中国组织工程研究 ›› 2018, Vol. 22 ›› Issue (8): 1235-1240.doi: 10.3969/j.issn.2095-4344.0142

• 组织构建实验造模 experimental modeling in tissue construction • 上一篇    下一篇

依达拉奉干预脊髓损伤模型大鼠Ⅰ型和Ⅳ型胶原蛋白的表达

李仁斌,余光书,林焱斌,周家烽,黄奕祺,郑  伟   

  1. 厦门大学附属福州第二医院骨科,福建省福州市  350007
  • 收稿日期:2017-11-15 出版日期:2018-03-18 发布日期:2018-03-18
  • 通讯作者: 李仁斌,硕士,副主任医师,厦门大学附属福州第二医院骨科,福建省福州市 350007
  • 作者简介:李仁斌,男,1974年生,福建省明溪县人,汉族,2004年福建医科大学毕业,硕士,副主任医师,主要从事创伤骨科研究。
  • 基金资助:

    福州市科技局社会发展项目(2016-S-123-16);福建省卫生计生中青年骨干人才培养项目(2017-ZQN-73)

Edaravone effects on the expression levels of collagen type I and IV after spinal cord injury in rats

Li Ren-bin, Yu Guang-shu, Lin Yan-bin, Zhou Jia-feng, Huang Yi-qi, Zheng Wei   

  1. Department of Orthopedics, Fuzhou Second Hospital Affiliated to Xiamen University, Fuzhou 350007, Fujian Province, China
  • Received:2017-11-15 Online:2018-03-18 Published:2018-03-18
  • Contact: Li Ren-bin, Department of Orthopedics, Fuzhou Second Hospital Affiliated to Xiamen University, Fuzhou 350007, Fujian Province, China
  • About author:Li Ren-bin, Master, Associate chief physician, Department of Orthopedics, Fuzhou Second Hospital Affiliated to Xiamen University, Fuzhou 350007, Fujian Province, China
  • Supported by:

    the Social Development Project of Fuzhou Science and Technology Bureau, No. 2016-S-123-16; the Young and Middle-Aged Talent Training Program of Health and Family Planning Department of Fujian Province, No. 2017-ZQN-73

摘要:

文章快速阅读:

文题释义:
依达拉奉:是一种脑保护剂(自由基清除剂)。临床前研究提示,大鼠在缺血/缺血再灌注后静脉给予依达拉奉,可阻止脑水肿和脑梗死的进展,并缓解所伴随的神经症状,抑制迟发性神经元死亡。机制研究提示,依达拉奉可清除自由基,抑制脂质过氧化,从而抑制脑细胞、血管内皮细胞、神经细胞的氧化损伤。
脊髓功能Basso,Beattie,Bresnahan(BBB)评分:常用的运动功能评分法,此法几乎包括了脊髓损伤后运动功能恢复过程中所有行为变化,且与脊髓损伤的程度高度相符,又无需特殊设备,但标准较复杂,需对观察人员进行一定的训练,以减少主观因素的影响。操作方法是将动物放入开口盆,轻敲盆壁,使其爬行,观察动物的臀、膝、踝关节行走、躯干运动及其协调情况。总分21分。
摘要
背景:
依达拉奉是一种有效的氧自由基清除剂,有研究报道其可以有效改善脊髓损伤后肢体运动功能,但是其对损伤组织修复的作用机制尚不明确。
目的:通过观察脊髓损伤后大鼠肢体功能评分的变化及Ⅰ型胶原蛋白和Ⅳ型胶原蛋白表达情况,探讨依达拉奉促进脊髓损伤大鼠肢体功能恢复的机制。
方法:将36只大鼠随机分为3组,每组12只:假手术组不损伤脊髓,仅去除椎板后缝合+生理盐水腹腔注射;模型组去除椎板后行NYU脊髓打击器建立脊髓损伤模型+腹腔注射生理盐水;依达拉奉组:去除椎板后行NYU脊髓打击器建立脊髓损伤模型+腹腔注射依达拉奉。造模后第1天开始进行药物干预,持续7 d。干预后1,3,5及7 d对各组大鼠进行Basso Beattie Bresnahan(BBB)肢体功能评分;干预后第7天,处死大鼠,取出脊髓,尼式染色观察脊髓神经元细胞的形态;免疫组化法和Western blot检测Ⅰ型和Ⅳ型胶原蛋白表达情况。
结果与结论:①与模型组相比,干预第5天后,依达拉奉组的BBB评分显著升高(P < 0.05);②尼式染色显示假手术组脊髓灰白质界限清晰,神经细胞胞质内可见较大且明显的核仁;依达拉奉组,细胞形态较为完整,细胞水肿较轻,瘀血面积较小;模型组神经元细胞空洞较多且大,细胞核肿胀,甚至出现核仁消失的情况;③Western blot和免疫组化实验结果显示依达拉奉组的Ⅰ型胶原蛋白和Ⅳ型胶原蛋白表达水平显著高于模型组多(P < 0.05);④结果提示,依达拉奉对脊髓损伤后大鼠的肢体功能障碍有显著改善作用,可能与增加Ⅰ型胶原蛋白和Ⅳ型胶原蛋白的表达有关。

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程
ORCID: 0000-0002-2624-0497(李仁斌)

关键词: 脊髓损伤, 神经修复, 依达拉奉, 肢体功能评分, Ⅰ型胶原蛋白, Ⅳ型胶原蛋白, NYU脊髓打击器, 免疫组化, 尼式染色, Western blot法

Abstract:

BACKGROUND: Edaravone, an effective free radical scavenger, has been reported to significantly improve the rehabilitation of limb locomotion after spinal cord injury (SCI), but the underlying mechanism remains unclear.
OBJECTIVE: To explore the mechanism underlying edaravone promoting the recovery of limb locomotion in rats with SCI by observing the Basso, Beattie, Bresnahan scores and expression levels of collagen type I and IV.
METHODS: Thirty-six rats were randomly allocated into three groups (n=12 per group): sham group (laminectomy plus intraperitoneal injection of normal saline), model group (SCI model by NYU impactor plus intraperitoneal injection of normal saline), and edaravone group (SCI model by NYU impactor plus intraperitoneal injection of edaravone). All rats were given the administration at the 1st day post-SCI for consecutive 7 days. The Basso, Beattie, Bresnahan scores were tested at 1, 3, 5 and 7 days post treatment. On day 7, all rats were sacrificed to remove the spinal cord, and the morphology of neurons in the spinal cord were observed by Nissl staining; the expression levels of collagen type I and IV were detected by immunohistochemistry and western blot assays.
RESULTS AND CONCLUSION: Compared with the model group, the Basso, Beattie, Bresnahan scores in the edaravone group were significantly increased at day 5 post treatment (P < 0.05). Nissl staining showed a clear boundary between grey matter and white matter, and a large nucleolus in the neurocytoplasm in the sham group; there was a complete structure of neurons, slight cellular swelling and small hematoma area in the edaravone group; many and large cavitations and swollen nucleus were found in the neurons, even without nucleolus. Immunohistochemistry and western blot assay results showed that the expression levels of collagen type I and IV in the edaravone group were significantly higher than those in the model group (P < 0.05). These results indicate that edaravone can promote the recovery of limb locomotion of rats with SCI, probably via up-regulating the expression levels of collagen type I and IV.

中国组织工程研究杂志出版内容重点:组织构建;骨细胞;软骨细胞;细胞培养;成纤维细胞;血管内皮细胞;骨质疏松组织工程

Key words: Spinal Cord Injuries, Models, Animal, Collagen Type I, Tissue Engineering

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