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在分离的豚鼠脊髓压迫模型中活性氧的增加及其抑制作用。

The increase of reactive oxygen species and their inhibition in an isolated guinea pig spinal cord compression model.

作者信息

Luo J, Li N, Robinson J P, Shi R

机构信息

Center for Paralysis Research, Department of Basic Medical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, Indiana 47907, USA.

出版信息

Spinal Cord. 2002 Dec;40(12):656-65. doi: 10.1038/sj.sc.3101363.

Abstract

STUDY DESIGN

In vitro studies using isolated guinea pig spinal cord.

OBJECTIVES

To develop an alternative model using isolated guinea pig spinal cord, which can be used to screen antioxidants for in vivo SCI treatment.

SETTING

Department of Basic Medical Sciences, Purdue University, West Lafayette, Indiana, USA.

METHODS

The compression injury was induced by a constant-displacement of 5-s compression of spinal cord using a modified forceps possessing a spacer. Reactive oxygen species (ROS) were evaluated using three distinct methods: fluorescence microscopy, lipid peroxidation assay, and flow cytometry.

RESULTS

The injury-mediated ROS increases are comparable with other in vivo studies and consistent with our previous observation using a similar injury model and measured with electrophysiological and anatomical technique. Further, ascorbic acid, hypothermia, or the combination of both significantly suppressed superoxide and lipid peroxidation. The combination treatment was the most effective when compared with ascorbic acid or hypothermia alone.

CONCLUSION

This in vitro model has the advantage of replicating some of the in vivo conditions while gaining the ability to control the experimental conditions. This in vitro model is suitable to study the mechanisms of ROS generation and degradation and can also be used to critically evaluate the effective suppressor of ROS in the contents of spinal cord traumatic injury.

摘要

研究设计

使用分离的豚鼠脊髓进行体外研究。

目的

开发一种使用分离的豚鼠脊髓的替代模型,可用于筛选用于体内脊髓损伤治疗的抗氧化剂。

地点

美国印第安纳州西拉斐特普渡大学基础医学系。

方法

使用带有间隔物的改良镊子对脊髓进行5秒恒定位移的压迫损伤来诱导损伤。使用三种不同方法评估活性氧(ROS):荧光显微镜检查、脂质过氧化测定和流式细胞术。

结果

损伤介导的ROS增加与其他体内研究相当,并且与我们之前使用类似损伤模型并通过电生理和解剖技术测量的观察结果一致。此外,抗坏血酸、低温或两者的组合可显著抑制超氧化物和脂质过氧化。与单独使用抗坏血酸或低温相比,联合治疗最有效。

结论

这种体外模型具有复制一些体内条件的优点,同时获得了控制实验条件的能力。这种体外模型适用于研究ROS产生和降解的机制,也可用于严格评估脊髓创伤性损伤内容物中ROS的有效抑制剂。

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