急性不完全脊髓损伤模型大鼠相关炎症因子的表达

背景：随着现代交通和工矿事业的发展,脊髓损伤成为临床常见的多发病,严重的影响了人类身心健康和生活质量。 目的：检测在大鼠急性脊髓损伤模型中磷酸化信号转导与转录激活因子3（p-STAT3）介导的IL-1β、IL-6、IL-17等炎症因子的表达变化情况,探讨IL-1β、IL-6、IL-17等炎症因子在急性脊髓损伤后炎症反应过程中的作用。 方法：雄性成年SD大鼠75只,随机分为对照组和脊髓损伤1h、6h、24h和72h四个亚组（n=15只）。采用改良Allen氏法建立脊髓损伤模型,对照组仅行椎板全切除术。在建模之后相应的时间点获取损伤段脊髓、脾脏组织,免疫组织化学染色法检测IL-6和IL-17在损伤段脊髓组织中的分布及表达情况,蛋白免疫印迹法检测p-STAT3在损伤段脊髓组织中的表达变化,采用RT-PCR法检测IL-1βmRNA、IL-6mRNA和IL-17AmRNA在脾脏组织中的表达情况。 结果与结论：脊髓损伤后p-STAT3、IL-1β、IL-6和IL-17的表达明显高于对照组（P＜0.05）。损伤后炎症因子表达量立即升高,IL-1β和IL-6在损伤后6h小时到达高峰,随后开始下降;p-STAT3和IL-17在损伤后24h到达高峰,随后开始下降,至损伤后72h表达量仍高于对照组。损伤早期p-STAT3介导的促炎因子IL-1β和IL-6表达增加,可能导致损伤区域炎症级联放大,诱导继发性脊髓损伤;促炎因子IL-17的异常升高,可能在继发性损伤炎症反应中起着重要作用。 中国组织工程研究杂志出版内容重点：肾移植;肝移植;移植;心脏移植;组织移植;皮肤移植;皮瓣移植;血管移植;器官移植;组织工程全文链接：     

白细胞介素17在大鼠哮喘模型肺组织的表达及意义

白细胞介素17(IL-17)是由激活的T淋巴细胞产生的细胞因子，研究证实该细胞因子能增强气道上皮细胞内细胞间粘附分子1(ICAM-1)表达。支气管哮喘不仅气道粘膜存在大量T淋巴细胞浸润，气道内炎性细胞包括中性粒细胞及嗜酸细胞聚集也增多，而ICAM-1在炎性细胞气道聚集中发挥重要作用。     

大鼠脊髓全横断损伤模型的建立

目前用于脊髓全横断的动物模型,常遗有部分未损脊髓组织,局部脊髓损伤范围很大,尤其是破坏了的血运扩大了继发性变性,造成了复杂的局部病理变化,致使难以分析治疗性脊髓伤区内移植的疗效。因此迫切需要一种还原论式的脊髓横断模型,本实验设计了一种切割辅以吸除的方法,可以在局部造成平均约0.6mm的清洁横断区,同时保存脊髓腹、背动脉及脊髓背静脉的大鼠脊髓横断模型,并用H.E.染色法、神经丝(NF)和胶质纤维酸性蛋白(GFAP)的免疫组织化学方法及生物素化的葡萄糖胺(BDA)的追踪方法检验损伤区及其两侧脊髓的组织反应。     

心肌梗死模型大鼠心肌组织中白细胞介素17的表达**☆◆

背景：白细胞介素17在多种心血管疾病中如自身免疫性心肌炎、病毒性心肌炎、扩张型心肌病、不稳定型冠心病等表达增加,在心血管病慢性炎症性过程起到了一定作用。 目的：分析白细胞介素17在心肌梗死大鼠心肌中的表达特点。 方法： 采用液氮冷冻法建立大鼠心肌梗死模型,将造模成功的66只大鼠按照不同的观察时间随机摸球法均分为1,3 ,7,14,28,56 d组,另设对照组8只。采用酶联免疫吸附测定法、免疫荧光激光共聚焦技术及蛋白免疫印迹法检测白细胞介素17的表达。 结果与结论：造模成功率为73%。大鼠心肌梗死的发生发展过程中,白细胞介素17在心肌组织大量表达,且随着时间变化而逐渐升高,在28 d时表达最为显著。说明白细胞介素17作为一种强力的促炎症因子在心肌梗死大鼠心肌组织中大量表达,有可能与心肌梗死后的心室重塑及心律失常的发生有关。 关键词：白细胞介素17;心肌梗死;大鼠;模型;心肌组织 doi:10.3969/j.issn.1673-8225.2012.11.020     

低剂量白细胞介素17A对糖尿病肾病大鼠肾功能保护作用的研究

目的 观察并探讨小剂量白细胞介素17A对糖尿病大鼠肾脏的保护作用.方法 雄性SD大鼠45只,随机分3组:正常对照组、DN组及IL-17A组.DN组及IL-17A组均以高脂高糖饮食联合腹腔注射链脲佐菌素建立糖尿病肾病大鼠模型.IL-17A组给予小剂量IL-17A重组蛋白,连续给药12周后,下腔静脉取血,检测糖化血红蛋白、肌酐、尿素氮等;将大鼠放入水平代谢笼中收集各组大鼠24h尿液,测定大鼠24h尿蛋白;实时荧光定量PCR检测肾组织炎症因子表达;Western blot测定IL-17A蛋白表达情况.结果 DN组及IL-17A组尿蛋白、HbA1c、肌酐、尿素氮均显著高于正常对照组(P＜0.05);IL-17A组尿蛋白、肌酐、尿素氮均显著低于DN组(P＜0.05).DN组大鼠在注射IL-17A后促炎因子表达显著减少,抑制性炎症因子表达显著降低.糖尿病肾病模型大鼠IL-17A蛋白显著减少.结论 IL-17A具有改善糖尿病肾病大鼠肾功能作用,并有可能逆转糖尿病肾病,是治疗糖尿病肾病的有效靶点.     

脊髓全横断大鼠模型的构建

背景：脊髓全横断模型在造模时常难以保证神经纤维的完全离断。 目的：构建大鼠脊髓全横断损伤模型。 方法：将大鼠随机分为模型组和假手术组。模型组构建脊髓T10节段全横断模型;假手术组动物仅打开椎管与硬脊膜而后缝合,但不损伤脊髓。建模后1,3,5,7 d分别进行BBB评分以评估后肢运动功能,检测其体感诱发电位和运动诱发电位来评估神经传导通路的完整性,并行形态学观察来评估脊髓肉眼观病理形态。 结果与结论：与假手术组相比,模型组大鼠在建模后1,3,5,7 d时,其BBB评分降低(P < 0.01),未检测出体感和运动诱发电位。形态学观察结果显示模型组大鼠脊髓完全横断,而假手术组脊髓形态完整。结果提示实验成功构建了大鼠脊髓全横断模型。     

大鼠脊髓全横断损伤模型标准化的初步探索

目的:对比研究两种脊髓全横断模型大鼠的后肢运动功能评分和病理学特征,探索建立标准化的脊髓全横断模型。方法:成年雌性SD大鼠60只,随机分为A组(n=12)、B组(n=24)和C组(n=24),每组再随机分为术后7、14、28d3小组。以T9椎体为中心,A组行椎板切除,为假手术对照组;B、C两组行脊髓全横断,造成脊髓急性损伤模型,其中B组采用常规外科方法造模,C组采用标准化显微操作技术造模。各小组于术后7、14和28d行后肢运动功能BBB评分,然后采用过量麻醉处死大鼠,经心脏灌注取材,切片行HE染色。观测脊髓横断处瘢痕组织厚度、脊髓残端间距、空洞横径和脑脊液囊腔形成情况,计算瘢痕指数、残端间距指数和空洞指数,并对B、C两组的BBB评分及各项检测指标进行对比研究。结果:A组术前、术后BBB评分和脊髓病理无明显变化。B、C两组大鼠术后后肢完全性瘫痪,其中B组大鼠后肢功能无恢复。术后1～2周,C组大鼠开始出现后肢运动功能自发性恢复,且C组脊髓病理学检测指标值均明显低于B组,差异具有极显著性意义(P<0.01)。各组病理学观测指标与BBB评分之间无相关关系。结论:标准化脊髓全横断造模方法有利于消除个体差异,更有利于对治疗效果的量化分析和研究比较。     

急性脊髓损伤模型大鼠白细胞介素1β和核因子κB的表达

背景：在脊髓损伤后的继发性损伤过程中,白细胞介素1β参与刺激其他细胞因子和损伤介质的合成。 目的：观察白细胞介素1受体拮抗剂对急性脊髓损伤模型大鼠损伤脊髓白细胞介素1β与核因子κB表达的影响。 方法：采用改良Allen法建立SD大鼠急性脊髓损伤模型,造模后分别在损伤处敷含白细胞介素1受体拮抗剂或仅有生理盐水的明胶海绵,于脊髓损伤1,48,72 h取损伤段脊髓标本,免疫组织化学染色检测白细胞介素1β与核因子κB的表达。 结果与结论：经白细胞介素1受体拮抗剂治疗后,损伤脊髓组织白细胞介素1β和核因子κB的表达均显著降低。说明白细胞介素1受体拮抗剂可通过抑制白细胞介素1β和核因子κB的表达,减轻局部炎症反应,对急性脊髓损伤大鼠损伤段脊髓发挥保护作用。     

急性脊髓损伤模型大鼠与白细胞介素1受体拮抗剂的保护作用

背景：白细胞介素1受体拮抗剂对大鼠急性脊髓损伤后脊髓功能修复具有保护作用,但具体机制不明。 目的：观察白细胞介素1受体拮抗剂对大鼠急性脊髓损伤组织神经丝蛋白质200和胶质纤维酸性蛋白的影响。 方法：SD大鼠随机分成假手术组,生理盐水对照组和白细胞介素1受体拮抗剂治疗组。采用改良Allen氏打击法建立急性脊髓损伤大鼠模型。分别在建模后1,48和72 h获取损伤段8 mm脊髓标本。 结果和结论：免疫组织化学染色检测,白细胞介素1受体拮抗剂治疗组神经丝蛋白质200和胶质纤维酸性蛋白的表达较假手术组和生理盐水组高,差异有显著性意义(P < 0.05)。提示白细胞介素1受体拮抗剂可使急性脊髓损伤大鼠模型损伤段脊髓神经丝蛋白质200和胶质纤维酸性蛋白表达增加,对急性脊髓损伤发挥保护作用。     

辅酶Q10干预糖尿病牙周炎模型大鼠牙龈组织白细胞介素17，23的表达

BACKGROUND: Coenzyme Q10 participates in the electron transport of respiratory chain and possesses antioxidant, anti-apoptotic and anti-inflammatory properties. It has achieved good outcomes in cardiovascular disease, diabetes and cancer. Coenzyme Q10 may also have a certain application value in the fields of diabetes and periodontitis. OBJECTIVE: To observe the effect of coenzyme Q10 on the expression of interleukin-17 and interleukin-23 in gingival tissue of type 2 diabetic rats with periodontitis.  METHODS: Forty-eight healthy male Wistar rats were randomly divided into 3 groups: control, periodontitis+ diabetes+physiological saline and periodontitis+diabetes+coenzyme Q10. Rats in the control group were fed with normal diet and water. Rats in the periodontitis+diabetes+physiological saline and periodontitis+diabetes+ coenzyme Q10 groups were subjected to induction of periodontitis using the method of silk ligation and type 2 diabetes by feeding a high-fat and high-sugar diet and intraperitoneal injection of streptozotocin. After successful modeling, rats in the periodontitis+diabetes+coenzyme Q10 group were intragastrically administered coenzyme Q10 for 8 successive weeks. Rats in the periodontitis+diabetes+physiological saline group were administered equal amount of physiological saline. At the end of 2nd, 4th and 8th weeks after drug administration, four rats were randomly selected and sacrificed. The expression levels of interleukin-17 and interleukin-23 in gingival tissue were detected by immunohistochemistry. RESULTS AND CONCLUSION: At the end of 8th week, interleukin-17- and interleukin-23-positive expression in the periodontitis+diabetes+physiological saline group was significantly higher than that in the periodontitis+ diabetes+coenzyme Q10 group (P < 0.05). Coenzyme Q10 can reduce the expression levels of interleukin-17 and interleukin-23 in gingival tissue of type 2 diabetic rats with periodontitis, and alleviate periodontal tissue inflammation of type 2 diabetic rats with periodontitis.       

大鼠脊髓全横断后神经营养素-3在红核表达的变化

目的:研究大鼠脊髓全横断后神经营养素-3(NT-3)在红核表达的变化,为进一步探索脊髓损伤后的再生修复机制和神经营养因子治疗脊髓损伤提供实验依据。方法:雄性SD大鼠36只随机分为3组,脊髓横断组:大鼠脊髓在胸10~11节段之间完全横断,按存活时间不同又分为术后1、3、7及14d组;假手术组(只行椎板切除术);正常对照组。动物到达存活时间点后,应用免疫组织化学ABC法和图像分析技术检测NT-3在红核的表达。结果:对照组和实验组红核有NT-3阳性细胞。脊髓全横断后红核NT-3的表达逐渐增高,于术后7d达高峰,后缓慢下降。结论:脊髓全横断后红核对NT-3的表达增加,内源性NT-3的增加可能有利于受损的红核内神经元的存活与再生。     

Basal lamina at the site of spinal cord transection in the rat: An ultrastructural study

This electron microscopic study confirms that basal lamina (BL) begins to cap the cut end of the spinal cord 15 days after spinal cord transection. BL is first seen immediately adjacent to reactive glial cells but only when there is collagen in the nearby interstitial space. This finding suggests that collagen may provide a trigger to initiate the production of BL by reactive glia. We found no direct evidence that BL in this injury area impeded the outgrowth of regenerating neurites.     

Functional recovery of stepping in rats after a complete neonatal spinal cord transection is not due to regrowth across the lesion site

Rats receiving a complete spinal cord transection (ST) at a neonatal stage spontaneously can recover significant stepping ability, whereas minimal recovery is attained in rats transected as adults. In addition, neonatally spinal cord transected rats trained to step more readily improve their locomotor ability. We hypothesized that recovery of stepping in rats receiving a complete spinal cord transection at postnatal day 5 (P5) is attributable to changes in the lumbosacral neural circuitry and not to regeneration of axons across the lesion. As expected, stepping performance measured by several kinematics parameters was significantly better in ST (at P5) trained (treadmill stepping for 8 weeks) than age-matched non-trained spinal rats. Anterograde tracing with biotinylated dextran amine showed an absence of labeling of corticospinal or rubrospinal tract axons below the transection. Retrograde tracing with Fast Blue from the spinal cord below the transection showed no labeled neurons in the somatosensory motor cortex of the hindlimb area, red nucleus, spinal vestibular nucleus, and medullary reticular nucleus. Retrograde labeling transsynaptically via injection of pseudorabies virus (Bartha) into the soleus and tibialis anterior muscles showed no labeling in the same brain nuclei. Furthermore, re-transection of the spinal cord at or rostral to the original transection did not affect stepping ability. Combined, these results clearly indicate that there was no regeneration across the lesion after a complete spinal cord transection in neonatal rats and suggest that this is an important model to understand the higher level of locomotor recovery in rats attributable to lumbosacral mechanisms after receiving a complete ST at a neonatal compared to an adult stage.     

Localization of c-Fos protein in the rat spinal cord after carrageenan treatment

We have characterized segmental and laminar distribution patterns of Fos-immunopositive (Fos-IP) neurons in spinal cord segments L3-L6 after carrageenan treatment. A large number of Fos-IP neurons was found in the medial region of the ipsilateral dorsal horn laminae I-II at 4 and 6 h postinjection (pi). At one day pi, the number of Fos-IP neurons was decreased significantly, which correlated with suppression of inflammation in the affected hind paw. Bilaterally, Fos-IP neurons reappeared in the L3-L6 spinal cord segments at 3-4 days pi, mainly in the deep laminae LV-LVI. However, signs of inflammation had distinctly attenuated. Our data indicate a biphasic trend in Fos-IP in this experimental model of inflammation.     

Persistence of hybrid fibers in rat soleus after spinal cord transection

The effects of a chronic (up to 360 days) reduction in neuromuscular activity (defined as electrical activation and loading) on myosin heavy chain (MHC) isoform expression in the rat soleus muscle were studied. A complete mid‐thoracic (T7‐T8) spinal cord transection (ST) was used to induce a reduction in soleus muscle neuromuscular activity. Electrophoretic analyses revealed that MHC‐I was progressively decreased after ST, accounting for approx. 90% of the total soleus MHC in controls and only approx. 12% 1 year after ST. The reductions in the proportion of MHC‐I were countered by increases in MHC‐IIa and MHC‐IIx with the increase in MHC‐IIx preceding the increase in MHC‐IIa. Curiously, MHC‐IIb was expressed only at very low levels. Thus, a complete transformation from predominantly MHC‐I to MHC‐IIb did not occur. Many fibers (up to approx. 80%) contained multiple MHCs (hybrid fibers) after ST. The proportion of hybrid fibers was maintained at a high level (approx. 50%) 1 year after ST. These data suggest that: 1) a prolonged reduction in neuromuscular activity was not sufficient to induce high level MHC‐IIb expression by the soleus muscle; and 2) hybrid fibers were not simply transitional fibers. Thus, it appears that under appropriate conditions hybrid fibers may represent a “stable” fiber phenotype. Anat Rec 255:188–201, 1999. © 1999 Wiley‐Liss, Inc.     

Electromyographic assessment of damage to the injured spinal cord

The possibility of assessing the degree of damage to the spinal cord by electromyography in the early stage of treatment was demonstrated. The following indices obtained during attempts at movement are evidence of the absence of anatomical loss of continuity of the spinal cord: 1) in the case of injury at level D₅–D₁₀, absence of delay in activation of the lower segments of the long muscles of the trunk compared with their upper segments; 2) in the case of trauma at the level D₁₀-L₁, the presence of activity in the thigh and leg muscles, even if recordable only as single potentials of low amplitude; 3) whatever the level of trauma, the appearance of activity in symmetrical muscles of one lower limb during the attempt to move the opposite limb. In most cases, even if only one index is positive, the patient's motor functions will be restored by restitution.Material for this work was obtained by periodic examination of patients under treatment by Dr. A. N. Trankvillitati, Honored Physician of the RSFSR. Neurological examinations in the course of the investigation were made by Candidate of Medical Sciences L. S. Goncharova.Institute of Higher Nervous Activity and Neurophysiology, Academy of Sciences of the USSR, Moscow. (Presented by Academician of the Academy of Medical Sciences of the USSR A. A. Vishnevskii.) Translated from Byulleten' Éksperimental'noi Biologii i Meditsiny, Vol. 80, No. 10, pp. 25–29, October, 1975.     

Astrocyte transplantation for repairing the injured spinal cord

Spinal cord injury (SCI) leads to permanent deficits in neural function without effective therapies, which places a substantial burden on families and society. Astrocytes, the major glia supporting the normal function of neurons in the spinal cord, become active and form glial scars after SCI, which has long been regarded as a barrier for axon regeneration. However, recent progress has indicated the beneficial role of astrocytes in spinal repair. During the past three decades, astrocyte transpla...     

急性脊髓损伤后差异蛋白的表达

背景：在质谱分析中,任何一种同位素标记相对和绝对定量试剂标记的不同样本中的同一蛋白质表现为相同的质荷比,而在串联质谱中,信号离子表现为不同质荷比(114-121)的峰,因此可以分析得到相关蛋白质的定量信息。 目的：建立大鼠急性脊髓损伤后脊髓组织差异蛋白质谱,应用同位素标记相对和绝对定量技术联合LC-MS/MS质谱技术从分子水平来探索脊髓差异蛋白的表达情况。 方法：取8只SD大鼠,参照Allen’s等方法建立大鼠急性脊髓损伤模型,随机分为脊髓损伤后0 h组和8 h组,每组各4只。损伤后取脊髓组织,用同位素标记相对和绝对定量技术分析大鼠急性脊髓损伤后脊髓组织的差异蛋白质。 结果与结论：共鉴定到了220个差异表达的蛋白,上调的差异蛋白数是116个,下调的差异蛋白有104个。其中相关神经再生的差异蛋白12个,其中上调的有7个,下调的有5个。提示实验中检测到的多种差异蛋白及表达明显的神经生长因子可能作为急性脊髓损伤的生物标记物或可能作为临床管理监测急性脊髓损伤的损伤进程、靶向治疗及评估疗效的有力证据。     

新型脊髓完全横断缺损模型大鼠的建立

BACKGROUND: Rat models of complete spinal cord transection are common models for neural tissue engineering. After transecting the spinal cord by the previous methods, gap length of broken end cannot keep relatively uniform, so we cannot objectively evaluate effects of various treatments or tissue engineering materials. OBJECTIVE: The spinal cord transection models were established by using double edged micro scissors, and the feasibility of this new model was explored by comparing with the conventional method. METHODS: A total of 42 adult female Sprague-Dawley rats were divided randomly into group A (n=6), group B (n=18) and group C (n=18). Group A only received laminectomy. In the group B, the spinal cord was transected with a sharp-pointed knife. Knife point should touch anterior wall of spinal canal and sidewall bone surface. Complete spinal cord transection models were prepared by repeated cutting. In group C, complete spinal cord transection models were established by using self-made double edged micro scissors. RESULTS AND CONCLUSION: (1) At 1 week after model establishment, in the groups B and C, complete paralysis of the hind limbs was found, and BBB scores were similar. However, significant differences in the spacing of broken end were detected. (2) At 4 weeks after model establishment, hind limb functions could restore to different degrees in both groups, but no significant difference in BBB scores was found. (3) At 8 weeks after model establishment, significant differences in hindlimb motor function scores were detectable between both groups. Biotin glucosamine tracer display: In group B, a few labeled axon fibers were observed at the caudal side of the injured spinal cord. In group C, spinal cord was completely transected, and labeled axon fibers cannot be found at the caudal side. (4) Results suggested that the modeling method of self-made double edged micro scissors could effectively eliminate individual differences, contribute to quantitative analysis and comparative study of therapeutic effects.