| 脊髓损伤后脂质蓄积对损伤灶自发荧光强度的影响 |
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Received:December 17, 2015
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| 作者 | Author | 单位 | Unit | E-Mail |
| 郭磊 |
GUO Lei |
西安交通大学第二附属医院骨二科, 陕西 西安 710004 |
The Second Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi, China |
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| 程志坚 |
CHENG Zhi-jian |
西安交通大学第二附属医院骨二科, 陕西 西安 710004 |
The Second Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi, China |
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| 张舵 |
ZHANG Duo |
西安交通大学第二附属医院骨二科, 陕西 西安 710004 |
The Second Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi, China |
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| 常素娥 |
CHANG Su-e |
西安交通大学第二附属医院骨二科, 陕西 西安 710004 |
The Second Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi, China |
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| 贺西京 |
and HE Xi-jing |
西安交通大学第二附属医院骨二科, 陕西 西安 710004 |
The Second Department of Orthopaedics, the Second Affiliated Hospital of Xi'an Jiaotong University, Xi'an 710004, Shaanxi, China |
xijing_h@vip.tom.com |
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| 期刊信息：《中国骨伤》2016年29卷，第8期，第738-743页 |
| DOI：10.3969/j.issn.1003-0034.2016.08.013 |
| 基金项目:国家自然科学基金（编号：81571209） |
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| 目的：研究小鼠脊髓损伤后损伤灶的脂质蓄积与自发荧光强度之间的关系，探索硫酸铜能否消除脊髓损伤灶的自发荧光。方法：选取鼠龄8～12周，体重18～24 g的野生型小鼠36只，随机分为正常对照组（4只）与脊髓损伤组（32只）.脊髓损伤组于损伤后1、2、4、8周分别随机抽取8只处死，以损伤灶为中心取脊髓组织标本行冰冻切片（正常对照组取相同节段的脊髓，标本放入4%多聚甲醛溶液进行后固定），在荧光显微镜的绿色通道下观察自发荧光，进行油红O染色显示损伤灶的脂质蓄积，并分析自发荧光强度与脂质蓄积量的相关性。配制硫酸铜缓冲液处理切片以消除自发荧光，并优化硫酸铜浓度与作用时间。结果：正常脊髓组织切片未发现明显自发荧光或脂质染色，而脊髓损伤后在损伤灶出现自发荧光，并且强度随损伤后时间延长而增强。油红O染色显示损伤灶的脂质同样随伤后时间延长而蓄积，并且增长趋势与自发荧光的增强趋势呈正相关。应用硫酸铜缓冲液后，自发荧光强度显着降低，优化硫酸铜浓度与作用时间后效果更好。结论：脊髓损伤后损伤灶脂质蓄积可能在决定自发荧光的强度上起重要作用，自发荧光强度可以作为评估脂质过氧化损害的简便指标；优化的硫酸铜法能够显着消除脊髓损伤后损伤灶的自发荧光，有利于免疫荧光染色技术在脊髓损伤研究中的应用。 |
| [关键词]:脊髓损伤 脂质移动 自发荧光 硫酸铜 |
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| Effect of lipid accumulation after spinal cord injury in autofluorescence intensity of injury site |
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| Abstract:Objective: To investigate the relationship between lipid accumulation and autofluorescence intensity of injury site after spinal cord injury (SCI),and explore whether CuSO4 can eliminate autofluorescence in the injury site after SCI. Methods: Thirty-six Wild Type mice at age of 8 to 12 weeks (weight 18 to 24 g) were randomly divided into normal control group (n=4) and SCI group (n=32). Respectively,8 mice of SCI group were sacrificed randomly at 1 week,2 weeks,4 weeks and 8 weeks after injury. Frozen sections of spinal cord tissue with injury site at the center were made to observe autofluorescence under green channel of fluorescence microscope(Specimens of normal control group were taken from the same segment of the spinal cord,and fixed with 4% paraformaldehyde solution). Oil Red O staining was applied to visualize the lipid accumulation in the injury site,and correlation between lipid accumulation and autofluorescence intensity was analyzed. Furthermore,sections were incubated with CuSO4 buffer to eliminate autofluorescence,and CuSO4 concentration and incubation time was optimized. Results: No obvious autofluorescence or lipid staining was found in normal spinal cord tissue sections. By contrast,autofluorescence appeared in the injury site of spinal cord sections,and the intensity increased with passing time after injury. Oil Red O staining showed that lipid accumulated in the injury site with passing time after injury as well,and the correlation between lipid accumulation and autofluorescence intensity was positive. After incubation with CuSO4 buffer,the autofluorescence in the injury site was significantly reduced,especially after optimizing CuSO4 concentration and incubation time. Conclusion: Lipid accumulation may play an important role to determine the autofluorescence intensity of injury site after SCI,and the autofluorescence intensity can be used as a simple index for evaluating lipid peroxidation damage. Optimized method of using CuSO4 can significantly reduce the autofluorescence in the injury site after SCI,which will be beneficial to the application of immunofluorescence staining technique in the study of SCI. |
| KEYWORDS:Spinal cord injuries Lipid mobilization Autofluorescence Cupper Sulfate |
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| 引用本文，请按以下格式著录参考文献： |
| 中文格式: | 郭磊,程志坚,张舵,常素娥,贺西京.脊髓损伤后脂质蓄积对损伤灶自发荧光强度的影响[J].中国骨伤,2016,29(8):738~743 |
| 英文格式: | GUO Lei,CHENG Zhi-jian,ZHANG Duo,CHANG Su-e,and HE Xi-jing.Effect of lipid accumulation after spinal cord injury in autofluorescence intensity of injury site[J].zhongguo gu shang / China J Orthop Trauma ,2016,29(8):738~743 |
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