重组人红细胞生成素治疗脊髓冲击性损伤的实验研究

[目的]研究重组人红细胞生成素(recombinant human erythropoietin,rh-EPO)对脊髓冲击性损伤的治疗效果。[方法]24只新西兰白兔采用重物坠落的方法造成脊髓冲击性损伤。损伤12 h后,对照组静脉给予生理盐水;小剂量组、中等剂量组和大剂量组分别静脉给予rh-EPO 100、500、1000 IU/kg。损伤后24 h、48 h、1周评估下肢神经功能。损伤后1周时处死动物,脊髓标本进行HE和Caspase-3组化染色,电子显微镜评估超微结构损伤。[结果]EPO治疗组的神经功能评分明显高于对照组,HE染色和电子显微镜显示组织和超微结构损伤明显轻于对照组,Caspase-3阳性细胞数明显少于对照组。中等剂量组和大剂量组的治疗效果无显著性差异。[结论]脊髓冲击性损伤后12 h给予人红细胞生成素可减轻脊髓的组织和超微结构继发性损伤,并可对抗神经细胞凋亡,促进脊髓功能恢复。中等剂量EPO是治疗脊髓损伤的适当选择。     

重组人红细胞生成素与β-七叶皂甙钠治疗大鼠脊髓损伤的实验研究

目的:观察重组人红细胞生成素(rHu-EPO)和β-七叶皂甙钠治疗大鼠脊髓损伤后功能恢复、损伤段神经细胞凋亡情况。方法:30只SD大鼠根据损伤及治疗情况分为4组:空白组、损伤组、治疗A组和治疗B组,观察药物治疗前后大鼠的神经功能行为、红细胞、血红蛋白的变化。损伤后12d处死动物,损伤段脊髓标本做组织学切片,HE染色;做免疫组化检测凋亡因子(包括bcl-2,bax,fas);TUNEL标记凋亡细胞;比较各组间差别。结果:治疗A、B组在用药后,红细胞、血红蛋白数值明显高于治疗前,且与损伤组比较,神经功能有显著提高(P<0.05)。凋亡指标检测:fas、bax、TUNEL显示,损伤组的凋亡阳性细胞明显多于治疗组;bcl-2显示,治疗组的阳性表达明显多于损伤组(P<0.05)。结论:rHu-EPO能抑制脊髓神经细胞凋亡,对损伤脊髓的神经功能具有保护作用,联合应用rHu-EPO和β-七叶皂甙钠效果更明显。     

红细胞生成素在脊髓损伤中抑制细胞凋亡的作用

红细胞生成素(erythropoietin;EPO)是细胞因子超家族的成员之一,除具有促进祖红细胞的增生、分化和成熟的功能外,还具有抗血管痉挛、抗凋亡和抗炎等多种功能。EPO及其受体不仅存在于造血系统中,还存在于中枢和周围神经系统中,EPO除对多种原因引起的脑损伤具有保护作用外,对急性脊髓损伤及其继发性损伤也具有保护作用。近期研究发现其中最主要的保护机制是抑制细胞的凋亡。目前研究认为,抑制神经元细胞凋亡是EPO短期神经保护作用的基础,而EPO的神经营养作用则具有长期保护效果。但其抑制细胞凋亡的细胞内信号传导途径、对神经细胞的营养作用需要进一步的研究。     

重组人促红细胞生成素和甲基强的松龙对急性脊髓损伤大鼠TNF-α表达影响的对比研究

目的 探讨重组人促红细胞生成素(rhEPO)对急性脊髓损伤(SCI)大鼠脊髓组织中促炎因子肿瘤坏死因子(TNF)-α表达的影响,并与甲基强的松龙(MP)进行比较.方法 用改良Allen撞击法制作成年Wistar大鼠SCI模型,随机分为rhEPO组(腹腔注射rhEPO)、MP组(腹腔注射MP)和SCI组(腹腔注射生理盐水对照组).免疫组织化学染色及图像定量分析实验鼠处死后1h、12h、24 h、72 h TNF-α阳性细胞光密度值.结果 rhEPO组、MP组与对照组相比,TNF-α表达明显减弱,差异有统计学意义(P＜0.05);rhEPO组与MP组相比,TNF-α表达无统计学差异(P＞0.05).结论 rhEPO可降低急性SCI大鼠促炎因子TNF-α表达,其效果与MP相当.     

重组人红细胞生成素对大鼠心肌细胞凋亡的影响

心肌梗死后心肌细胞供血不足，产生缺氧损伤，伴随着血管再通，心梗局部再灌，又产生超氧损伤。我们在体外条件下，通过模拟心梗局部的环境变化，探讨重组人红细胞生成素（rHu-EPO）对心肌细胞的缺氧和超氧损伤是否具有保护作用。     

重组人促红细胞生成素对大鼠脊髓损伤后中性粒细胞趋化因子表达的影响及意义

目的 探讨重组人促红细胞生成素(rHuEP())对大鼠脊髓损伤后中性粒细胞趋化因子(CINC-1)表达的影响。方法 SD大鼠102只,随机分为4组,采用改良Allen’s脊髓损伤打击模型,以逆转录一聚合酶链反应(RT-PCR)法测定伤段脊髓组织CINC-1mRNA的表达情况。结果 正常脊髓组织内存在CINC-1mRNA的表达,脊髓损伤后CINC-1mRNA表达迅速增高,伤后6h达到高峰;rHuEPO治疗组脊髓损伤后6、12小时CINC-1mRNA表达明显低于NS治疗组.结论 CINC-1参与继发性脊髓损伤过程,rHuEPO抑制脊髓损伤后CINC-1mRNA的表达,对脊髓继发性损伤可能有保护作用,、     

脊髓损伤后促红细胞生成素对bcl-2的影响

目的 探讨促红细胞生成素（EPO）对大鼠脊髓损伤后伤区脊髓细胞凋亡和神经功能恢复的影响。方法 Wistar大鼠210只，随机分为4组：假手术组、脊髓损伤组、脊髓损伤加重组人EPO治疗组、脊髓损伤加生理盐水治疗组。采用原位末端脱氧核糖核苷酸转移酶介导dUTP标记（TUNEL标记法）检测神经元和少突胶质细胞凋亡，Western blot免疫印迹法和免疫组化染色检测bcl-2表达，免疫组化染色和图像分析方法观察对白质内神经纤维（NF-200染色）的保护作用，通过感觉诱发电位（SSEP）、运动诱发电位（MEP）和大鼠BBB后肢运动功能评分，观察损伤脊髓传导功能的恢复。结果 EPO保护组bcl-2在各时相点的表达明显增高，8h和7d时神经元和少突胶质细胞的TUNEL阳性细胞数明显减少；在7d时白质中NF-200阳性神经纤维明显增多；SSEP和MEP的平均潜伏期和波幅以及BBB功能评分明显提高，与损伤组和生理盐水治疗相比，差异有统计学意义（P〈0．01）。结论 EPO通过上调bcl-2的表达，在抑制脊髓损伤后神经元和少突胶质细胞的凋亡中起到神经保护作用。     

促红细胞生成素治疗急性脊髓损伤

目的 观察促红细胞生成素(EPO)对急性脊髓损伤神经功能恢复的疗效.方法 急性脊髓损伤患者65例,根据用药分为EPO治疗组35例和对照组30例;分别于入院时及治疗后末次随访时对脊髓损伤程度按ASIA2000评分标准进行神经功能评定,观察两组差异;同时监测患者用药前后血常规及血清EPO浓度,记录不良反应.结果 65例患者术后随访1～3年,平均1.7年,末次随访时治疗组患者ASIA运动、触觉、痛觉功能评分为58,2±8.2、78.5±11.5、82.6±13.5,显著优于对照组运动、触觉、痛觉功能评分45.6±6.8、65.5±13.4、68.7±14.7,差异有统计学意义(P<0.05),EPO组治疗过程中未见明显不良反应.结论 促红细胞生成素是治疗急性脊髓损伤安全有效的药物,早期应用EPO对促进患者神经功能的改善与恢复具有积极意义.     

重组人促红细胞生成素对大鼠脊髓损伤后白细胞介素-10表达的影响及意义

目的　探讨重组人促红细胞生成素 (rHuEPO)对大鼠脊髓损伤后白细胞介素 10 (IL 10 )表达的影响。方法　SD大鼠 10 2只 ,随机分为 4组 ,采用改良Allen脊髓损伤打击模型 ,以逆转录 -聚合酶链反应(RT PCR)法测定伤段脊髓组织IL 10mRNA的表达情况。结果　正常脊髓组织内存在IL 10mRNA的表达 ;脊髓损伤后IL 10mRNA表达逐渐增强 ,在伤后 16 8h达高峰 (本实验的最后观察点 ) ;脊髓损伤后 30min注射rHuEPO能明显上调损伤脊髓组织内IL 10mRNA的表达。结论　脊髓损伤后IL 10mRNA表达逐渐增强 ;rHuEPO通过上调IL 10mRNA的表达 ,对脊髓继发性损伤可能有保护作用。     

促红细胞生成素在大鼠脊髓缺血再灌注损伤中的表达及其意义

目的:观察促红细胞生成素(EPO)在大鼠脊髓缺血再灌注损伤(SCII)中的表达和重组人促红细胞生成素(rhuEPO)预处理对再灌注损伤脊髓神经细胞的作用。方法:将W ister大鼠分为正常组、假手术组、rhuEPO处理组和生理盐水对照组;rhuEPO处理组和生理盐水对照组于术前3h腹腔注射rhuEPO和生理盐水,制备大鼠脊髓缺血再灌注损伤模型。以免疫组化和W estern blot法检测脊髓组织中EPO的表达变化;以原位末端脱氧核糖核苷酸转移酶介导dUTP标记法(TUNEL法)检测细胞的凋亡情况。结果:EPO在无损伤脊髓中即有少量的表达,SCII后8h表达显著上调,于12、24h(12h与24h组比较差异无显著性意义,P>0.05)达高峰,伤后3d表达逐渐下调,5d仍保持较高水平。rhuEPO处理组SCII后8h、12h及24h时神经细胞凋亡水平明显低于生理盐水对照组,差异有显著性意义(P<0.01)。结论:在脊髓缺血再灌注损伤中EPO呈现时序性表达变化,可能是机体内源性神经保护的机制之一;EPO预处理能明显抑制SCII后神经细胞的凋亡。     

罗西格列酮对脊髓损伤大鼠神经功能恢复的作用及机制

目的:观察罗西格列酮对脊髓损伤(SCI)大鼠后肢运动功能恢复的作用,探讨其作用机制。方法:75只成年SD大鼠,应用Allen改良法制作大鼠T10SCI模型,随机分为A、B、C三组,每组25只,B、C组于损伤后5min、6h、24h腹腔注射罗西格列酮,C组在腹腔注射罗西格列酮前1h给予G3335,A组于相应时间点腹腔注射等体积生理盐水作为对照组。每组取6只大鼠于伤后1d、7d、2w、4w、6w时对后肢运动功能进行BBB评分;伤后3d每组取4只动物脊髓组织行免疫组织化学染色法检测核转录因子κB(nuclear factor kappa-light-chain-enhancer of activated B cells,NF-κB)的表达;伤后1、3、5、7d和2w每组取3只应用Westernblot法检测脊髓组织中凋亡相关蛋白caspase-3和Bcl-2的表达。结果:伤后1d、7d时3组大鼠BBB评分均为0分,伤后2w开始B组BBB评分高于A组和C组,4w和6w时与A、C组比较有显著性差异(P0.05);伤后3d时三组NF-κB表达均为阳性,但B组平均光密度值明显低于A、C组(P0.05),B组与C组比较无显著性差异(P0.05);伤后各时间点B组caspase-3表达量均低于A组和C组(P0.05),而Bcl-2表达均高于A组和C组(P0.05),其差异均在伤后5d达到高峰,A组与C组同时间点比较无显著性差异(P0.05)。结论:罗西格列酮可促进SCI大鼠神经功能恢复,其机制可能与抑制炎症反应及细胞凋亡有关。     

颈7神经不同平面切断对神经元影响的实验研究

目的 研究颈7神经不同平面切断及不同时期对神经元逆行性退变的影响,为临床选择合适的时间和方法保护受损神经元提供实验依据.方法 36只SD雌性大鼠随机分为三组:对照组、神经根部切断组和神经支部切断组.利用True Blue逆行示踪技术对各组大鼠背根神经节感觉神经元和脊髓前角运动神经元精确计数,并利用神经元特异性烯醇化酶(neuron specific enolase,NSE)免疫荧光双标技术检验标记神经元的活性.结果 术后1周,神经根部切断组和神经支部切断组较对照组无明显的感觉和运动神经元数量变化.术后16周,神经根部切断组和神经支部切断组较对照组感觉和运动神经元减少,其中神经根部切断组较神经支部切断组减少更明显(P＜0.05).且各组True Blue标记神经元全部被NSE免疫荧光双标.结论 颈7神经根部切断和神经支部切断均发生感觉和运动神经元逆行性退变,且不同平面切断对神经元退变的影响有显著性差异.提示臂丛神经根性损伤后早期修复受损神经有利于防止神经元退变.     

Effect of mTOR inhibitor on body weight: from an experimental rat model to human transplant patients

The aim was to study the influence of sirolimus (SRL) on body weight in a rat model and in kidney transplant patients. Wistar rats (15 weeks old) were either treated with vehicle (VEH; n = 8) or SRL (n = 7) 1.0 mg/kg three times per week for 12 weeks. Body mass and food intake were measured weekly. Adipocyte diameter was determined in hematoxylin–eosin stains. The body mass index (BMI) obtained from clinical kidney transplant trials comparing SRL‐based with cyclosporine‐based therapy was analyzed. Animals: SRL produced a decrease of the weight gain curve. At the end of the study, mean body weight in the SRL group was lower than in the VEH group (356 vs. 507 g, P < 0.01) in spite of comparable food intake normalized for body weight was not different. Mean adipocyte diameter was 36 μm in VEH and 25 μm in SRL rats (P = 0.009). Mean SRL blood trough concentration was 38 ng/ml. Kidney transplant patients: Two years after transplantation, BMI was significantly lower in the SRL‐based treatment arm compared to cyclosporine (24.17 ± 2.99 vs. 25.97 ± 5.01 kg/m², P = 0.031). SRL treatment leads to less body mass. Adipocyte cell diameter was reduced in SRL‐treated animals. A possible explanation may be the effects of SRL on metabolic regulation and cell growth.     

Effect of opium dependency on secondary intention wound healing in a rat model: an experimental study

Opium dependency is a social and health problem in some middle eastern countries like Iran. Many of these people may require surgery. This study investigates the effects of opium dependency on histological parameters of secondary intention wound healing in rat. A full‐thickness wound (2 × 2 cm in diameters) was created on the dorsum of two groups of rats, a normal control group and a second group of rat depended to opium (Badawy's method). Several times during 14 days postwounding, the wound was excised with peripheral margins of normal skin and was evaluated for cellular population, reepithelialisation and revascularisation. Results are presented as the mean ± standard error. Data were compared by an unpaired t‐test or analysis of variance. Histological examination of the wound tissue showed evidence of increased population of fibroblasts, decreased recruitment of neutrophile and plateau of macrophage cells in opium depended animals comparing with control group. In the depended animals, reepithelialisation was seen to be enhanced significantly, while prohibiting progression of revascularisation. This study shows that opium dependency enhances reepitheliazation as well as tissue recruitment of fibroblasts; thereby probable enhancement of secondary intention wound healing.     

Effect of naloxone on functional recovery after experimental spinal cord injury in the rat

The effect of the opiate antagonist naloxone on the functional recovery of rats injured with a 10 g-cm impact to the spinal cord at the T-3 level is studied. Sixteen rats were treated with 0.8 mg of naloxone in an intraperitoneal bolus 45 and 120 minutes after injury, 16 rats were given 4 mL of saline instead of naloxone, and 16 rats were neither injured nor treated. To asses weekly the motor recovery of the injured animals, the inclined plane method was employed. After the 10-week assessment period, naloxone-treated animals showed a significantly better performance on the inclined plane than saline-treated animals. Naloxone may be useful for the treatment of spinal cord injury although its mechanism of action remains unknown.     

Activated protein C attenuates intestinal reperfusion-induced acute lung injury: an experimental study in a rat model

BACKGROUND: Activated protein C (APC) is a serine protease with anticoagulant and anti-inflammatory activities. APC has been shown to attenuate local deleterious effects of ischemia/reperfusion (I/R) injury in many organs. We aimed to investigate the effects of APC on lung reperfusion injury induced by superior mesenteric occlusion. METHODS: Male Wistar-Albino rats were allocated into 4 groups: (1) sham-operated group, laparotomy without I/R injury (n = 12); (2) sham + APC group, identical to group 1 except for APC treatment (n = 12); (3) intestinal I/R group, 60 minutes of ischemia followed by 3 hours of reperfusion (n = 12); and (4) I/R + APC-treated group, 100 microg/kg injection of APC intravenously, 15 minutes before reperfusion (n = 12). Evans blue dye was injected into half of the rats in all groups. We assessed the degree of pulmonary tissue injury by measuring activities of oxidative and antioxidative enzymes, as well as nitrate (NO(3)(-))/nitrite (NO(2)(-)) levels, biochemically. We evaluated acute lung injury (ALI) by establishing pulmonary neutrophil sequestration and ALI scoring histopathologically. Pulmonary edema was estimated by using Evans blue dye extravasation and wet/dry ratios. The plasma levels of proinflammatory cytokines and D-dimer were measured. RESULTS: APC treatment significantly reduced activities of oxidative enzymes and nitrate/nitrite levels in the lung tissues, and plasma levels of proinflammatory cytokines and D-dimer, and also significantly increased activities of antioxidative enzymes (P < .05). Pulmonary neutrophil sequestration and ALI scores were decreased significantly with APC administration (P < .05). In addition, APC treatment significantly alleviated pulmonary edema (P < .05). CONCLUSIONS: This study clearly showed that APC treatment significantly attenuated the lung reperfusion injury. Further clinical studies are required to clarify whether APC has a useful role in the reperfusion injury during particular surgeries in which I/R-induced organ injury occurs.     

Effect of different types of textile fabric on spermatogenesis: an experimental study

Summary The effect of different types of textile fabric on spermatogenesis was studied. Twenty-four dogs were divided into two equal groups, one of which wore cotton underpants and the other polyester ones. Seven dogs wearing nothing were used as controls. The underwear was fashioned to fit loosely in the scrotal area so as to avoid its insulating effect. It was worn continuously for 24 months during which the semen character, testicular temperature, hormones (serum testosterone, follicle stimulating hormone, luteinizing hormone, prolactin) and testicular biopsy were examined. The garment was then removed, and the same investigations repeated through another 12 months. The results were analysed statistically. In the polyester group the testicular temperature showed insignificant changes during the period when the pants were worn (P>0.05). By the end of the 24 months there was a significant decrease in sperm count and motile sperms, with an increase in abnormal forms (P<0.001); the testicular biopsy showed degenerative changes. After garment removal the semen character improved gradually to normal in 10 dogs; two remained oligozoospermic. There were insignificant changes (P>0.05) in hormones during the study. In contrast, the cotton and control groups showed insignificant changes (P>0.05) in all the study. The polyester pants thus had a deleterious effect on spermatogenesis in the dogs which was, however, reversible in the majority of cases. The cause of this effect is unknown, but it may be assumed that the electrostatic potentials generated by the polyester fabric play a role in it.     

Analgesic effect of percutaneously absorbed non-steroidal anti-inflammatory drugs: an experimental study in a rat acute inflammation model

Background  

External medication that is absorbed percutaneously may be used to reduce inflammation and relieve pain from acute injuries such as ankle sprains and bruises. The plaster method of percutaneous absorption for non-steroidal anti-inflammatory drugs (NSAIDs) was established in Japan in 1988. However, due to the possibility of a placebo effect, the efficacy of this method remains unclear. This experimental study was conducted to control for the placebo effect and to study the efficacy of the plaster method in relieving pain by using a rat model of inflammation.     

时辰穴位按摩辅治老年卒中后轻度失眠患者的效果

目的探讨时辰穴位按摩对住院老年卒中恢复期心肾不交型轻度失眠患者的疗效,寻求改善此类患者睡眠质量的最佳护理措施实施时间。方法将70例住院老年卒中后轻度失眠患者随机分为观察组(34例)和对照组(36例)。观察组在午时(11:00~13:00)和酉时(17:00~19:00)进穴位按摩,对照组在常规时间,即辰时(7:00~9:00)和未时(13:00~15:00)进行穴位按摩。两组穴位均为少冲穴、复溜穴。用中医睡眠量表(改良型SPIEGEL量表)比较两组患者干预第7、14天的睡眠评分改善情况。结果干预第7天及第14天,观察组患者失眠护理干预治疗效果显著优于对照组(均P0.01)。结论时辰补法穴位按摩有效改善住院老年卒中恢复期心肾不交型轻度失眠患者的睡眠质量。