珠网膜下腔注射2－氯腺苷对脊髓损伤后细胞外钙的影响

目的观察腺苷对大鼠脊髓损伤后脊髓组织细胞外钙的影响,探讨腺苷对脊髓损伤的保护机理。方法脊髓腹侧压迫法造成大鼠T     

Nociceptor activation and protein extravasation induced by inflammatory mediators in human skin.

Protein extravasation (PE) is known to play an important role in inflammatory conditions. In this study we used dermal microdialysis to apply inflammatory mediators (histamine, bradykinin, serotonin) to human skin. Locally induced PE was compared to pain ratings and axon reflex erythema measured simultaneously. Linear microdialysis capillaries (outer diameter 0.4 mm; cut-off 3000 kDa) were inserted intracutaneously at a length of 1.5 cm in the volar forearm of healthy volunteers. The capillaries were perfused with Ringer's solution at a constant flow rate of 4 microl/min. The perfusate was sampled at 15-min intervals and was analysed for total protein concentration. After a baseline of 60 min, the perfusion was switched to inflammatory mediators for 30 min and then back to vehicle again. Sensations evoked by the stimulation were assessed on a visual analogue scale and visible axon reflex erythema was measured planimetrically.Dose-dependent increases in PE could be assessed for all inflammatory mediators tested. Bradykinin (10(-7)M) induced a significant PE, whereas serotonin was effective only at a concentration of 10(-3)M. While serotonin in lower concentrations induced moderate burning pain and an axon reflex flare but no PE, bradykinin provoked PE without pain or axon reflex flare at a concentration of 10(-7)M. Application of histamine similarly evoked PE at lower concentrations as compared to the induction of itch sensation and axon reflex flare.It is concluded that there is no link between nociceptor activation and protein extravasation induced by inflammatory mediators in healthy human skin.     

微透析采样考察玉龙散中乌头碱在大鼠皮肤局部的透皮吸收过程

目的:考察玉龙散中乌头碱的透皮吸收过程。方法:采用HPLC-MS/MS技术建立SD大鼠皮肤微透析液中乌头碱含量的测定方法。通过考察微透析技术中对乌头碱体内回收率的影响因素,确定采样条件。经皮给药后,考察乌头碱的局部透皮吸收过程。结果:本实验成功建立了HPLC-MS/MS测定微透析样品中乌头碱含量的方法。微透析采样流速选择1.5μL·min~(-1),采样间隔为30 min。乌头碱的AUC为18 973.27 h·ng·mL~(-1),MRT为14.97 h,C_(max)为2 976.38 ng·mL~(-1),T_(max)为11.76 h。结论:微透析采样技术可用于玉龙散中乌头碱的透皮吸收过程研究。     

Effect of reserpine on 3-methoxy-4-hydroxyphenylethyleneglycol and 3,4-dihydroxyphenylacetic acid in the hippocampus of depression-model rats: An in vivo microdialysis study

Using in vivo microdialysis, we examined the effect of intraperitoneal injection of reserpine (2 mg/kg) on hippocampal 3-methoxy-4-hydroxyphenylethyleneglycol (MHPG) and 3,4-dihydroxy-phenylacetic acid (DOPAC), two major metabolites of catecholamine. Responses were examined serially for 12 h in the hippocampus of walking-stress-induced depression-model rats, recovery rats and control rats. Control rats showed a rapid rise followed by a gradual fall of free and total MHPG and a delayed increase of DOPAC in response to reserpine. Depression-model rats showed a significantly blunted biphasic response of free and total MHPG as well as blunted monophasic response of DOPAC compared with control rats. Recovery rats also exhibited a blunted fall response of MHPG. Our findings suggest that the vesicle membrane in the central noradrenaline (NA) neurons could be hyposensitive to reserpine in the depression-model rats.     

Escapable and inescapable stress differentially alter extracellular levels of 5-HT in the basolateral amygdala of the rat

The effects of escapable and yoked inescapable electric tailshocks on extracellular levels of serotonin (5-HT) in the basolateral amygdala were measured by in vivo microdialysis. Inescapable, but not escapable, shock increased extracellular 5-HT in the amygdala relative to restrained controls. Basal levels of 5-HT were elevated 24 h after inescapable shock, and previously inescapably shocked subjects exhibited an exaggerated 5-HT response to two brief footshocks. Levels of extracellular 5-HIAA did not follow any particular pattern and were not correlated with the changes in 5-HT.     

Effects of pergolide treatment on in vivo hydroxyl free radical formation during infusion of 6-hydroxydopamine in rat striatum

This study focused on the early neurochemical events involved in 6-hydroxydopamine (6-OHDA) neurotoxicity and the putative neuroprotective effects of pergolide. 6-OHDA in 0.1% ascorbic acid/saline was delivered into rat striatum by means of microdialysis and 2,3-dihydroxybenzoic acid (2,3-DHBA) was measured as an index of hydroxyl free radical formation using salicylate trapping. Infusion of 6-OHDA (2–20 mM) via the dialysis probe for 15 min was associated with an immediate and striking increase in the extracellular levels of 2,3-DHBA and dopamine, and this effect was dose-dependent. An infusion of 10 mM 6-OHDA, equivalent to a direct injection of 4 μg free base, resulted in dopamine overflow with a maximum approx. 200-fold above the baseline. This massive overflow of toxic amounts of dopamine, much greater than expected of reuptake inhibition, seems to be the earliest response of nigrostriatal neurones to 6-OHDA. In rats treated with pergolide mesylate (7 days 0.5 mg/kg/day, i.p.), the average amount of 2,3-DHBA associated with 6-OHDA striatal infusion was significantly smaller than that in controls. This suggests that pergolide treatment leads to an increased ability of striatal tissue to quench hydroxyl radical formation in vivo.     

Chronic electroconvulsive shock and 5-HT autoreceptor activity in rat brain: an in vivo microdialysis study

Summary In vivo microdialysis was used to determine the effects of chronic electroconvulsive shock (ECS), given daily for 10 days, on basal 5-HT levels in rat frontal cortex and hippocampus and on the effect of systemic administration of the 5-HT-la receptor agonist, 8-OH-DPAT (0.2 mg/kg), to reduce 5-HT levels in these areas by activation of somatodendritic autoreceptors. Neither basal 5-HT levels nor the effects of 8-OH-DPAT on 5-HT levels were altered after chronic ECS. The effect of systemic administration of the 5-HT_1A and 5-HT_1B antagonist, (±)-pindolol (10mg/kg), to increase 5-HT levels in hippocampus, was also not affected by chronic ECS.     

Oxytocin Released within the Supraoptic Nucleus of the Rat Brain by Positive Feedback Action is Involved in Parturition-Related Events

Oxytocin is released within the supraoptic nucleus during parturition and suckling. During suckling, such release is important in positive feedback stimulation of oxytocin neurons. We have investigated whether oxytocin released within this hypothalamic nucleus during parturition (1) acts on local receptors to further amplify its own release in a positive feedback manner and (2) is critically involved in the regulation of the delivery process. To examine the effect of the oxytocin antagonist on oxytocin release within the supraoptic nucleus, microdialysates were sampled before and during parturition and either vehicle or the antagonist was infused adjacent to the microdialysis probe directly into the supraoptic nucleus after delivery of the second pup. Intranuclear infusion of an oxytocin receptor antagonist (des-Gly-NH₂d(CH₂)₅[Tyr(Me)²Thr⁴]OVT; 50 ng/0.5 μl) significantly (P<0.01) diminished the parturition-related rise in oxytocin release within the supraoptic nucleus and reduced the number of pups delivered during the first and second 30-min dialysis period compared to vehicle-treated controls. Bilateral infusion of the oxytocin receptor antagonist into the supraoptic nucleus after delivery of the second pup significantly slowed parturition (P<0.05), although the parturition-related rise in plasma oxytocin concentration was unchanged. In addition, the onset of suckling was significantly affected by the antagonist as indicated by fewer live pups and fewer surviving pups with milk in their stomachs 24 hours after parturition (P<0.05). To seek other, periventricular sites of oxytocin action during parturition, oxytocin or the oxytocin antagonist was infused into the lateral cerebral ventricle from the birth of pup 2. Via this route, oxytocin speeded up parturition, but the antagonist was ineffective; thus it appears that periventricular oxytocin-sensitive sites are not normally active in promoting parturition, but can do so. The findings indicate a receptor-mediated positive feedback action of oxytocin on its own release within the supraoptic nucleus during parturition, which seems to be involved in the progress of parturition without significantly affecting circulating oxytocin levels. Oxytocin released within the supraoptic nucleus might be important for the coordinated activation of oxytocin neurons and for the synergistic central and peripheral oxytocin effects involved in the regulation of parturition-related events necessary for the survival of the newborn, including the onset of lactation.     

Effects of isoflurane on in vivo release of acetylcholine in the rat cerebral cortex and striatum

Background : Acetylcholine (ACh) is one of the major excitatory neurotransmitters in the central nervous system, and changes in neural activity induced by anesthesia alter the release of ACh. However, the effects of isoflurane, one of the most widely used volatile anesthetics, on ACh release are not known. The present study attempts to clarify the dose–effect relationship of isoflurane on the in vivo release of ACh in rat brains.
Methods : Changes in the extracellular concentration of ACh and choline in the cerebral cortex and striatum induced by 0.5, 1.0, and 1.5 minimum alveolar concentration (MAC) of isoflurane were determined using a brain microdialysis technique.
Results : In the cortex, the ACh release decreased to 30.8±10.1 (mean±SEM), 10.2±4.1, and 8.1±2.9% of basal value by increasing doses of isoflurane, and in the striatum, to 73.3±4.4, 49.2±4.2, and 40.7±4.5%. The ACh release rapidly recovered control levels with the discontinuance of isoflurane. Choline concentration was not changed significantly by isoflurane except for a decrease to 74.8±4.6% in the striatum by 0.5 MAC. In both the cortex and striatum, the choline concentration decreased with the discontinuance of isoflurane to 70.3±13.3, and 68.2±5.4%, respectively.
Conclusion : The fact that all classic anesthetics reported previously, as well as isoflurane, reduce ACh release supports the hypothesis that the suppression of cholinergic cells is, at least in part, one of the mechanisms of anesthesia.     

An Increase in Extracellular Glutamate is a Sensitive Method of Detecting Ischaemic Neuronal Damage during Cranial Base and Cerebrovascular Surgery. An in Vivo Microdialysis Study

Summary All patients undergoing neurological surgery are at risk for serious complications. Ischaemic damage presenting with hemiparesis or speech difficulties occurs in up to 6% of patients undergoing cerebral bypass procedures and other complicated neurosurgical procedures. Currently available methods for detection of such damage include the use of somatosensory evoked potentials (SSEPs) and electro-encephalography (EEG). Unfortunately, these techniques have false positives and may remain normal in the presence of severe focal neurological deficits. Early detection of potential deficits may prevent or minimize damage through a change in operative or anaesthetic strategy. With the availability of several potential neuroprotective compounds, it is also possible to treat patients at risk of developing ischaemic complications if the individuals are identified early. The excitatory neurotransmitter glutamate is not only a metabolic product, but is also thought to promote ischaemia induced cell injury if released into the extracellular space. It may be a significant parameter for ischaemic brain metabolism. In this report we describe 10 patients who underwent extracranial-intracranial (EC-IC) high flow bypass procedures with routine intra-operative monitoring (IOM) as well as intra-operative in-vivo microdialysis measurement of glutamate. Our aim was to compare intra-operative microdialytic findings and IOM findings with respect to patients' early postoperative clinical courses. Three patients had significant intra-operative glutamate increases indicating ischaemia. Two of these patients awoke with a new neurological deficit (hemiparesis). Routine IOM findings were either normal or showed only transient changes during the time the glutamate levels were high. Our study shows that an increase in extracellular glutamate, as monitored by in-vivo microdialysis, is an excellent early market of neuronal damage. While our glutamate measurements were done off-line, it may be possible to get in future continuous on-line measurements to serve as an early warning system for potential ischaemic damage.