共查询到19条相似文献,搜索用时 171 毫秒
1.
腺病毒介导的 NT- 3基因在大鼠坐骨神经的表达 总被引:1,自引:0,他引:1
目的观察腺病毒介导的神经营养素-3(neurotrophin-3,NT-3)基因在大鼠坐骨神经雪旺细胞(Schwanncells,SCs)的表达。方法NT-3重组腺病毒在293细胞中培养繁殖并测定滴度后,直接注入大鼠损伤修复的坐骨神经内,不同时间点取材,采用免疫组织化学染色检测NT-3蛋白的表达,并用LEICAM550型图像分析仪对坐骨神经切片NT-3免疫组化染色强弱进行定量评价。结果坐骨神经损伤修复后直接注射Ad-NT-3,2d后出现NT-3免疫组化染色阳性产物,主要位于吻合口附近,阳性产物呈平行条纹状排列,7d时显著增加(与2d组相比,P<0.01),14d和28d时有所下降(与7d组相比,P<0.01),两者差异无显著性意义(P>0.05),但与2d组相比,仍维持在较高的水平(P<0.01)。而正常坐骨神经、损伤修复后注射Ad-LacZ或生理盐水的坐骨神经NT-3免疫组化染色结果为阴性。结论NT-3基因能通过腺病毒介导转入损伤修复后周围神经的SCs并表达NT-3蛋白,为腺病毒介导神经营养因子基因治疗促进周围神经损伤再生提供了初步的理论和实验依据。 相似文献
2.
腺病毒介导的LacZ基因在大鼠坐骨神经的表达 总被引:2,自引:0,他引:2
目的 :观察腺病毒介导的LacZ基因在大鼠坐骨神经雪旺细胞 (Schwanncells ,SCs)的表达。方法 :大鼠坐骨神经内直接注射报告基因LacZ重组腺病毒 (Ad LacZ) ,X gal组织化学染色检测LacZ基因的表达 ,采用LEICAM 5 5 0型图像分析仪对坐骨神经切片X gal染色强弱进行定量评价。结果 :将滴度为 1× 10 8PFU /ml的Ad LacZ病毒液 10 μl注入坐骨神经 2d后即可检测到LacZ的表达 ,7~ 14d表达显著增加 (与 2d组比较P <0 .0 1) ,7d组与 14d组表达量无显著差异 (P >0 .0 5 ) ,2 8d时SCs蓝染又减弱 (与 7d组和 14d组比较P <0 .0 1)。注射生理盐水的对照组X gal染色阴性。结论 :腺病毒介导的LacZ基因可转入活体动物坐骨神经内并高效表达 ,表明腺病毒介导神经营养因子等基因治疗有促进周围神经损伤再生的作用。 相似文献
3.
目的观察携带小鼠脑源性神经营养因子(BDNF)cDNA表达片段的非依赖辅助复制缺陷型重组腺病毒载体(AxCA)-BDNF转染大鼠坐骨神经后基因表达情况。方法成年大鼠随机分成A组(坐骨神经缺损+硅胶管+AxCA-BDNF原液8μl);B组(坐骨神经缺损+硅胶管+BDNF溶液8μl);C组(坐骨神经缺损+硅胶管+空白病毒稀释液8μl)三组。应用原位杂交和免疫组织化学检测方法,从BDNF mRNA和蛋白水平,进行坐骨神经损伤后BDNF基因表达的定性和半定量分析测定。结果伤后腺病毒介导的BDNF基因转移组,在3、7、14d、1个月,4个时相点,近、远端神经干和脊髓(L3-6)中BDNF mRNA水平均远远高于其他两组,BDNF的水平也远远高于作为对照的单纯硅胶管套接组。结论通过腺病毒介导转染的BDNF基因在大鼠坐骨神经SCs内得到了有效表达,并通过轴突逆行转运到了相应的脊髓神经元。 相似文献
4.
壳聚糖-胶原生物膜介导雪旺细胞联合神经干细胞桥接周围神经缺损 总被引:1,自引:0,他引:1
目的 观察以壳聚糖一胶原偶联的生物膜为载体,联合移植雪旺细胞(SCs)与神经干细胞(NSCs)在坐骨神经损伤修复过程中所起的作用,评价该方法的疗效.方法 分离纯化SCs和NSCs,传代4代后共培养于壳聚糖一胶原生物膜上.32只Wistar成鼠(体重350~400 g)建立坐骨神经缺损动物模型,随机分为4组:空白组;SCs移植组;NSCs移植组;NSCs联合SCs移植组.14周后观察Cy3荧光素染色、MBP免疫酶染色及苏木素-伊红(HE)染色的结果.结果 BDA示踪显示术后联合移植组和SCs组有较多的神经纤维穿过缺损部位,大鼠右后肢感觉和运动功能恢复较其他组明显,修复神经段神经传导速度比值也大于其他组(P<0.05).SCs组和联合移植组间差异无统计学意义(P>0.05).结论 NSCs和SCs与壳聚糖一胶原生物膜相容性良好,壳聚糖一胶原生物膜介导的SCs联合NSCs桥接周围神经缺损可使部分神经再生. 相似文献
5.
神经营养素-3对大鼠急性脊髓损伤后Bcl-2和Bax表达的影响 总被引:1,自引:1,他引:0
目的:观察神经营养素-3(NT-3)对大鼠急性脊髓损伤后B细胞淋巴瘤/白血病基因-2(Bcl-2)和B细胞淋巴瘤/白血病基因伴随蛋白x(Bax)表达的影响,探讨NT-3对脊髓损伤的作用及其可能的分子机制。方法:105只SD大鼠随机分为假手术组(A组)、损伤对照组(B组)和NT-3治疗组(C组),每组35只。B、C组用改良Allen′s法以30g·cm致伤大鼠T8脊髓制作损伤模型,C组经蛛网膜下腔导管于术后即刻、4h、8h、12h、24h、3d、7d注入NT-320μl(含NT-3200ng),B组在相同时间点给予等量生理盐水;A组打开椎板后蛛网膜下腔置管,不损伤脊髓,不给药。术后24h、3d、7d、14d对大鼠进行脊髓运动功能(BBB)评分。于术后4h、8h、12h、24h、3d、7d、14d处死动物(n=5),取T8节段脊髓,甲苯胺蓝(Nissl)染色观察脊髓前角运动神经元变化情况,用免疫组织化学方法检测Bcl-2和Bax在脊髓前角运动神经元中的表达变化情况。结果:各时间点C组和B组大鼠BBB评分均显著低于A组(P<0.01),但C组显著高于B组(P<0.05或0.01)。Nissl染色C组大鼠脊髓损伤区较B组出血少、残存神经元多,A组正常。各时间点C组和B组大鼠损伤脊髓前角运动神经元中Bax阳性细胞平均光密度(AOD)值均显著高于A组(P<0.01),但C组显著低于B组(P<0.05或0.01);各时间点C组和B组大鼠损伤脊髓前角运动神经元中Bcl-2阳性细胞AOD值均显著低于A组(P<0.01),但C组显著高于B组(P<0.05或0.01)。结论:NT-3可能通过抑制Bax表达,提高Bcl-2表达,抑制脊髓损伤后神经元凋亡,从而保护损伤的脊髓组织,这可能是NT-3对脊髓损伤具有保护作用的机制之一。 相似文献
6.
目的评价将人脐血间充质干细胞(human umbilical cord blood mesenchymal stem cells,h UCBMSCs)来源的类雪旺细胞(Schwann cells,SCs)作为种子细胞,修复大鼠坐骨神经15 mm缺损的效果,为h UCBMSCs应用于临床治疗周围神经缺损提供实验依据。方法 SPF级3月龄雄性SD大鼠45只,体重200~250 g。取新生儿脐带血,淋巴细胞分离液复合高分子量羟乙基淀粉分离培养h UCBMSCs并鉴定;取第3代h UCBMSCs,采用改良化学诱导法联合细胞因子方法诱导分化培养类SCs并鉴定。取15只SD大鼠坐骨神经,采用液氮反复冻融振荡洗涤法制备去细胞神经基膜管作为支架材料;将密度1×107个/m L的类SCs细胞悬液多点注射至该支架内复合培养7 d构建组织工程神经。取SD大鼠30只制备长约15 mm坐骨神经缺损动物模型,根据缺损神经修复方法不同,实验分为A、B、C 3组(n=10),A组采用组织工程神经缝合,B组采用未复合类SCs的去细胞神经基膜管缝合,C组采用自体坐骨神经原位缝合。术后行大体观察、坐骨神经功能指数(sciatic function index,SFI)测定、神经电生理功能检测、腓肠肌湿重测定、Masson染色评价神经修复情况。结果分离培养的h UCBMSCs高表达MSCs表面标志;诱导培养后类SCs经免疫细胞化学染色检测示神经胶质细胞标志物S100b、胶质纤维酸性蛋白、P75表达呈阳性。术后8周,大体观察示A组组织工程神经管壁无坏死及液化,周围轻度粘连,吻合处连续性较好;B组支架外观与A组相似;C组自体神经周围粘连较A、B组轻,吻合口光滑,无明显膨大,颜色与正常神经相似。各组大鼠术后SFI随时间延长呈逐渐降低趋势,C组SFI恢复优于A、B组,A组优于B组(P0.05)。术后各组大鼠远端吻合口处均可检测到神经复合动作电位,波幅及传导速度C组均优于A、B组,A组优于B组(P0.05)。术后各组大鼠实验侧小腿腓肠肌与健侧相比,均发生不同程度萎缩;腓肠肌湿重恢复率C组优于A、B组,A组优于B组(P0.05)。Masson染色示A组可见大量再生神经纤维,有髓神经纤维排列较为整齐、致密,纤维直径相似;C组有髓神经纤维密度、直径及髓鞘厚度和轴突直径均明显多于A、B组,A组多于B组(P0.05)。结论 h UCBMSCs来源的类SCs能够促进大鼠15 mm坐骨神经损伤修复可作为组织工程神经种子细胞来源。 相似文献
7.
《中国修复重建外科杂志》2018,(12)
目的探讨脂肪干细胞(adipose-derived stem cells,ADSCs)来源外泌体对周围神经损伤后再生的影响,为周围神经损伤寻找新的治疗方法。方法将36只成年SD大鼠(雌雄不限,体质量220~240 g)随机分成3组,每组12只。A组为正常对照组,B组为坐骨神经挤压损伤组,C组为ADSCs来源外泌体治疗坐骨神经挤压损伤组。A组仅暴露坐骨神经后直接缝合切口,B、C组制备坐骨神经挤压损伤模型;术后次日开始,A、B组于大鼠尾静脉注射PBS液200μL,C组注射含100μg ADSCs来源外泌体的PBS液200μL,每周注射1次,连续12周。注射结束1周后处死大鼠,取损伤处坐骨神经,分别行大体观察、HE染色观察神经束情况、TUNEL检测坐骨神经雪旺细胞(Schwann cells,SCs)凋亡情况,透射电镜观察坐骨神经超微结构和SCs自噬情况。结果大体观察示A组患肢无明显异常,B、C组患肢瘫痪、肌肉萎缩,但C组瘫痪及肌肉萎缩程度轻于B组。HE染色示,A组神经束膜形态规则;B组神经束形态破坏、束膜不规则,有较多无细胞结构和组织碎片;C组神经束膜较完整,明显优于B组。TUNEL检测示,B、C组SCs凋亡细胞数显著多于A组,B组显著多于C组,差异均有统计学意义(P0.01)。透射电镜观察示,B、C组SCs自噬体较A组明显增加,但C组少于B组。结论 ADSCs来源外泌体对周围神经损伤后再生有一定促进作用,其机制可能与减少SCs凋亡、抑制其自噬、减轻神经瓦勒变性有关。 相似文献
8.
目的观察携带小鼠BDNFcDNA表达片段的重组腺病毒载体AxCA-BDNF转染大鼠坐骨神经后基因表达情况。方法成年大鼠随机分成A组(坐骨神经缺损加硅胶管加AxCA-BDNF原液8μl);B组(坐骨神经缺损加硅胶管加BDNF溶液8μl)和C组(坐骨神经缺损加硅胶管加空白病毒稀释液8μl)等三组。应用原位杂交和免疫组化等手段,从BDNFmRNA和蛋白水平,进行坐骨神经损伤后BDNF基因表达的定性和半定量分析测定。结果伤后腺病毒介导的BDNF基因转移组,在3、7、14d和1个月4个时相点,近、远端神经干和脊神经(L3-6)中BDNFmRNA水平均远远高于其它两组,BDNF的水平也远远高于作为对照的单纯硅胶管套接组。结论通过腺病毒介导转染的BDNF基因在大鼠坐骨神经许旺细胞内得到了有效表达,并通过轴突逆行转运到了相应的脊髓神经元。 相似文献
9.
《中国修复重建外科杂志》2016,(8)
目的构建携带NGF及髓磷脂相关糖蛋白(myelin associated glycoprotein,MAG)基因序列的双基因共表达腺病毒(adenovirus expressing NGF and MAG,Ad-NGF-MAG),探讨其在大鼠坐骨神经损伤修复中的作用。方法将NGF和MAG共同克隆至5型腺病毒穿梭质粒p CA 13,并在HEK 293细胞中包装得到重组腺病毒Ad-NGF-MAG并测序鉴定。取雄性SD大鼠32只,体质量180~200 g;随机分成4组(n=8):对照组(正常对照)、空病毒组(Ad组)、单独表达NGF组(Ad-NGF组)和共表达NGF、MAG组(Ad-NGF-MAG组)。Ad组、AdNGF组和Ad-NGF-MAG组大鼠制备右侧坐骨神经损伤模型后,分别于术侧腓肠肌注射空腺病毒、Ad-NGF及AdNGF-MAG(1×108 PFU),隔天1次,共3次。对照组仅暴露右侧坐骨神经后关闭切口,术后对应时间点注射生理盐水10μL。术后31 d,分别行坐骨神经功能指数(sciatic nerve function index,SFI)、神经电生理检测;切取坐骨神经标本,行RT-PCR及Western blot检测NGF及MAG m RNA及蛋白表达水平,以及组织学观察。结果实验成功构建重组腺病毒Ad-NGF和Ad-NGF-MAG。32只大鼠术后均成活,切口Ⅰ期愈合。Ad-NGF-MAG组SFI、神经传导速度、诱发电位波幅、潜伏期均明显优于Ad-NGF组和Ad组,但未达对照组水平,比较差异均有统计学意义(P0.05)。Ad-NGF-MAG组内MAG m RNA和蛋白表达最高,NGF m RNA和蛋白表达高于对照组和Ad组,比较差异均有统计学意义(P0.05)。组织学观察显示,对照组神经连续性好,Ad组神经纤维层次混乱,Ad-NGF组神经纤维层次结构清晰,局部断端再生良好,但神经纤维结构紊乱;Ad-NGF-MAG组神经纤维生长有序,神经直径较Ad-NGF组粗,神经纤维结构良好。结论坐骨神经损伤后修复过程中,腺病毒介导NGF与MAG共表达,既可促进神经纤维生长,又可抑制神经异常分支形成,促进神经结构与功能的恢复。 相似文献
10.
腺病毒介导的NT-3基因在骨髓间质干细胞中的表达 总被引:1,自引:0,他引:1
目的:研究腺病毒介导的NT-3基因在培养的大鼠骨髓间质干细胞(mesenchymal stem cells,MSCs)中的表达.方法:在293细胞中培养扩增NT-3重组腺病毒(adenovirus vector for NT-3,Ad-NT-3),测定病毒滴度,然后用Ad-NT-3感染传代培养的MSCs,RT-PCR技术检测NT-3基因的表达.结果:Ad-NT-3扩增后获得了较高滴度的病毒,MSCs经Ad-NT-3感染后有NT-3 mRNA的转录.结论:腺病毒介导的NT-3基因可转入培养的MSCs并高效表达,为NT-3基因治疗的研究奠定了基础. 相似文献
11.
Expression of adenovirus-mediated neurotrophin-3 gene in Schwann cells of sciatic nerve in rats 总被引:6,自引:0,他引:6
Successfulperipheralnerveregenerationaftermicrosurgicalrepairrequiresoptimalconditionsinthemicro milieu .Neurotrophicfactors(NTFs) ,mainpartsoftheregenerativemicro milieu ,havebeenshowntoplayanessentialtrophicroleinthedevelopmentandregenerationofperiphera… 相似文献
12.
小间隙桥接周围神经对缝合口神经调节蛋白-1表达变化的影响 总被引:2,自引:1,他引:1
目的 观察采用小间隙桥接法修复大鼠坐骨神经损伤时套管内神经调节蛋白-1(NRG-1)mRNA含量的变化.方法 取SD大鼠78只,体质量200~250 g.随机分为3组,其中实验组A和实验组B各有大鼠36只,正常对照组有大鼠6只.2组实验组根据观察时间的不同再分为6组,每组有大鼠6只.实验组A:切断并原位缝合右侧坐骨神经.实验组B:切断并采用小间隙桥接法修复右侧坐骨神经.分别于术后1、3、5、7、14、28 d以缝合口为中心,取上下各5 m共1 cm坐骨神经提取总RNA,采取实时荧光定量反转录聚合酶联(Real-time RT-PCR)技术检测组织中NRG-1mRNA的含量变化.结果 大鼠周围神经中NRG-1 mRNA含量在正常组织有低水平表达,在损伤后1 d即明显升高,在损伤后14 d降低,随即恢复较高水平,持续至伤后28 d.在部分时间点高于外膜缝合组.结论 大鼠坐骨神经损伤后采用不同修复方法NRG-1基因表达变化不同步,提示套管形成的封闭空间蕴含了较高的神经再生相关因子,有利于神经再生. 相似文献
13.
Remote adenoviral gene delivery to the spinal cord: contralateral delivery and reinjection 总被引:5,自引:0,他引:5
OBJECTIVE: This study characterizes the distribution of adenoviral genes in the spinal cord after viral vector injection into the sciatic nerve. It also evaluates the ability of repeated adenoviral sciatic nerve injections to prolong gene expression in the spinal cord. METHODS: Rat sciatic nerves were unilaterally coinjected with the retrograde tracer Fluoro-Gold (Fluorochrome, Inc., Denver, CO) and the adenoviral vector Ad5RSVntLacZ. The distribution of adenoviral gene expression in the spinal cord was compared with that of Fluoro-Gold. Next, levels of gene expression in the sciatic nerve and spinal cord were compared after single and repeated injections of Ad5RSVntLacZ. Finally, remote spinal cord gene expression in naive animals was compared with expression in animals that had been pretreated with subcutaneous Ad5RSVntLacZ inoculation. RESULTS: Viral gene expression was detected in all quadrants of the spinal cord gray matter, whereas Fluoro-Gold was detected only in the ipsilateral ventral horn (n = 5). This remote delivery was blocked by sciatic nerve transection (n = 10). Viral gene expression occurred in the sciatic nerve after both initial and repeated injections, whereas remote gene expression in the spinal cord was observed only after primary sciatic nerve injection (n = 24; P < 0.003). As with repeated sciatic nerve injections, subcutaneous inoculation with Ad5RSVntLacZ blocked subsequent remote spinal cord gene delivery (n = 8; P < 0.05). CONCLUSION: Remote viral gene delivery occurs in neurons without direct sciatic nerve projections but is dependent on intact peripheral nerves. Repeated injections fail to boost spinal cord gene expression, because of immune recognition of reinjected virus. 相似文献
14.
Intraneural colchicine inhibition of adenoviral and adeno-associated viral vector remote spinal cord gene delivery 总被引:4,自引:0,他引:4
Boulis NM Willmarth NE Song DK Feldman EL Imperiale MJ 《Neurosurgery》2003,52(2):381-7; discussion 387
OBJECTIVE: The mechanism of remote viral gene delivery to the spinal cord is unknown. The present experiment demonstrates that intraneural injection of colchicine is capable of inhibiting remote delivery of both adenoviral and adeno-associated viral (AAV) vectors, implicating axonal transport in this process. METHODS: The right sciatic nerves of adult Sprague-Dawley rats were injected with phosphate-buffered saline (PBS) (n = 5) or 10 (n = 7) or 100 (n = 4) microg colchicine. Two days later, the nerves of all animals were initially injected with 1.2 x 10(7) plaque-forming units of Ad5RSVntLac-Z. Two separate groups were injected concurrently with vector and PBS (n = 5) or 10 microg colchicine (n = 5). In a second experiment, the right sciatic nerves of CD1 mice were preinjected with PBS (n = 6) or 10 microg colchicine (n = 5). Two days later, the nerves were injected with rAAVCAG-EGFPwpre (an adeno-associated vector carrying the green fluorescent protein gene). In both experiments, sciatic nerves and spinal cords were removed and analyzed for gene expression. RESULTS: Sciatic nerve vector injection resulted in expression in both the nerve injection site and neuronal cell bodies located predominantly in the ipsilateral ventral horn. Analysis of variance revealed a significant treatment effect for 10 and 100 microg intraneural colchicine with inhibition of remote adenoviral delivery at 10 microg and blockade of remote delivery at 100 microg (P < 0.001). Colchicine injection concurrent with and before vector injection had similar inhibitory effects. Two-way analysis of variance revealed significant colchicine inhibition of remote delivery in both adenovirus- and AAV-injected animals (P < 0.003) but no dose-by-vector interaction, suggesting that both vectors are equally inhibited by colchicine. CONCLUSION: Colchicine inhibits remote spinal cord delivery of adeno-associated and adenoviral vectors in a dose-dependent manner, suggesting that remote delivery is dependent on retrograde axonal transport. 相似文献
15.
G M Rayan S I Said S L Cahill J Duke 《The Journal of hand surgery, European volume》1991,16(5):515-518
The role of vasoactive intestinal peptide (V.I.P.) in nerve regeneration was investigated by assessing the changes in immunoreactive V.I.P. levels in rat sciatic nerves following injury and repair. 60 rats were divided into three surgical groups and one control group: In group I (primary repair), sciatic nerves were divided and immediately repaired; in group II (secondary repair), sciatic nerves were divided and repaired two weeks later; in group III (no repair), sciatic nerves were divided and not repaired; and in group IV (controls), sciatic nerves were exposed but not divided. Animals were sacrificed at three days and at weekly intervals. Their sciatic nerves were extracted and assayed for V.I.P. concentrations by a specific radioimmunoassay. The mean V.I.P. concentration varied between 22 and 46 pg./mg. protein in the control nerves and between 60 and 529 pg./mg. protein in all other groups. In the three surgical groups the levels were significantly higher in proximal than in distal stumps. Following nerve injury, there was an increase in V.I.P. concentration in the injured and repaired areas. This increase was greater in injured non-repaired areas and was highest in the first 48 hours, but continued during regeneration. The accumulation of V.I.P. in divided nerves occurred in response to nerve injury. 相似文献
16.
Adenoviral gene transfer in the peripheral nervous system 总被引:6,自引:0,他引:6
Tomoko Saito Watanabe Seiji Ohtori Masao Koda Yasuchika Aoki Hideo Doya Hiroshi Shirasawa Masashi Yamazaki Hideshige Moriya Kazuhisa Takahashi Toshihide Yamashita 《Journal of orthopaedic science》2006,11(1):64-69
Background Viral vectors have gained widespread use as vehicles for somatic gene transfer, and the targeted expression of foreign proteins
by these vectors offers advantages over the systemic administration of the drugs in some therapeutic situations. Selective
virus-mediated gene transfer to the peripheral nervous system (PNS), however, remains to be established. There are no data
showing efficiency of protein transduction in the PNS, which consists of a variety of cell types, many of which are postmitotic.
Methods We prepared the first-generation replication-deficient recombinant adenovirus vectors engineered to express LacZ. Eight-week-old Wister rats were used in this study. Adenovirus vector (5 μl) containing the LacZ gene (5 × 108 pfu) was injected into rat sciatic nerves or the dorsal root ganglia at the level of L5. The sciatic nerves, the dorsal root
ganglia, and the spinal cords were obtained 7, 14, 21, and 28 days after injection. Expression of LacZ was assessed by X-gal histochemistry and β-gal immunohistochemistry.
Results Following injection of the adenovirus carrying the LacZ gene into the sciatic nerve, LacZ expression was seen mainly in the Schwann cells and the small neurons in the dorsal root ganglion. In contrast, expression
was observed in the primary nerve terminals of the spinal dorsal horn and the small to large dorsal root ganglion neurons
and the Schwann cells after injection of the vectors into the L5 dorsal root ganglion. There were no side effects in rats
with injection in the dorsal root ganglia or the sciatic nerve.
Conclusions The present study shows efficient protein transduction by adenovirus vectors in the PNS. It is noted that injection of the
virus into the dorsal root ganglia leads to extensive expression of LacZ in the spinal cord, the dorsal root ganglia, and the sciatic nerves. 相似文献
17.
目的 研究化学去细胞异体神经复合人肝细胞生长因子(HGF)修复周围神经缺损的作用.方法 体外试验检测携带人肝细胞生长因子基冈的重组腺病毒(Ad-HGF)对小鼠骨骼肌细胞的转染效率以及转染细胞对目的 蛋白的表达.化学玄细胞异体神经修复大鼠坐骨神经10mm缺损后,近、远端吻合口附近肌肉分别注射腺病毒介导的人肝细胞生长因子(Ad-HGF),与自体神经移植组对照,通过步态分析、肌肉湿重测定、轴突生长速率测定、神经电生理、计算机图像分析等指标评价神经移植后再生效果.结果 流式细胞仪结果表明,随着病毒滴度的增加,Ad-HGF对骨骼肌细胞的转染不断增加.骨骼肌细胞对目的 蛋白(HGF)的表达可以持续两周.大鼠动物实验术后16周,去细胞异体神经可以修复周罔神经缺损,同自体神经移植对比,肝细胞生长因子明显增强了去细胞异体神经的神经再生能力.结论 复合肝细胞生K因子的化学去细胞异体神经能促进神经轴突牛长速度,显著增加移植物内新生血管,满意修复一定长度周围神经缺损,可以成为一种有效的周围神经组织工程修复材料. 相似文献
18.
Kohzou Fujisawa Hitoshi Hirata Hitoshi Inada Akimasa Morita Hiroshige Hibasami 《Microsurgery》1995,16(10):684-691
The purpose of this experimental study was to investigate the usefulness and mechanism of the expansion of wallerian degenerating nerve. The study consisted of two experiments: Experiment I, functional and morphometrical analysis, and Experiment II, immunohistochemical analysis. In Experiment I, the rat nerve crush model was used to assess the effects of mechanical expansion of wallerian degenerating nerves on axonal regeneration. In Experiment II, the rat sciatic nerve cut model was used to investigate the effects of nerve expansion on Schwann cell events in wallerian degenerating nerves. In both experiments, nerve expansion was carried out between days 5 and 9 after nerve injury, using a rubber tissue expander placed beneath the sciatic nerve. In Experiment I, rats were divided into the following three groups according to the volume of saline injected: control group (nerves were crushed, without saline injection); 8-ml injection group; and 11-ml injection group. Functional recovery was assessed using the sciatic functional index until 54 days after nerve injury. Rats in all three groups showed good functional recovery, and the morphometrical analysis revealed no significant differences among the three groups. In Experiment II, anti-S-100 protein polyclonal antibody and anti-proliferating cell nuclear antigen monoclonal antibody were used to identify proliferating Schwann cells. Rats were divided into two groups: control group (nerves were cut, without expansion) and nerve expansion group. In the control group, proliferating Schwann cells were observed only between days 3 and 7. By contrast, these cells continued to be seen in the expanded nerves until day 16. These results suggest that the expansion of wallerian degenerating nerve does not have a deleterious effect on the axon-promoting property of Schwann cell tubes and that the expansion is dependent not only on the viscoelasticity of the nerves but also on enhanced proliferation of Schwann cells. © 1995 Wiley-Liss, Inc. 相似文献
19.
纤维蛋白胶粘合修复周围神经抗牵拉强度动态变化的实验研究 总被引:2,自引:0,他引:2
目的 了解用纤维蛋白胶粘合修复周围神经后早期的抗牵拉强度及其动态变化。方法 Wistar大鼠 96只 ,按手术先后随机分为神经缝合组 (n =48)和粘合组 (n =48)。切断大鼠左侧坐骨神经 ,缝合组用 11 0无创尼龙缝线作端端缝合 ,粘合组用纤维蛋白胶粘合两断端。于术后 0、3、7、14、2 1和2 8d 6个时间组 (每组n =8)取材 ,测量神经断裂时的最大负荷、功耗 ,并描绘出神经应力 -应变曲线。结果 神经抗牵拉强度曲线表现出粘弹性特性。缝合组与粘合组的最大抗牵拉强度及功耗 ,在术后 0、14、2 1和 2 8d时差异无显著意义 (P >0 .0 5 ) ;但在术后 3、7d时差异有显著意义 (P <0 .0 1,P <0 .0 5 )。结论 纤维蛋白胶有足够的抗牵拉强度 ,可以满足大鼠神经修复的需要。 相似文献