (2S,6S;2R,6R)-hydroxynorketamine (HNK); however, the connection between structure and stereochemistry of ketamine and HNK in the mitochondrial homeostatic response has not yet been fully elucidated at a metabolic level.

Protective effect of erythropoietin against ketamine-induced apoptosis in cultured rat cortical neurons: Involvement of PI3K/Akt and GSK-3 beta pathway

Erythropoietin (EPO) prevents neuronal cell death through the activation of cell survival signals and the inhibition of apoptotic signals in models of neurodegenerative diseases. Here we investigated the neuroprotective effect of EPO in ketamine-induced neurotoxicity in primary cortical neurons. EPO in combination with ketamine greatly increased the cell viability and reduced the number of TUNEL-positive cells. To elucidate a possible mechanism by which EPO exerts its neuroprotective effect, we investigated the phosphoinositide3-kinase pathway using LY294002. The neuroprotection of EPO was prevented by LY294002. Immunoblotting revealed that EPO induced the phosphorylation/activation of Akt and phosphorylation/inactivation of glycogen synthase kinase-3beta (GSK-3β). Moreover, the caspase-3-like activity was increased by addition of ketamine, and decreased by administration of ketamine with EPO. Decreased caspase-3-like activity by administration of ketamine with EPO was restored by LY294002. Our results suggest that PI3K/Akt and GSK-3β pathway are involved in the neuroprotective effect of EPO. You Shang and Yan Wu have contributed equally to this work.     

低体质量小儿心脏手术中应用七氟醚全身麻醉诱导的效果及其对血流动力学的影响

目的:探讨低体质量小儿心脏手术中应用七氟醚全身麻醉诱导的效果及其对血流动力学的影响。方法:选择本院2016年7月-2018年2月期间收治的28例低体质量小儿心脏手术患儿纳入本研究,按照随机数字表法将患儿分为对照组和观察组各14例。对照组采用氯胺酮进行全身麻醉诱导,观察组采用七氟醚吸入麻醉进行全身麻醉诱导。观察两组患儿麻醉诱导敏感性及不良反应等情况,比较两组患儿诱导前、诱导过程中、插管即刻的收缩压(SBP)、舒张压(DBP)、心率(HR)及血氧饱和度(SaO2)水平。结果:观察组患儿哭闹时间、睫毛反射消失时间及疼痛反射消失时间显著低于对照组(P0.05)。观察组患儿不良反应发生率为7.14%(1/14),与对照组的28.57%(4/14)比较差异无统计学意义(P0.05)。诱导过程中及插管即刻,对照组患儿SBP、DBP均低于诱导前和观察组,HR高于诱导前和观察组,SaO_2先降低后升高,且低于观察组(P0.05),观察组患儿SBP低于诱导前,SaO2高于诱导前(P0.05),而DBP、HR与诱导前比较无统计学差异(P0.05)。结论:与氯胺酮相比,采用七氟醚吸入全身诱导麻醉,其对低体质量小儿心脏手术患儿的循环干扰较小,同时在诱导过程中安全稳定,对血流动力学影响较小,容易被患儿所接受。     

四种麻醉药物对食蟹猴麻醉效果分析和选择应用

目的分析比对速眠新、氯胺酮、异氟烷和利多卡因4种不同麻醉药对食蟹猴的麻醉效果。方法总结实际工作中分别使用四种不同麻醉药物对食蟹猴作用的麻醉特点。结果速眠新、氯胺酮、异氟烷均能获得较好的麻醉效果,能满足不同手术、采样需要;局麻药利多卡因对食蟹猴麻醉的实际应用不理想。结论食蟹猴的手术及其他侵犯性操作等都应该考虑生物安全和动物福利要求,实行麻醉,但应根据食蟹猴实验内容要求和不同麻醉药特点选择合适的麻醉方法,确保人员和动物安全,实验结果不受影响。     

Differential effects of chloral hydrate- and ketamine/xylazine-induced anesthesia by the s.c. route

The proper use of anesthetics in animal experimentation has been intensively studied. In this study we compared the use of chloral hydrate (500 mg kg(-1)) and ketamine (167 mg kg(-1)) combined with xylazine (33 mg kg(-1)) by the s.c. route in male Wistar rats. Chloral hydrate and ketamine/xylazine produced a depth of anesthesia and analgesia sufficient for surgical procedures. The decrease of systolic and diastolic blood pressure was of a higher magnitude in rats anesthetized with chloral hydrate than with ketamine/xylazine. The initial microvascular diameter and blood flow velocity did not differ between both agents. On the other hand, ketamine/xylazine reduced the heart rate more intensively than chloral hydrate. Both anesthetics promoted an increase in arterial pCO(2) and a decrease in pH levels compared to unanesthetized animals. The blood glucose levels were of a higher magnitude in rats after ketamine/xylazine anesthesia than after chloral hydrate. In mesenteric arterioles studied in vivo, ketamine/xylazine anesthesia reduced the constrictive effect of noradrenaline and the dilator effect of bradykinin. However, both anesthetics did not modify the vasodilator effect promoted by acetylcholine. Based on our data, we concluded that both anesthetics alter metabolic and hemodynamic parameters, however the use of chloral hydrate in studies of microvascular reactivity in vivo is more appropriate since ketamine/xylazine reduces the responses to vasoactive agents and increases blood glucose levels.     

Responses to innocuous,but not noxious,somatosensory stimulation by neurons in the ferret superior colliculus

The intermediate and deep layers of the superior colliculus (SC) are known for their role in initiating orienting behaviors. To direct these orienting functions, the SC of some animals (e.g., primates, carnivores) is dominated by inputs from the distance senses (vision, audition). In contrast, the rodent SC relies more heavily on non-visual inputs, such as touch and nociception, possibly as an adaptive response to the proximity of dangers encountered during their somatosensory-dominant search behaviors. The ferret (a carnivore) seems to employ strategies of both groups: above ground they use visual/auditory cues, but during subterranean hunting ferrets must rely on non-visual signals to direct orienting. Therefore, the present experiments sought to determine whether the sensory inputs to the ferret SC reveal adaptations common to functioning in both environments. The results showed that the ferret SC is dominated (63%; 181/286) by visual/auditory inputs (like the cat), rather than by somatosensory inputs (as found in rodents). Furthermore, tactile responses were driven primarily from hair-receptors (like cats), not from the vibrissae (as in rodents). Additionally, while a majority of collicular neurons in rodents respond to brief noxious stimulation, no such neurons were encountered in the ferret SC. A small proportion (4%; 13/286) of the ferret SC neurons were responsive to long-duration (> 5s) noxious stimulation, but further tests could not establish these responses as nociceptive. Collectively, these data indicate that the ferret SC is best adapted for the animal's visual/acoustically guided activities and most closely resembles the SC of its phylogenetic relative, the cat.     

右美托咪定与氯胺酮在短小手术麻醉后痛觉过敏及苏醒躁动影响

目的:探究右美托咪定与氯胺酮对短小手术麻醉后痛觉过敏及苏醒躁动的影响。方法:将2017年2月至2019年2月我院手术治疗的93例骨折患儿设为研究对象,将其随机分为A、B、C三组(每组各31例患儿),A组患儿在手术麻醉前30 min静注0.5μg/kg的右美托咪定,B组患儿静注0.5 mg/kg的氯胺酮,C组患儿注射与A、B两组相同剂量的生理盐水,分别使用Ramasy镇静程度量表评估三组患儿苏醒躁动情况,使用Ramasy镇静程度评分评估三组患儿术后镇静情况,使用视觉模拟量表(visual analog scale,VAS)记录三组患儿术后从麻醉药停止使用至VAS评分到达4的时间,而后记录三组患儿使用镇痛药VAS疼痛度降至4以下的实践,最后记录三组患儿术后2 h、4 h及6 h的疼痛度;结果:(1)C组患儿躁动出现率显著高于A组和B组(P0.05),A组和B组对比无差异(P0.05);(2)A组Ramasy镇静程度评分高于B组,B组高于C组(P0.05);(3)A组VAS到达4的时间显著长于B组和C组,且B组也长于C组(P0.05),使用镇痛药后A组VAS评分下降至4的时间显著短于B组和C组,且B组也短于C组(P0.05);(4)患儿术后2 h、4 h、6 h时VAS评分均显著降低,且A组显著低于B组和C组,B组显著低于C组(P0.05)。结论:对实施短小手术患儿应用右美托咪定或氯胺酮能够显著降低患儿术后躁动出现率,同时缓解患儿应用麻醉药后出现的痛觉过敏现象,有助于提高术后安全性。     

氯胺酮复合咪达唑仑全麻在无水乙醇治疗先天性周围血管畸形中的效果评价

目的:观察氯胺酮复合咪达唑仑在无水乙醇治疗儿童先天性周围血管畸形的应用效果。方法:选择择期全麻下行无水乙醇治疗周围血管畸形儿童50例,随机分为两组:氯胺酮复合咪达唑仑组(M组)和丙泊酚持续输注组(P组),M组以咪达唑仑和氯胺酮基础下全麻,P组采用芬太尼镇痛基础下丙泊酚持续静注全麻,观察并记录入室(T_0)、麻醉诱导后5 min(T_1)、手术开始后30、60、90 min(分别为T_2、T_3、T_4)、拔管后5 min(T_5)的MAP与HR,监测脑电双频指数(bispectral index,BIS)并记录BIS40次数、平均注射无水乙醇量、术中输液量、苏醒时间及苏醒后5 min视觉模拟评分(Visual analog scales,VAS)、Ramsay镇静评分结果。结果:(1)与M组相比,P组MAP(T_2～T_54个时间点)、HR(T_1～T_55个时间点)和BIS值(T_3和T_42个时间点)均显著降低,差异有统计学意义(P0.05);三个指标其余时间点两组患儿相比,差异均无统计学意义(P0.05);(2)与M组患儿相比,P组BIS值40的患儿例数以及应用阿托品和麻黄碱例数均显著增加,差异具有统计学差异(P0.05);两组患儿平均无水乙醇注射量比较显著差异无统计学意义(P0.05);(3)与M组相比,P组患儿全麻苏醒拔管后5 min VAS评分显著降低,Ramsay评分显著升高(P0.05);两组患儿不良事件发生率均无统计学差异(P0.05)。结论:氯胺酮复合咪达唑仑下全麻应用于无水乙醇注射治疗儿童周围血管畸形,能维持术中血流动力学平稳,且缩短苏醒时间。