黄贻泽,冯大雄,李 骏,康建平,叶 飞.Y27632对脊髓损伤微环境下新生大鼠背根节神经元轴突再生的影响[J].中国脊柱脊髓杂志,2010,20(9):765-770. |
Y27632对脊髓损伤微环境下新生大鼠背根节神经元轴突再生的影响 |
中文关键词: 脊髓损伤 背根节 轴突再生 Y27632 Rho蛋白激酶Ⅱ 大鼠 |
中文摘要: |
【摘要】 目的:探讨Rho蛋白激酶Ⅱ(Rho-associated coiled-coil-containing protein kinase Ⅱ,ROCK Ⅱ)特异性抑制剂Y-27632对脊髓损伤微环境下新生大鼠背根节(dorsal root ganglion,DRG)神经元轴突再生和延长的影响。方法:取新生(<5d)SD大鼠胸腰段DRG神经元进行原代培养、提纯和鉴定。将健康雌性SD成年大鼠15只随机分为脊髓损伤组、假手术组和正常组,每组5只;用WD法制成T9平面脊髓损伤动物模型,造模7d后取T8~T10节段脊髓制作脊髓提取液。将新生大鼠DRG神经元分为5组进行培养:A组,DRG神经元+PBS;B组,DRG神经元+正常组脊髓提取液;C组,DRG神经元+假手术组脊髓提取液;D组,DRG神经元+损伤脊髓提取液;E组,DRG神经元+损伤脊髓提取液+不同浓度(5、10、20、30、40、50μmol/L)Y27632。培养2d后观察并比较各组新生大鼠DRG神经元轴突平均长度和微管(Tubulin βⅢ)荧光表达强度。结果:A、B和C组平均神经轴突长度及轴突远端tubulin βⅢ表达强度比较无统计学差异(P>0.05);D组明显减小,与A、B和C组分别比较有统计学差异(P<0.05)。5、10μmol/L Y27632治疗组平均轴突长度以及轴突远端和生长锥tubulin βⅢ表达强度比D组有所增加(P<0.05),10μmol/L Y27632治疗组增加更明显,但小于A、B和C组(P<0.05)。20~50μmol/L Y27632治疗组平均轴突长度以及轴突远端和生长锥tubulin βⅢ表达强度三组间比较无统计学差异(P>0.05);与5~10μmol/L Y27632治疗组、D组比较明显增加(P<0.05);与A、B、C组比较平均轴突长度无统计学差异(P>0.05),平均荧光密度明显增加(P<0.05)。结论:损伤脊髓提取液能明显抑制新生大鼠DRG神经元轴突生长,导致轴突回缩。加入5~10μmol/L Y27632能促进轴突生长,20~50μmol/L Y27632更能明显促进轴突生长。 |
In vitro effect of Y27632 on dorsal root ganglion neurons axonal regeneration in neogenetic rats with spinal cord injury |
英文关键词:Spinal cord injury Dorsal root ganglia Axonal regeneration Y27632 Rho-associated coiled-coil-containing protein kinase Ⅱ Rat |
英文摘要: |
【Abstract】 Objective:To explore the in vitro effect of Rho-associated coiled-coil-containing protein kinase Ⅱ(ROCK Ⅱ) inhibitor(Y27632) on dorsal root ganglion(DRG) neurons axonal growth and extension with in neogenetic rats with spinal cord injury.Method:All thoracic-lumbar DRGs harvested from new-born Sprague-Dawley(SD) rats(<5d) were cultured,purified and indentified.15 adult female SD rats were randomly divided into three groups with each group of 5 animals.5 animals were subjected to weight-drop impact inducing T9 spinal cord injury(SCI),5 in sham operation group and 5 in normal group.T8-T10 spinal cord extracts(SCEs)were harvested in each group at 7th day after spinal cord injury.The experiment was divided into 5 groups which termed as followings:DRG neurons + phosphate-bufered saline(PBS)(group A),DRG neurons + normal SCEs(group B),DRG neurons + sham operation SCEs(group C),DRG neurons + SCI SCEs(group D) and DRG neurons + SCI SCEs +(5μmol/L,10μmol/L,20μmol/L,30μmol/L,40μmol/L,50μmol/L)Y27632(group E).The average axonal length and positive expression of tubulin βⅢ at distal end of neuronal axons were observed after 2 days′ co-culture respectively in intro.Result:There was no difference with regarding to average axonal length among group A,B and C respectively,while the axonal length in group D was significant shorter than in group A,B and C.The mean tubulin βⅢ fluorescence densities of axonal distal end in group D was significant weaker than in group A,B and C no difference existed between group A,B and C.In 5-10μmol/L Y27632 group,there were average longer axonal length and higher expression of tubulin βⅢ at axonal distal end than in group D,but less than in group A,B and C respectively.10μmol/L Y27632 group showed longer length and higher expression than 5μmol/L Y27632 group.In 20-50μmol/L Y27632 groups,average axonal length showed no difference but higher expression of tubulin βⅢ at axonal distal end existed compared with group A,B and C,which was more than 5-10μmol/L Y27632 groups and group D.There no difference among 20-50μmol/L Y27632 groups.Conclusion:The SCI SCEs inhibit DRG neurons axonal growth obviously,5~10μmol/L Y27632 could promote axon growth,while 20~50μmol/L Y27632 could promote axon growth obviously. |
投稿时间:2010-06-01 修订日期:2010-08-03 |
DOI:10.3969/j.issn.1004-406X.2010.[issue].765.5 |
基金项目:基金项目:国家自然科学基金课题(批准编号:30872602) |
|
摘要点击次数: 3026 |
全文下载次数: 3138 |
查看全文 查看/发表评论 下载PDF阅读器 |
关闭 |