Radioprotective effect of transferrin targeted citicoline liposomes

The high level of expression of transferrin receptors (Tf-R) on the surface of endothelial cells of the blood–brain-barrier (BBB) had been widely utilized to deliver drugs to the brain. The primary aim of this study was to use transferrin receptor mediated endocytosis as a pathway for the rational development of holo-transferrin coupled liposomes for drug targeting to the brain. Citicoline is a neuroprotective agent used clinically to treat for instance Parkinson disease, stroke, Alzheimer's disease and brain ischemia. Citicoline does not readily cross the BBB because of its strong polar nature. Hence, citicoline was used as a model drug. (Citicoline liposomes have been prepared using dipalmitoylphosphatidylcholine (DPPC) or distearoylphosphatidylcholine (DSPC) by dry lipid film hydration–extrusion method). The effect of the use of liposomes composed of DPPC or DSPC on their citicoline encapsulation efficiency and their stability in vitro were studied. Transferrin was coupled to liposomes by a technique which involves the prevention of scavenging diferric iron atoms of transferrin. The coupling efficiency of transferrin to the liposomes was studied. In vitro evaluation of transferrin-coupled liposomes was performed for their radioprotective effect in radiation treated cell cultures. In this study, OVCAR-3 cells were used as a model cell type over-expressing the Tf-R and human umbilical vein endothelial cells (HUVEC) as BBB endothelial cell model. The average diameter of DPPC and DSPC liposomes were 138 ± 6.3 and 79.0 ± 3.2 nm, respectively. The citicoline encapsulation capacity of DPPC and DSPC liposomes was 81.8 ± 12.8 and 54.9 ± 0.04 μg/μmol of phospholipid, respectively. Liposomes prepared from DSPC showed relatively better stability than DPPC liposomes at 37°C and in the presence of serum. Hence, DSPC liposomes were used for transferrin coupling and an average of 46–55 molecules of transferrin were present per liposome. Free citicoline has shown radioprotective effect at higher doses tested. Interestingly, encapsulation of citicoline in pegylated liposomes significantly improved the radioprotective effect by 4-fold compared to free citicoline in OVCAR-3 but not in HUVEC. Further, citicoline encapsulation in transferrin-coupled liposomes has significantly improved the radioprotective effect by approximately 8-fold in OVCAR-3 and 2-fold in HUVEC cells with respect to the free drug. This is likely due to the entry of citicoline into cells via transferrin receptor mediated endocytosis. In conclusion, our results suggest that low concentrations of citicoline encapsulated in transferrin-coupled liposomes could offer therapeutic benefit in treating stroke compared to free citicoline.     

穴位注射胞二磷胆碱对脑外伤大鼠生长相关蛋白-43 表达的影响

目的观察足三里穴位注射胞二磷胆碱疗法对脑外伤大鼠神经功能及生长相关蛋白(GAP-43)表达的影响。方法成年健康雄性Sprague-Dawley大鼠40 只,除假手术组大鼠8 只(A组)外,其余大鼠采用改良Feeney 法建立大鼠脑损伤模型,造模成功大鼠分为穴位注药组(B 组)、穴位注水组(C 组)、腹腔给药组(D组)和腹腔注水组(E 组)各8 只,造模后每日分别给予足三里穴位注射胞二磷胆碱或生理盐水,腹腔注射胞二磷胆碱或生理盐水,连续14 d。于伤前及伤后第8、14、15、22 天分别行神经功能缺损评分及旷场实验,28 d 后采用免疫组化方法检测大鼠脑组织GAP-43 水平。结果B组大鼠神经缺损评分改善和旷场试验评分均优于C、D、E组(P<0.05)。B组大鼠脑组织GAP-43 阳性表达多于C、D、E组(P<0.05)。结论足三里穴位注射胞二磷胆碱可促进脑外伤大鼠GAP-43 蛋白表达,促进神经功能改善。     

大剂量胞磷胆碱在重型颅脑损伤开颅手术中应用的疗效分析

目的 探讨大剂量胞磷胆碱在重型颅脑损伤开颅手术中防止急性脑肿胀的,临床疗效。方法 36例重型颅脑损伤开颅手术之初,即于切开头皮时,将大剂量胞磷1000mg加入甘露醇250ml内静滴,半小时内滴完。结果 除6例双侧脑挫裂伤严重患者在手术时发生脑肿胀、脑膨出外,其余均未出现脑肿胀、脑膨出现象。结论 重型颅脑损伤开颅手术之初使用大剂量胞磷胆碱,是可以防止急性脑肿胀发生的有效方法之一。     

胞二磷胆碱联合物理疗法治疗弱视的疗效观察

目的探讨胞二磷胆碱联合配镜、遮盖疗法、视近训练治疗弱视的临床效果。方法选择弱视门诊单眼患者8～16岁的268例(268眼),并随机分为观察组和对照组,每组各134例(134眼)。观察组在配镜、遮盖、视近训练物理疗法的基础上联合胞二磷胆碱患眼颞浅动脉旁皮下注射,对照组只用配镜、遮盖疗法物理疗法。随访3个月,观察视力并比较视网膜中央动脉收缩期峰值流速(peak systolic velocity,PSV)及舒张期阻力指数(diastolic resistance index,RI)变化情况。结果观察组134眼总有效率69.40%(93眼),对照组134眼有效率38.81%(53眼)。观察组中轻度、中度、重度弱视患者有效率分别为80.00%、65.52%、43.75%,对照组为50.00%、34.48%、12.50%,2组比较差异均有统计学意义(均为P<0.05)。观察组注射胞二磷胆碱后30min、1h、复查时PSV分别为:(9.61±0.75)mm·s-1、(9.95±0.78)mm·s-1、(9.53±0.65)mm·s-1,较注射前(8.32±0.69)mm·s-1增高(均为P<0.05)。观察组不同时段RI值相比较,差异均无统计学意义(均为P>0.05)。结论胞二磷胆碱联合配镜、遮盖疗法、视近训练治疗大龄弱视儿童能同时提高弱视眼和健眼视力,并且安全有效。     

胞二磷胆碱对神经细胞凋亡保护作用

目的探讨胞二磷胆碱对6-羟基多巴胺所致的神经细胞凋亡的保护作用机制。方法采用流式细胞仪检测神经细胞的凋亡百分比及Bax、Bcl-2蛋白表达。结果将1，0．1，0．01和0．001mmol／L胞二磷胆碱＋6-羟基多巴胺组，与6-羟基多巴胺组比较，凋亡百分比明显降低（P〈0．01）；各组Bax蛋白的表达率明显下降，0．1mmol／L胞二磷胆碱组明显低于对照组；1，0．1和0．01mmol／L胞二磷胆碱组Bcl-2表达率显著升高；Bax／Bcl-2比值下降，0．1mmol／L胞二磷胆碱组Bax／Bcl-2比值低于对照组（P〈0．01）。结论胞二磷胆碱通过减少Bax和增加Bcl-2蛋白的表达。使Bax／Bcl-2比值下降、细胞凋亡减少。发挥其神经保护作用。     

Citicoline/Coenzyme Q10/Vitamin B3 Fixed Combination Exerts Synergistic Protective Effects on Neuronal Cells Exposed to Oxidative Stress

Background: The present study aimed to investigate the rationale and efficacy of using a citicoline, coenzyme Q10 (CAVAQ10) and vitamin B3 fixed combination in combating inflammation and oxidation in neuronal cells exposed to oxidative stress. Methods: HypoE22 cells and isolated hypothalamic specimens were selected as in vitro models to conduct the experiments. The efficacy of citicoline, CAVAQ10, and vitamin B3, with their fixed combination, were assayed after the exposure of hypothalamic cells to hydrogen peroxide (concentration range 1 nM–10 µM), in order to evaluate the biocompatibility of treatments. The activity of neuroprotective and pro-inflammatory factors, namely, brain-derived neurotrophic factor (BDNF), interleukin-6 (IL-6), and tumor necrosis factor-α (TNFα), involved in the neuronal cell damage in neurodegenerative diseases, were assayed in isolated hypothalamus. Results: Neither citicoline, CAVAQ10, nor vitamin B3 significantly altered hypothalamic cell viability, thus suggesting the biocompatibility of single ingredients and fixed combination in the concentration range considered for the study. In the same condition, citicoline and CAVAQ10 were also effective in reducing the gene expression of monoaminoxidase-B, involved in dopamine degradation. However, only citicoline demonstrated an ability to reduce dopamine levels. Conversely, all compounds were effective in reducing the gene expression of IL-6, and TNFα, and in inducing the gene expression of BDNF, with the co-administration of citicoline/CAVAQ10/vitamin B3 being generally more effective than single ingredients. Conclusions: The present findings support the beneficial and synergistic effects of citicoline, CAVAQ10, and vitamin B3 in fixed combination in reducing inflammation and oxidation, and in stimulating neurotrophin production in neuronal cells.     

甘露醇和胞磷胆碱合剂治疗急性脑血管疾病40例

本文对急性脑血管疾病40例(脑梗死37例,脑出血3例;其中男25例,女15例;年龄59±12a)患者用20％甘露醇250mL加胞磷胆碱0.5g静滴治疗,qd,2wk为一个疗程;并与单用甘露醇或右旋糖酐-40组(各为55例、53例)进行比较,经ridit分析,疗效明显优于后2组(P<0.05,及0.01)。作者认为该药可作为治疗脑血管疾病急性期首选药物之一。     

注射用复方胞肌与胞二磷胆碱治疗视神经萎缩的比较

目的：对比观察注射用复方胞肌与胞二磷胆碱治疗视神经萎缩的临床疗效。方法：将６０例视神经萎缩患者随机分为２组,治疗组采用注射用复方胞肌（复方组）,对照组采用胞二磷胆碱（胞液组）,均球后注射给药,２０次为一疗程,将用药前后的视力、视野、视觉诱发电位（ＰＶＥＰ和ＦＶＥＰ）进行比较。结果：临床试验显示复方组各项指标及总疗效均显著优于胞液组（Ｐ〈０．０５）。结论：组成注射用复方胞肌的两药具有较好的协同作用,     

盐酸纳洛酮治疗急性中、重型脑外伤临床研究

目的　探讨盐酸纳洛酮对急性中重型脑外伤患者的治疗作用。方法　将 4 0例急性脑外伤患者随机分为盐酸纳洛酮治疗组 (治疗组 )和胞二磷胆碱治疗组 (对照组 ) ,每组 2 0例。所有患者药物治疗均在脑外伤后 1 2h内 ,在脱水、抗炎、神经营养等常规治疗基础上 ,治疗组加用盐酸纳洛酮 4 .0mg/d ,连续 1 4d为 1疗程。对照组则加用胞二磷胆碱静脉滴注 0 .75g/d。依据意识觉醒时间、GCS评分、GOS评分及TCD等 ,比较两组患者病情恢复情况。结果　急性中、重型脑外伤治疗中 ,治疗组在意识觉醒时间上与对照组相比具有明显统计学意义 (P <0 .0 5 ) ;在不同时期GCS评分上 ,治疗组与对照组相比亦具有明显统计学差异 (P <0 .0 5 ) ;GOS评分治疗组预后明显优于对照组 ,具有非常明显统计学意义 (P <0 .0 1 )。TCD结果显示治疗组缓解脑血管痉挛效果优于对照组。结论　急性脑外伤早期应用盐酸纳洛酮可明显降低颅脑损伤的病残程度 ,提高治疗效果。     

胞磷胆碱治疗脑血栓形成病人57例

本文报道2组脑血栓形成病人各57例,分别用胞磷胆碱和常规药物治疗。总有效率,胞磷胆碱组(84％)略高于常规药物组(77％),无显著性差异(P>0.05)。但治愈率,前者(37％)明显高于后者(14％),差异非常显著(P<0.01)。说明用胞磷胆碱治疗脑血栓形成能提高该病的治愈率,用药愈早疗效愈好。该药有催醒效果,未见明显不良反应。