天津医药 ›› 2015, Vol. 43 ›› Issue (11): 1271-1274.doi: 10.11958/j.issn.0253-9896.2015.11.013

• 实验研究 • 上一篇    下一篇

硫化氢对大鼠急性脊髓损伤的保护作用

孙平, 范仲凯, 李昊天, 王继权, 赵兴长, 李刚, 吕刚△#br#   

  1. 辽宁医学院附属第一医院骨科 (邮编 121001
  • 收稿日期:2015-04-13 修回日期:2015-07-03 出版日期:2015-11-15 发布日期:2015-11-15
  • 通讯作者: 吕刚 E-mail:ganglv_jz@163.com E-mail:ganglv_jz@163.com
  • 作者简介:孙平 (1989), 男, 硕士在读, 主要从事脊髓损伤研究
  • 基金资助:
    国家自然科学基金资助项目 (81272074); 辽宁省高等学校优秀人才支持计划项目 (LJQ2014091); 辽宁医学院校长基金 (QM2014010

Protective effects of hydrogen sulfide on acute spinal cord injury in rats

SUN Ping , FAN Zhongkai , LI Haotian, WANG Jiquan, ZHAO Xingchang, LI Gang, LYU Gang   

  1. Department of Orthopedics, First Affiliated Hospital of Liaoning Medical College, Jinzhou 121001, China
  • Received:2015-04-13 Revised:2015-07-03 Published:2015-11-15 Online:2015-11-15
  • Contact: LYU Gang E-mail:ganglv_jz@163.com E-mail:ganglv_jz@163.com

摘要: 目的 探讨硫化氢 (H2S) 对大鼠急性脊髓损伤后自噬及细胞凋亡的影响。方法 36 只健康雄性 SD 大鼠随机分为假手术组 (Sham 组)、 单纯脊髓损伤组 (模型组)、 H2S 处理组(H2S ), 每组 12 只。采用 Allens 法构建大鼠脊髓损伤模型, Sham 组只行椎板切除术, 暴露脊髓不打击; H2S 组在脊髓损伤后 1 h 腹腔注射 50 μmol/kg 硫氢化钠;模型组给予等体积生理盐水。各组脊髓损伤 24 h 后取出脊髓组织, Western blot 检测 LC3p70S6K Cleaved caspase-3 表达; 免疫荧光法检测 LC3 表达; TUNEL 染色法检测各组细胞凋亡率。结果 Sham 组相比, 模型组 LC3Ⅱ/LC3ⅠCleaved caspase-3 表达升高,p70S6K 表达降低、 细胞凋亡率升高 (P < 0.01); 与模型组相比, H2S LC3Ⅱ/LC3ⅠCleaved caspase-3 表达降低, p70S6K 表达升高, 细胞凋亡率下降(P < 0.01)。结论 H2S 可抑制大鼠急性脊髓损伤后自噬并减少细胞凋亡。

关键词: 急性脊髓损伤, 硫化氢, 自噬, LC3, 细胞凋亡, 印迹法, 蛋白质

Abstract: Objective To investigate the effects of hydrogen sulfide on autophagy and the apoptosis after acute spinal cord injury in rats. Methods Thirty-six adult male SD rats (250-300 g) were randomly divided into three groups (n=12 for each group): sham operation group (Sham group), spinal cord injury group (Model group) and hydrogen sulfide pre-treatment group (H2S group). Allens method was used to establish the rat model of spinal cord injury. Rats of sham operation group received only laminectomy. Rats of H2S group received sodium hydrosulphide injection intraperitoneally (50 μmol/kg) 1h after spinal cord injury, and Model group was given the same amount of saline solution. Rats in the three groups were sacrificed 24 h after spinal cord injury, then the spinal cord was removed. The expressions of LC3, p70S6K and Cleaved caspase-3 were detected by Western blot assay. The expression of LC3 was also detected by immunofluorescence. The cell apoptosis was assessed by TUNEL stain. Results Compared with Sham group, the expression levels of LC3Ⅱ/LC3Ⅰand Cleaved caspase3 were increased in Model group, but the expression of p70S6K decreased and cell apoptosis increased in Model group (P <0.01). Compared with Model group, the expression levels of LC3Ⅱ/LC3Ⅰand Cleaved caspase-3 were decreased significantly, while the expression of p70S6K increased and cell apoptosis decreased significantly in H2S group (P < 0.01). Conclusion Hydrogen sulfide can inhibit autophagy and reduce cell apoptosis after acute spinal cord injury in rats.

Key words: acute spinal cord injury, H2S, autophagy, LC3, apoptosis, blotting, western