脊髓损伤c-fos基因的表达及调控的实验研究

本研究探讨了碱性成纤维细胞生长因子 (ｂＦＧＦ)和单唾液酸四己糖神经节甙子 (ＧＭ1)联合应用在脊髓损伤中对ｃ ｆｏｓ基因表达的调节作用 ,结果报道如下。1.材料和方法 :Ｗｉｓｔａｒ大鼠随机分组 :(1)对照组 ;(2 )脊髓损伤组 :腹腔注射生理盐水 1ｍｌ;(3)ＧＭ1组 :腹腔注射神经节苷脂ＧＭ12 0ｍｇ/ｋｇ;(4)ｂＦＧＦ组 :脊髓损伤后 30ｍｉｎ、1、2ｈ侧管注射ｂＦＧＦ2 0ｍｇ/ｋｇ到蛛网膜下腔 ;(5 )ＧＭ1加ｂＦＧＦ组 :联合给药 ,方法同ＧＭ1和ｂＦＧＦ组。采用Ｎｙｓｔｒｏｍ方法 ,作Ｔ8、9椎板切除 ,暴露脊髓 4ｃｍ× 4ｃｍ ,35克重锤压…     

神经生长因子保护受伤脊髓组织的实验研究

为了解神经生长因子（ＮＧＦ）对损伤的脊髓组织的作用，采用Ａｌｅｎ氏ＷＤ装置，以１０ｇ冲击棒自２．５ｃｍ高度下落撞击ＳＤ大鼠Ｔ８脊髓，并于蛛网膜下腔内置入导管。术后，实验组经导管注入ＮＧＦ溶液；对照组则注入生理盐水。术后４，８及２４ｈ，取脊髓损伤段标本，分别经干湿法、原子吸收光谱法测量水、钙含量。结果发现：损伤脊髓段组织钙含量明显增高，组织水肿严重；脊髓损伤后，应用ＮＧＦ可显著改善这些变化。这一实验结果证实ＮＧＦ对受伤的脊髓有明显的保护作用，其保护作用与稳定钙离子水平有关。     

碱性成纤维细胞生长因子对皮瓣存活的影响

目的观察碱性成纤维细胞生长因子（ｂＦＧＦ）对皮瓣存活的影响。方法采用Ｗｉｓｔａｒ大鼠背部任意皮瓣（６．０ｃｍ×１．５ｃｍ）模型，于原位缝合后即刻在皮瓣下滴注ｂＦＧＦ。结果ｂＦＧＦ治疗组皮瓣坏死率明显降低；组织学检查发现，ｂＦＧＦ治疗组皮瓣下肉芽组织层明显增厚，并有大量毛细血管和成纤维细胞增生；ｂＦＧＦ治疗组在皮瓣形成后第３，４和５天断蒂，其坏死率均明显低于对照组。结论局部应用ｂＦＧＦ可降低缺血皮瓣坏死率，并有利于皮瓣早期断蒂。     

成纤维细胞生长因子促进放烧复合伤创面愈合的实验研究

采用重组的碱性成纤维细胞生长因子（ｂＦＧＦ），观察其对大鼠重度放烧复合伤皮肤创面愈合的作用。结果提示：ｂＦＧＦ治疗组第６０天创面完全愈合的百分率较对照组提高２８．５％，５０％烧伤面积愈合所需的平均时间较对照组提前９天。病理学检查发现，ｂＦＧＦ可刺激成纤维细胞增殖及胶原合成，促进毛细血管增生和肉芽组织形成。在早期，ｂＦＧＦ尚可增强创面炎症反应，提高机体白细胞吞噬功能，加速脾细胞ＮＫ活性的恢复。     

碱性成纤维细胞生长因子与脊髓损伤

脊髓损伤 (ｓｐｉｎａｌｃｏｒｄｉｎｊｕｒｙ ,ＳＣＩ)是临床上常见的疾病 ,且致瘫率高 ,严重地危及人类健康。脊髓损伤与很多生长因子有关。研究表明 ,碱性成纤维细胞生长因子 (ｂａｓｉｃｆｉｂｒｏｂｌａｓｔ ｇｒｏｗｔｈｆａｃｔｏｒ ,ｂＦＧＦ)的生物学作用极其广泛 ,它在胚胎发育、血管生成、促进创伤愈合与组织修复、促进组织再生及神经系统生长发育等许多方面发挥着重要的生物学调节作用[1] 。现有关ｂＦＧＦ的研究不断深入 ,并取得令人鼓舞的成果。本文就ｂＦＧＦ在脊髓中的表达及ｂＦＧＦ对脊髓损伤的作用等方面的研究近况作一…     

神经生长因子保护脊髓背根神经节的实验研究

坐骨神经损伤后，在相应节段脊髓神经元可出现变性。为了观察外源性神经生长因子（ＮＧＦ）的保护作用而设计了本实验。方法是切断ＳＤ大鼠单侧坐骨神经，损伤局部给予外源性ＮＧＦ。借助酶组织化学及图像分析检测坐骨神经切断及应用ＮＧＦ后所引起的脊髓内抗氟化物酸性磷酸酶（ＦＲＡＰ）的活性变化，以探讨ＮＧＦ对脊髓背根神经节的保护作用。结果发现：坐骨神经切断可导致脊髓内ＦＲＡＰ含量降低，从而提示坐骨神经损伤可损害背根神经节；应用ＮＧＦ可显著减少这一酶含量的降低，从而首次从体内酶学水平证实ＮＧＦ对脊髓背根神经节有明显的保护作用。     

生长因子在大鼠前列腺组织中的表达及雄激素的调控作用

目的　观察雄激素水平改变对大鼠前列腺组织碱性成纤维细胞生长因子（ｂＦＧＦ） 和转化生长因子β１（ＴＧＦβ１）表达的影响。方法　采用分子杂交技术研究ｂＦＧＦｍＲＮＡ 及ＴＧＦβ１ｍＲＮＡ 在去势和未去势大鼠前列腺组织中表达的差异。结果　去势组ＴＧＦβ１ｍＲＮＡ 杂交斑点ＩＯＤ 均值为３１５．５,是未去势组（１７９）的１．７６ 倍;而未去势组ｂＦＧＦｍＲＮＡ 杂交斑点ＩＯＤ 均值为４４５ ．３,是去势组（２００．３） 的２．２２倍,差异均有显著性（Ｐ＜０ ．０１）。结论　雄激素水平变化可影响ｂＦＧＦｍＲＮＡ 及ＴＧＦβ１ｍＲＮＡ 的表达,ｂＦＧＦ及ＴＧＦβ１ 介导雄激素的作用。     

胚胎脊髓移植联合应用神经生长因子及尼莫地平对成鼠损 …

目的：研究胚胎脊髓移植联合应用神经生长因子（ＮＧＦ）及尼莫地平（ＮＤ）对损伤脊髓的修复作用。方法：９０只Ｗｉｓｔａｒ大鼠半切洞脊髓损伤后,随机分为３组：⑴单纯胚胎脊髓移植组（Ａ组）,⑵移植＋ＮＧＦ组（Ｂ组）,⑶移植＋ＮＧＦ＋ＮＤ组（Ｃ组）。观察组织形态学、运动诱发电位及行为学变化。结果：Ｃ组的运动诱发电位潜伏期在早期比Ａ组、Ｂ组皆有明显缩短（Ｐ〈０．０５）。以后肢运动功能的Ｔａｒｌｏｖ评分看,Ｂ组     

大剂量甲基强的松龙对大鼠急性脊髓半切损伤的早期神经保护作用

目的研究大剂量甲基强的松龙（methylprednisolone,MP）对大鼠急性脊髓半切损伤的早期神经保护作用。方法 50只SD大鼠随机取10只仅行椎板切除,作为对照组。其余40只随机分为2组,脊髓半切损伤组及MP治疗组各20只。各组动物再随机平分为2个小组,分别在脊髓半切损伤后1d、7d进行BBB法神经功能评分、脊髓组织形态学观察、神经中丝（neurofilament NF）和胶质纤维酸性蛋白（glial fibrillary acidic protein,GFAP）免疫组化测定。结果脊髓半切损伤后立即使用大剂量MP明显提高了伤后7d时MP治疗组的BBB评分;对损伤脊髓的组织形态学有明显改善;明显提高了NF的表达,抑制了GFAP的表达。结论早期大剂量使用MP对大鼠急性脊髓半切损伤有神经保护作用。     

碱性成纤维细胞生长因子在大鼠坐骨神经损伤修复中的作用

目的　研究碱性成纤维细胞生长因子（ｂ Ｆ Ｇ Ｆ）对周围神经损伤的促修复作用。方法　建立大鼠坐骨神经离断模型,镜下进行神经外膜缝合,局部滴加药物和术后肌注ｂ Ｆ Ｇ Ｆ 以及临床用药神经生长因子（ Ｎ Ｇ Ｆ）,通过神经传导速度和功能指数评定观察坐骨神经修复情况。结果　术后１ 个月,以正常对照为标准,０ 基线为正常水平,生理盐水、 Ｎ Ｇ Ｆ、ｂ Ｆ Ｇ Ｆ 使用组神经功能指数评定结果分别为：－ １１４．３０±１０．３４、－ ７０．５０±１１．０１、－ ５０．４５±７．８２;术后３ 个月,分别为：－ ５４．９６±１６．４６、－ ３５．２１±１０．８０、－ ２７．５３±１１．２３,ｂ Ｆ Ｇ Ｆ组神经传导速度明显快于生理盐水组和 Ｎ Ｇ Ｆ 组（ Ｐ＜ ０．０１）。结论　ｂ Ｆ Ｇ Ｆ对坐骨神经损伤具有显著的促功能修复作用,并且在神经损伤早期就能最有效地发挥作用,效果明显优于临床用药 Ｎ Ｇ Ｆ。     

尾静脉注射bFGF对大鼠脊髓损伤的早期影响

目的:比较不同途径应用碱性成纤维细胞生长因子(basicfibroblastgrowthfactor,bFGF)对大鼠脊髓损伤早期水、钙、镁离子的影响,探讨静脉应用bFGF的可行性。方法:AllenWD(Weightdrop)技术,以10g×25cm致伤力造成大鼠T8急性脊髓不完全损伤,不同途径用药比较。术后2h、6h及24h切取伤区脊髓组织检测水、钙、镁离子变化。结果:受损伤脊髓组织水含量增多,钙离子水平增高,镁离子水平下降,而蛛网膜下腔应用bFGF及尾静脉注射bFGF均能明显改变上述变化。结论:脊髓损伤早期静脉应用bFGF同样对脊髓损伤具有保护作用。     

Basic fibroblast growth factor (bFGF) enhances tissue sparing and functional recovery following moderate spinal cord injury.

The rapid increase in basic fibroblast growth factor (bFGF) production following spinal cord injury (SCI) in rats is thought to serve a role in the cellular processes responsible for the functional recovery often observed. In this study, bFGF was intrathecally administered continuously for 1 week beginning 30 min after a moderate (12.5 mm) spinal cord contusion in adult rats using the New York University impactor device. Osmotic minipumps were implanted into the lateral ventricle and lumbar thecal sac to deliver bFGF at a rate of 3 microg or 6 microg per day versus control vehicle. Animals were behaviorally tested for 6 weeks using the Basso, Beattie, Bresnahan locomotor rating scale and histologically assessed for both tissue sparing and glial reactivity rostral and caudal to the lesion. Rats treated with bFGF regained coordinated hindlimb movements earlier than controls and demonstrated consistent coordination from 4 to 6 weeks. Vehicle-treated rats showed only modest improvements in hindlimb function. The amount of spared tissue was significantly higher in bFGF-treated rats than in controls. Astrocyte and microglial reactivity was more pronounced in bFGF-treated animals versus controls. In summary, intrathecal infusion of exogenous bFGF following SCI significantly reduces tissue damage and enhances functional recovery. Early pharmacological intervention with bFGF following SCI may serve a neuroprotective role and/or create a proregenerative environment, possibly by modulating the neuroglial response.     

Effect of basic fibroblast growth factor on the expression of glial fibrillary acidic protein after tractive spinal cord injury in rats

Basicfibroblastgrowthfactor(bFGF) isanewneurotrophicfactor(NTF)andhasbeencloselystudiedinrecentyears.1 Itsneurotrophiceffectsincludepromotioninneuronalregenerationandsurvival. StudieshavefoundthatexogenousbFGFpumpedintothemarroworthesheathcandecreasetissu…     

Secondary degeneration reduced by inosine after spinal cord injury in rats

STUDY DESIGN: Assessment of the potential protective effects of inosine on an animal model of spinal cord injury. OBJECTIVES: Our previous studies have demonstrated that inosine can directly protect neurons in vitro from zinc-induced injury and axotomized retinal ganglion cells of rats in vivo. This investigation was carried out to examine the possible protective effects of inosine on spinal cord secondary degeneration. SETTING: Institute of Neurosciences, The Fourth Military Medical University, Xi'an, China. METHODS: Compressive spinal cord injury (95-g load for 1 min) model was established in rats, and inosine was administrated beginning at different time points (2, 12, or 24 h) after spinal cord injury. RESULTS: Using terminal deoxynucleotidyl transferase-mediated dUTP-biotin nick end labeling (TUNEL) technique and hematoxylin and eosin staining, our study demonstrated that administration of inosine as late as 12 h after injury significantly reduced the total volume of spinal cord degenerative areas and the number of apoptotic cells 3 days following the trauma. CONCLUSION: Inosine can significantly reduce the spread of secondary degeneration and the cell death following spinal cord injury in adult rats. These findings may find a clinical application in the treatment of acute spinal cord injury.     

汉防己甲素治疗急性脊髓损伤的实验研究

目的 探讨汉防己甲素(Tetrandrine，Tet)对急性脊髓损伤(Acute spinal cord injuries，ASCI)的保护作用。方法 74只Wistar大鼠随机分为4组，用改良Allen’s打击法制备大鼠截瘫模型，造模前各组分别投给相应的药物，伤后1h、4h取损伤区脊髓组织进行形态学观察和生化指标测定。结果 汉防己甲素能明显降低组织钙总量和MDA含量，防止镁总量和SOD含量的下降，稳定6-Keto-PGFla／TXB2的比值，减轻组织病损。结论 Tet对ASCI有保护作用，其作用机制为①扩张微血管，改善微循环，逆转ASCI后早期缺血倾向；②抗氧自由基，减轻组织过氧化损伤，稳定生物膜性结构；③减少ASCI后钙内流，从而阻断继发性损伤的链式反应。     

神经毒素在脊髓损伤中的作用

目的 探讨神经毒素兴奋性氨基酸（ＥＡＡ）和神经肽在脊髓损伤的作用。方法 应用高效液相色谱法检测脊髓损伤后伤段组织中谷氨酸（Ｇｌｕ）天门冬氨酸（Ａｓｐ）两种ＥＡＡ的含量变化；应用放射免疫分析技术测定脊髓损伤段组织中神经肽之一强腓肽ＤｙｎＡ１－１３含量的变化；观察了蛛网膜下腔注射Ｇｌｕ，ＤｙａｎＡ１－１３兴奋性氨基酸受体拮抗剂３－（２－羧基哌嗪）丙基－１－磷酸（ＣＰＰ），ＤｙｎＡ１－１３抗血清（Ａｎ－     

Treatment with basic fibroblast growth factor-incorporated gelatin hydrogel does not exacerbate mechanical allodynia after spinal cord contusion injury in rats

Abstract

Besides stimulating angiogenesis or cell survival, basic fibroblast growth factor (bFGF) has the potential for protecting neurons in the injured spinal cord.

Objective

To investigate the effects of a sustained-release system of bFGF from gelatin hydrogel (GH) in a rat spinal cord contusion model.

Methods

Adult female Sprague–Dawley rats were subjected to a spinal cord contusion injury at the T10 vertebral level using an IH impactor (200 kdyn). One week after contusion, GH containing bFGF (20 µg) was injected into the lesion epicenter (bFGF – GH group). The GH-only group was designated as the control. Locomotor recovery was assessed over 9 weeks by Basso, Beattie, Bresnahan rating scale, along with inclined plane and Rota-rod testing. Sensory abnormalities in the hind paws of all the rats were evaluated at 5, 7, and 9 weeks.

Results

There were no significant differences in any of the motor assessments at any time point between the bFGF – GH group and the control GH group. The control GH group showed significantly more mechanical allodynia than did the group prior to injury. In contrast, the bFGF – GH group showed no statistically significant changes of mechanical withdrawal thresholds compared with pre-injury.

Conclusion

Our findings suggest that bFGF-incorporated GH could have therapeutic potential for alleviating mechanical allodynia following spinal cord injury.     

大鼠脊髓急性损伤后神经细胞凋亡及相关基因表达△

目的研究脊髓急性损伤后神经细胞的凋亡及相关基因的表达.方法大鼠脊髓(T8、T9)经中度压迫损伤后,分别在30min、2h、4h、8h、24h、48h、72h、7d、14d和21d处死取材(各时间组n=4).应用HE染色、免疫组化及凋亡细胞原位末端标记法对脊髓组织进行标记.结果损伤4h后,在损伤段及邻近段可见末端标记阳性神经元,损伤段灰质中阳性细胞数8h达高峰,24h白质中阳性胶质细胞数量达高峰.相邻节段阳性细胞数72h达高峰.损伤后P53及Bax大量表达,而Bcl-2仅少量表达.结论脊髓损伤后神经细胞的凋亡是继发损伤期的重要病理变化.     

Decreased bone anabolic effect of basic fibroblast growth factor at fatty marrow sites in ovariectomized rats

The purpose of the study was to compare the bone anabolic effects of basic fibroblast growth factor (bFGF) at hematopoietic (red) and fatty (yellow) marrow sites in ovariectomized (ovx) rats. Female Sprague Dawley rats were subjected to ovariectomy or sham surgery at 3 months of age and maintained untreated for 2 months after surgery. Three groups of ovx rats were then injected intravenously with bFGF for 14 days at a dose of 200 microg/kg body weight. One group of bFGF-treated OVX rats was killed at the end of the treatment period, whereas the other two groups were killed at 7 or 14 days after withdrawal of bFGF treatment. Another group of ovx rats and a group of sham-operated control rats were treated intravenously with vehicle alone for 14 days. The proximal tibia and first lumbar vertebra, bone sites with hematopoietic marrow, as well as the distal tibia and caudal vertebra, bone sites with primarily fatty marrow, were processed undecalcified for quantitative bone histomorphometry. At the hematopoietic marrow sites, bFGF treatment induced a marked accumulation of osteoid, which calcified during the withdrawal period to result in a significant increase in cancellous bone volume. Osteoblast and osteoid surfaces were increased by at least a factor of 10 at these sites in bFGF-treated ovx rats before declining rapidly during the withdrawal period. In contrast, osteoid volume was negligible in the fatty marrow sites of bFGF-treated ovx rats. Although these animals exhibited a nonsignificant trend for increased cancellous bone volume in the fatty distal tibia during the withdrawal period, no such trend was observed in the fatty caudal vertebra. Indices of bone formation (osteoblast and osteoid surfaces) were significantly increased by bFGF treatment in the fatty distal tibia, which retained some small pockets of hematopoietic cells, but not to the same great extent as in the skeletal sites with hematopoietic marrow. Furthermore, not even a trend for increased osteoblast and osteoid surfaces was observed in the fatty caudal vertebra of bFGF-treated ovx rats. These findings indicate that bFGF is a strong bone anabolic agent at skeletal sites with hematopoietic marrow, but the stimulatory effects of the growth factor on bone formation are greatly attenuated at fatty marrow sites.     

NGF、BDNF基因修饰嗅鞘细胞脊髓内移植对损伤脊髓细胞的保护作用

目的：观察神经生长因子(nerve growth factor，NGF)和脑源性神经营养因子(brain-derived neurotmphic fac-tor，BDNF)基因修饰的嗅神经鞘细胞(Olfactory ensheathing cells，OECs)移植对损伤脊髓组织的保护作用。方法：将脊髓半横断伤SD大鼠模型，随机分为：NGF、BDNF基因修饰的OECs移植组(A组)、OECs移植组(B组)、损伤对照组(C组)和正常对照组(D组)。24h后每组8只动物取伤段标本，测水离子含量。其余动物第6周和12周每组8只动物爬坡试验，评价下肢运动功能及运动诱发电位(MEP)检测。结果：脊髓损伤(SCI)后组织水肿，Na^ 、Ca^2 离子浓度升高，K^ 、Mg^2 离子浓度降低。NGF、BDNF、基因修饰的OECs脊髓内移植后显著改善这些变化，且使SCI后神经功能有显著恢复。结论：NGF、BDNF基因修饰的OECs脊髓内移植对SCI有保护作用。其机制可能与减少神经细胞离子失衡，改善细胞内环境有关。