Chronic opioid treatment of neuroblastoma X dorsal root ganglion neuron hybrid F11 cells results in elevated GM1 ganglioside and cyclic adenosine monophosphate levels and onset of naloxone-evoked decreases in membrane K+ currents

Prolongation of the action potential duration of dorsal root ganglion (DRG) neurons by low (nM) concentrations of opioids occurs through activation of excitatory opioid receptors that are positively coupled via G_s regulatory protein to adenylate cyclase. Previous results suggested GM1 ganglioside to have an essential role in regulating this excitatory response, but not the inhibitory (APD-shortening) response to higher (μM) opioid concentrations. Furthermore, it was proposed that synthesis of GM1 is upregulated by prolonged activation of excitatory opioid receptor functions. To explore this possibility we have utilized cultures of hybrid F11 cells to carry out closely correlated electrophysiological and biochemical analyses of the effects of chronic opioid treatment on a homogeneous population of clonal cells which express many functions characteristic of DRG neurons. We show that chronic opioid exposure of F11 cells does, in fact, result in elevated levels of GM1 as well as cyclic adenosine monophosphate (AMP), concomitant with the onset of opioid excitatory supersensitivity as manifested by naloxone-evoked decreases in voltage-dependent membrane K⁺ currents. Such elevation of GM1 would be expected to enhance the efficacy of excitatory opioid receptor activation of the G_s/adenylate cyclase/cyclic AMP system, thereby providing a positive feedback mechanism that may account for the remarkable supersensitivity of chronic opioid-treated neurons to the excitatory effects of opioid agonists as well as antagonists. These in vitro findings may provide novel insights into the mechanisms underlying naloxone-precipitated withdrawal syndromes and opioid-induced hyperalgesia after chronic opiatf addiction in vivo. © 1995 Wiley-Liss, Inc.     

In vivo and in vitro effects of melatonin or ganglioside GT1B on L-cysteine-induced brain mitochondrial DNA damage in mice.

The effects of L-cysteine on mitochondrial DNA (mtDNA) in mouse brain were investigated both in vivoandin vitro. An intracerebroventricular (icv) injection of L-cysteine (1.25 micromol/animal) caused mtDNA damage in brain frontal and central portions of the cortex, broad-spectrum limbic and severe sustained seizures in mice, and increased lipid peroxidation in the whole brain. The L-cysteine-mediated effects were prevented by an intraperitoneal (ip) preinjection of melatonin (20 mg/kg) or an intracerebroventricular preinjection of ganglioside GT1b (90 nmol/animal). Furthermore, in in vitroexperiments, L-cysteine (0.05, 0.5, or 1.0 mM) caused damage to brain mtDNA and increased lipid peroxidation in a concentration-dependent manner when incubated at 37 degrees C for 20 or 60 min with a homogenate prepared from whole mouse brains. However, the mtDNA damage and the increased lipid peroxidation were completely abolished by a cotreatment with melatonin (1.5 mM), a potent scavenger of the hydroxyl radical (*OH), or ganglioside GT1b (60 microM), a potent inhibitor of glutamate-receptor-mediated activation and translocation of protein kinase C and lipid peroxidation. These results suggest that reactive oxygen species including the *OH may be involved in l-cysteine-induced brain mtDNA damage, lipid peroxidation, and development of seizures in mice. Therefore, we concluded that *OH scavengers, such as the pineal hormone melatonin and ganglioside GT1b, can protect against brain mtDNA damage, seizures, and lipid peroxidation induced by reactive oxygen species producers such as L-cysteine.     

神经节苷脂GM_1和GM_3对受体介导的转铁蛋白内吞过程的影响

我们从正常人脑及肝脏中分离得到纯化的神经节苷脂GM_1[Gal-GalNAc-(NeuNAc)Gal-Glc-Cer]和GM_3[NeuNAc-Gal-Glc-Cer],发现外源性GM_1或GM_3的加入能不同程度地抑制SMMC-7721人肝癌细胞的生长,但仅GM_3能抑制转铁蛋白(Tf)促进的细胞生长。进一步的研究发现,GM_3并不影响Tf与细胞膜表面受体的结合,而是抑制了受体介导的Tf内吞过程,内吞囊泡的形成及融合均受到影响。结果提示:GM_3在膜中的嵌入,可能使受体的构象发生变化或影响了膜的流动性,从而抑制Tf的内吞过程。     

GM-1对大鼠视网膜缺血再灌注损伤的保护作用

目的 探讨单唾液酸神经节苷脂（monosialoganglioside,GM-1）对大鼠视网膜缺血冉灌注损伤后视网膜组织及超微结构的保护作用.方法 健康成年Wistar大鼠75只,随机分为3组：正常组5只、NS组35只,GM-1组35只.正常对照组不加任何处理因素,NS组和GM-1组通过升高眼压造成视网膜缺血60 min,分别于缺血前12 h、1 h及缺血结束时3次腹腔注射NS 3mL/kg或GM-1 3mL/kg（10 mg/mL）,并于再灌注0 h（单纯缺血后）、1 h、6 h、12 h、24 h、3 d和7 d共7个时间点取双眼眼球,每时间点5只,HE染色观察视网膜组织结构并测量视网膜内层平均厚度（mean thickhess of the inner retinal layers,MTIRL）,透射电镜观察视网膜超微结构变化.结果 缺血再灌注后,内层视网膜依次表现出水肿、凋亡、萎缩的损伤过程.GM-1可明显减轻其水肿和萎缩的程度,并减少凋亡的发生.结论 GM-1对视网膜缺血再灌注损伤具有明确的保护作用,可能是其有效治疗药物.     

The effect of ganglioside GQ1b on the NMDA receptor signaling pathway in H19-7 cells and rat hippocampus

Gangliosides, sialic acid-containing glycosphingolipids, are related to various synaptic functions in the rat brain. Previously, we investigated the behavioral effects of the ganglioside GQ1b on learning and memory using the Y-maze and Morris water maze test. GQ1b-treated rats showed highly increased memory performance on the Y-maze and the Morris water maze test. In this study, we determined the role of GQ1b on the activation of the N-methyl-d-aspartate (NMDA) receptor signaling pathway in H19-7 rat hippocampal cells and the hippocampus of rats. After 12 h of treatment with GQ1b, the expression levels of NMDA receptor subunit 2A and 2B were increased in H19-7 cells and the hippocampus of rats. In addition, treatment of GQ1b increased the tyrosine phosphorylation of NR2B that may enhance NMDA receptor synaptic activation and enhancement of NMDA receptors. Also, following GQ1b treatment, the phosphorylation of extracellular signal-regulated kinases (ERK1/2) and protein kinase A, a cAMP activated protein kinase (PKA) increased in H19-7 cells and the hippocampus of rats. These increases resulted in an increase in the phosphorylation of cAMP response element binding protein (CREB). These results suggest that GQ1b might facilitate the activation of the NMDA receptor signaling pathway in the hippocampus of rats, an effect which is dependent on ERK1/2, PKA and CREB phosphorylation. Also, these data support our previous result that GQ1b improves the learning and memory of rats.     

神经节苷脂对早产脑白质损伤患儿神经行为发育的影响

目的 探讨神经节苷脂对早产儿脑白质损伤神经行为发育的影响.方法 2005年1月至2009年5月我院NICU收治的早产儿共636例,出生后1周内常规行床边头颅B超检查,确诊为早产儿脑白质损伤的患儿40例,随机分为治疗组与对照组,各20例.治疗组在出生后1周内给予单唾液酸四己糖神经节苷脂钠(GM1)20 mg/d加入葡萄糖液中静脉滴注,1疗程为14 d,根据病情使用1～3个疗程,其他治疗措施同对照组.2组脑白质损伤早产儿均于纠正胎龄40周时进行新生儿行为神经测定,在纠正年龄3、12个月时采用婴幼儿智能发育量表,评估神经系统发育情况.结果 治疗组在纠正胎龄40周时的行为神经测定得分为(38.10±0.91)分,明显高于对照组(36.10±1.59)分(t=4.88,P＜0.05).3个月和12个月时进行的智能发育评估显示智力发育指数(MDI)和心理运动发育指数(PDI),治疗组(3个月MDI:91.66±6.38,PDI:87.11±5.57;12个月MDI:104.10±6.45,PDI:100.46±3.87)均明显高于对照组(3个月MDI:81.07±0.72,PDI:81.90±6.70;12个月MDI:98.45±8.57,PDI:95.91±6.59)(P均＜0.05).结论 GM1对早产儿脑白质损伤神经行为发育有促进作用.

Abstract：

Objective To study the effect of monosialoteterahexosyl ganglioside (GM1) on neurobehavioral development in premature infants with white matter damage. Methods A total of 636premature infants who were hospitalized in NICU of two hospitals from Jan 2005 to May 2009 received routine bedside cranial sonography detection before 1 week-aged. Forty premature infants were diagnosed as being premature white matter damage and divided into the treatment group (20 cases ) and the control group (20 cases) randomly. The cases in the treatment group accepted GM1 20 mg additional to 5% glucose solutionthe iv drip, one time per day,for a cycle of 14 d. 1 -3 cycles were given in accordance with patient's condition. Other treatments were same to the control group. All cases were evaluated by neonatal behavioral and neurological assessment (NBNA) at the rectified age of 40 gestational weeks and by Children's Developmental Center of China (CDCC) test at 3 months-aged and 12 months-aged. Results The NBNA scores of the treatment group (38.10±0.91) were significantly higher than the control group (36.10±1.59) at the rectified age of 40 gestational weeks (P＜0.01). The indexes of mental development(MDI) and psychomotor performance development (PDI) by the CDCC tests in the treating group (3 months-aged MDI:91.66±6.38;PDI:87.11±5.57; 12 months-aged MDI:104.10±6.45; PDI:100.46±3.87) were significantly higher than those in the control group (3 months-aged MDI:81.07±0.72; PDI:81.90±6.70; 12 months-aged MDI:98.45±8.57; PDI:95.91±6.59) at 3 months-aged and 12 months-aged ( P ＜ 0. 05 ). Conclusion GM1 can accelerate the neurobehavioral development in premature infants with white matter damage.     

联合应用神经生长因子和神经节苷脂1对坐骨神经损伤大鼠脊髓神经元的保护作用

目的：了解联合应用神经生长因子(NGF)和神经节苷脂1(GMl)对大鼠周围神经损伤后脊髓神经元的保护作用。方法：选用SD大鼠,分为生理盐水(NS)组、NGF组、GM1组和NGF GM1组,将大鼠坐骨神经造成5mm缺损,术中硅胶管内局部加药、术后大鼠损伤侧小腿肌注药物。术后定期光、电镜观察L4～I6脊髓前角神经元结构变化,测定损伤远段和近段神经传导速度。结果：脊髓运动神经元数目以及神经传导速度4周时,NGF组和GM1组均多于或快于NS组,NGF GM1组则多于或快于NS组、NGF组和GM1组,8周时NGF GM1组、NGF组、GM1组组间无显著性差异但均多于或快于NS组。结论：NGF GM1对周围神经损伤后脊髓运动神经元退变的保护作用与单用NGF或GM1相比,能更早期地发挥作用,并且效果优于单用NGF或GM1。     

神经节苷脂GM1在帕金森氏病症状波动治疗中的应用

目的探讨神经节苷脂GM1治疗帕金森氏病(PD)的疗效和安全性。方法对33例长期服用左旋多巴等治疗帕金森氏病药物后出现症状波动的患者,加用神经节苷脂GM1100mg/d,静脉滴注,每日1次,疗程4周。分别在GM1治疗后2、3、4周对患者进行帕金森病统一评分量表(UPDRS)运动评分及日常生活活动能力(ADL)评分,并观察治疗期间药物的毒副作用。结果33例PD患者在GM1治疗后2、3、4周UPDRS运动评分分别为(23.5±8.9)、(22.8±8.3)和(22.5±9.1),与治疗前(36.7±10.2)比较,均有非常显著性差异(P<0.01);ADL评分分别为(21.4±10.9)、(20.3±9.5)和(20.6±10.2),与治疗前(30.5±12.1)比较,亦均有非常显著性差异(P<0.01)。但治疗2、3、4周的UPDRS运动评分或ADL评分两两比较,无显著性差异(P>0.05)。治疗期间没有观察到明显的毒副作用。结论对长期服用左旋多巴出现症状波动或疗效减退的PD患者,加用GM1治疗,能在一定程度上改善患者的运动功能和日常生活能力。     

Enhancement of liver cell proliferation by GM3 ganglioside

In experiments on rats subjected to partial hepatectomy and experimentally induced hepatitis it is shown that GM₃ ganglioside of equine erythrocytes can enhance liver cell proliferation. The effect was also observed in experiments on a primary hepatocyte culturein vitro; moreover, enhancement of cell proliferation did not depend on the type of sialic acid residues. Translated fromByulleten' Eksperimental'noi Biologii i Meditsiny, Vol. 119, N ^o 4, pp 427–430, April, 1995     

穴位注射对缺氧缺血性脑病患儿恢复期肌张力低下的疗效评价

目的：探讨穴位注射对缺氧缺血性脑病恢复期肌张力低下患儿的治疗效果。方法将96例缺氧缺血性脑病恢复期肌张力低下患儿采用随机数字表法分为治疗组46例,对照组50例,均予以常规综合治疗,治疗组在此基础上加用单唾液酸四己糖神经节苷脂注射液20 m g穴位注射,对照组加用单唾液酸四己糖神经节苷脂注射液20 m g静脉滴注,观察3个月。治疗后采用徒手肌力法评定肌力,表面肌电评估系统评估肱二头肌的恢复情况。结果治疗3个月后两组患儿肱二头肌徒手肌力检查结果显著优于治疗前（P＜0．01）,且治疗组显著优于对照组（P＜0．05）,肌电信号的肌电积分值及均方根值均显著高于治疗前（P＜0．01）,且治疗组显著优于对照组（P＜0．01）。结论在综合康复治疗的基础上辅以局部穴位注射,有助于进一步改善患儿的肌力及肱二头肌的恢复状况。