Protein kinase C isozymes that mediate enhancement of neurite outgrowth by ethanol and phorbol esters in PC12 cells

Using PC12 cells to study ethanol's effects on growth of neural processes, we found that ethanol enhances NGF- and basic FGF-induced neurite outgrowth. Chronic ethanol exposure selectively up-regulates δ and ε protein kinase C (PKC) and increases PKC-mediated phosphorylation in PC12 cells. Since PKC regulates differentiation, we investigated the role of PKC in enhancement of neurite outgrowth by ethanol. Like ethanol, 0.3–10 nM phorbol 12-myristate, 13-acetate (PMA) increased NGF-induced neurite outgrowth. However, higher concentrations did not, and immunoblot analysis demonstrated that 100 nM PMA markedly depleted cells of β, δ and ε PKC. PMA (100 nM) also down-regulated β, δ and ε PKC in ethanol-treated cells and completely prevented enhancement of neurite outgrowth by ethanol. In contrast, the cAMP analogue 8-bromoadenosine cAMP did not completely mimic the effectsof ethanol on neurite outgrowth, and ethanol was able to enhance neurite formation in mutant PC12 cells deficient in protein kinase A (PKA). These findings implicate β, δ or εPKC, but not PKA, in the neurite-promoting effects of ethanol and PMA. Since chronic ethanol exposure up-regulates δ and ε, but not βPKC, these findings suggest that δ or εPKC regulate neurite outgrowth.     

天麻素对缺血再灌注神经细胞膜的保护作用

用新生Wistar大鼠大脑皮层进行神经细胞培养 ,将培养 8d的神经细胞置于模拟“缺血”状态下 ,5h后神经细胞发生肿胀 ,胞内乳酸脱氢酶 (LDH)漏出增加 ,膜流动性下降、脂质过氧化(LPO)含量增加 ;将经过“缺血”损伤神经细胞置 1 8h模拟“再灌注” ,“再灌注”后细胞数目明显减少 ,LDH的漏出、膜流动性下降和LPO增加继续加重 ,说明细胞损伤进一步加重。经过天麻素孵育后的神经细胞“缺血”或“再灌注”后LDH的漏出及LPO含量明显低于损伤组 ,而膜流动性则明显高于损伤组。表明 :天麻素对体外培养神经细胞的缺血再灌注损伤有保护作用     

NGD及NRF对大鼠坐骨神经缺损修复的电生理学检测

目的 :观察神经生长液 ( NGD)及神经再生素 ( NRF)修复大鼠坐骨神经缺损的效果。方法 :用生理盐水( NS)、NRF桥接以及 NRF桥接 +口服 NGD修复大鼠坐骨神经缺损 ,术后 12周检测大鼠坐骨神经干动作电位传导速度及肌电图。结果 :行为方面 NGD+ NRF组比 NS组恢复早 ,NS组、NRF组、NRF+ NGD组神经干动作电位传导速度的平均值分别为 5 .72 0 m/ s、16 .5 14 m/ s、2 1.310 m/ s。NS组与 NGD+ NRF组的神经干动作电位传导速度经统计学分析有显著性差异 ( P<0 .0 1)。NGD+ NRF组与 NS组的肌电图比较 ,前者的潜伏期短、峰谷值大、运动神经传导速度快。结论 :NGD+ NRF具有良好促进神经再生作用     

实验冻伤大鼠神经纤维的超微结构改变

为探讨冷冻损伤发生、发展及转归的规律，采用健康Ｗｉｓｔａｒ大鼠，分为冻伤组（２０只）和对照组（５只）。电镜下观察大鼠后肢冷冻至－２０℃后即刻、４小时、２４小时及７２小时时周围神经的超微结构改变。冻后即刻，神经髓鞘即出现轻度变性；冻后１～２４小时，有髓神经雪旺细胞及轴突严重变性，而无髓神经变性较轻；冻后７２小时，神经溃变及修复并存，无髓神经较有髓神经易于修复。     

臂丛神经阻滞麻醉进针点选择体会

目的总结锁骨下血管旁间隙臂丛神经阻滞麻醉的体会及效果。方法分析1997年以来行锁骨下血管旁间隙臂丛神经阻滞麻醉100例患者的临床资料。结果无1例出现并发症,麻醉效果佳。结论此法切实可取。     

复方红芪减方对周围神经再生影响的实验研究

目的　研究复方红芪提取液进行减方后对周围神经再生的作用。方法　建立钳夹损伤大鼠双侧坐骨神经的动物模型。按术后每日灌服药物的不同将 40只SD雄性大鼠随机均分为 4组。对照组 :灌服生理盐水 ;复方红芪组 :灌服复方红芪提取液 2ml ;减方组 :灌服复方红芪减方后提取液 2ml;补阳还五汤组 :灌服补阳还五汤 2ml。术后 2周及 4周 ,观察坐骨神经功能指数、运动神经传导速度及单位视野有髓神经纤维计数。结果　坐骨神经功能指数 :红芪组和减方组与对照组、红芪组与减方组的差异均有显著意义 (P >0 .0 5 ) ,且减方组优于红芪组 (P >0 .0 5 )。有髓神经纤维计数 :复方红芪组优于对照组 (P<0 0 1) ,减方组明显优于对照组 (P <0 .0 1) ,而红芪组与减方组间无显著差异。运动神经传导速度 :红芪组、减方组、补阳还五汤组与对照组相比 ,均优于对照组 (P <0 .0 5 ) ,但前 3组间无明显差异。结论　复方红芪减方提取液早期可以促进周围神经再生 ,药效更为专一 ,且优于传统方剂补阳还五汤。     

Brain-derived neurotrophic factor in cerebrospinal fluid of patients with chronic daily headache: relationship with nerve growth factor and glutamate levels

Little has been done to investigate the biochemical basis of chronic daily headache (CDH). Our group has recently demonstrated an increase in the cerebrospinal fluid (CSF) levels of nerve growth factor (NGF) in CDH patients, supporting the involvement of this growth factor in the abnormal processing of head pain in this pathological condition. Other members of the neurotrophin family, especially brain-derived neurotrophic factor (BDNF), have been hypothesized as being involved in the development of chronic head pain in patients affected by CDH, but so far no data are available on this subject. BDNF, NGF and glutamate levels were determined in the CSF of 25 patients affected by CDH with a previous history of migraine. These levels were compared with those of a group of 20 control subjects, for whom the CSF examination and other instrumental investigations excluded diseases of the central and peripheral nervous systems. Significantly higher levels of BDNF, NGF and glutamate were found in CDH patients compared with control subjects (p<0.0001, p<0.0002 and p<0.001, respectively). A significant positive correlation emerged between CSF values of BDNF and those of NGF (r=0.61, p<0.001) and glutamate (r=0.44, p<0.025) in CDH patients. No significant differences were detected in BDNF, NGF and glutamate levels between CDH patients with analgesic overuse and those without. These results support the involvement of BDNF in CDH through the potentiation of glutamatergic transmission involved in the processing of head pain. The significant correlation between BDNF and NGF levels suggests that NGF-mediated up-regulation of BDNF in central sites involved in long-term sensitization plays a key role in persistent head pain in CDH patients. Correspondence to P. Sarchielli     

Microalbuminuria and nerve conduction velocity in type-I diabetics during conventional therapy and during continuous I.V. insulin infusion

Summary The objective of this study was to follow the development of microalbuminuria and nerve conduction velocity under continuous i.v. insulin therapy over a limited period of 4 months. For this purpose, 8 labile type I diabetics were selected (age 33±8 years, duration of diabetes 16±9 years) and treated conventionally with two insulin injections daily over 4 months. Afterwards, the same patients were treated with continuous i.v. insulin infusion and finally again with two injections daily over 4 months each. This procedure allowed each diabetic to serve as his own control. HbA₁, microalbuminuria, nerve conduction velocity and relative refractory period of the ulnar nerve were checked at montly intervals. During the continuous i.v. infusion over 4 months, blood sugar values were significantly lower, glucosuria had disappeared almost completely and the glycosylated hemoglobin had fallen to near normal values. The mean rate of albumin excretion was 16±5 μg/min at rest and 76±26 μg/min during exercise (normal: 3.9±0.4 and 4.8±1.2 μg/min, respectively) and did not change significantly. Nerve conduction velocity in the ulnar nerve rose significantly under i.v. insulin therapy from 47.9±0.6 m/sec to 52±0.6 m/sec. Similarly, the relative refractory period of the same nerve fell significantly from 3.7±0.2 to 1.9±0.1 msec (i.e. to within normal range). It is concluded that functional disturbances of peripheral nerve can regress by improved blood sugar control with continuous i.v. insulin infusion over 4 months. On the other hand, incipient microangiopathy measured as microalbuminuria remains unchanged over the same period of time. If an improvement is at all possible, considerably longer periods of euglycemia are likely to be necessary. Supported by Grant No. 3.964-0.80 from the Swiss National Science Foundation.     

Asystole from tetanic stimulation of the accessory nerve

An asystolic cardiac arrest is reported which occurred at the same time as supramaximal tetanic stimulation over the accessory nerve in order to evoke contractions in the trapezius and sternocleidomastoid muscles. The cause may have been the inadvertent stimulation of one or more of the cranial nerves of the carotid sheath at the base of the skull: the cranial root of the accessory nerve, the vagus, the sino-carotid branch of the glossopharyngeal nerve or the hypoglossal nerve. The most likely culprit, if not the vagus itself, was the cranial root of the accessory nerve which both functionally and anatomically should be seen as an integral part of the vagus. It is suggested that stimulation of any nerve in the carotid sheath should be approached with caution and that a tetanic stimulus to this area might best be avoided.