不同时间窗高压氧干预对大鼠脊髓损伤后细胞凋亡及脊髓功能恢复的影响

doi:10.13481/j.1671-587x.20150328

摘要/Abstract

摘要：

目的:观察不同时间窗高压氧干预下的大鼠脊髓损伤(SCI)后细胞凋亡和脊髓功能的恢复情况,探讨最佳的高压氧治疗方法(HBOT),为临床治疗SCI提供实验依据。方法:160只SD大鼠随机分为模型对照组、HBO-PC组、HBOT组和HBO-PC+HBOT组。采用Allen's打击法建立SCI模型。模型对照组大鼠仅制模,未进行任何干预;HBO-PC组大鼠于高压氧干预10 d后立即制模,制模后未进行任何干预;HBOT组大鼠于制模后立即行高压氧干预;HBO-PC+HOBT组大鼠于高压氧干预10 d后立即制模,制模后继续高压氧干预。各组大鼠分别于制模后第1、2、7和14天取材,免疫组织化学PV-9000二步法检测大鼠Fas和FasL的表达水平,光镜下观察,并对结果进行统计学分析;采用TUNEL法检测细胞凋亡指数(AI),神经功能评价采用BBB评分。结果:与模型对照组比较,HBO-PC组、HBOT组和HBO-PC+HBOT组大鼠各时间点Fas和FasL表达水平均降低,且差异均有统计学意义(P<0.01);HBO-PC+HBOT组大鼠各时间点Fas和FasL表达水平比HBOT组和HBO-PC组降低,且差异有统计学意义(P<0.05);HBOT组大鼠各时间点Fas和FasL表达水平与HBO-PC组比较差异无统计学意义(P>0.05)。与模型对照组比较,其他各组大鼠细胞AI明显降低,差异均有统计学意义(P<0.01);HBO-PC+HBOT组AI低于HBOT组和HBO-PC组,且差异有统计学意义(P<0.05);HBOT组AI与HBO-PC组比较差异无统计学意义(P>0.05)。术后7和14 d HBO-PC组、HBOT组和HBO-PC+HBOT组大鼠BBB评分均高于模型对照组,且差异有统计学意义(P<0.01);HBO-PC+HBOT组BBB评分高于HBOT组和HBO-PC组,且差异有统计学意义(P<0.05或P<0.01);HBOT组BBB评分高于HBO-PC组,且差异有统计学意义(P<0.05)。结论:高压氧干预可减少SCI后细胞凋亡,促进脊髓功能的恢复;其治疗时间窗应提前到损伤前,HBO-PC联合HBOT法是治疗SCI的较佳方案。

关键词: 脊髓损伤, 高压氧, 干预时间窗, Fas, FasL, 细胞凋亡

Abstract:

Objective To study the effects of hyperbaric oxygen at different intervention time on the apoptosis and functional recovery following spinal cord injury(SCI) in the rats and to investigate the ideal application method of hyperbaric oxygen, and to provide some experimental basis for clinical hyperbaric oxygen therapy for SCI. Methods 160 SD rats were randomly divided into normal injury group, hyperbaric oxygen preconditioning(HBO-PC) group, hyperbaric oxygen therapy(HBOT)group and HBO-PC+HBOT group.The rat models of SCI were established with Allen's dropping weight technique.The rats in normal injury group weren't treated with any intervention;the rats in HBO-PC group were treated with hyperbaric oxygen intervention 10 d before modeling;the rats in HBOT group were treated with hyperbaric oxygen intervention only after modeling;the rats HBO-PC+HBOT group were treated with hyperbaric oxygen intervention before and after modeling.The segments of injured spinal cord were collected from the rats in 4 groups on the 1st, 2nd, 7th and 14th day after injury.The expression levels of Fas and FasL were detected with immunnohistochemistry PV-9000 two steps method.The apoptotic index(AI) was detected with TUNEL technique.The neurological function of spinal cord was assessed by BBB score. Results Compared with normal injury group, the expression levels of Fas and FasL of the rats in HBO-PC group, HBOT group and HBO-PC+HBOTgroup were decreased significantly(P<0.01);there were significant differences between HBO-PC+HBOT group and HBO-PC group, HBOT group(P<0.05), and there was no significant difference between HBO-PC group and HBOT group(P>0.05).Compared with normal injury group, the AI of the cells in HBO-PC group, HBOT group and HBO-PC+HBOT group were decreased significantly(P<0.01);compared with HBO-PC group and HBOT group, the AI of the cells in HBO-PC+HBOT group was decreased significantly(P<0.05);there was no significant difference between HBO-PC group and HBOT group(P>0.05).Compared with normal injury group, the BBB scores of the rats in HBO-PC group, HBOT group and HBO-PC+HBOT group 7 and 14 d after operation were increased significantly(P<0.01);compared with HBO-PC group and HBOT group, the BBB scores in HBO-PC+HBOT group were increased significantly (P<0.05 or P<0.01);compared with HBO-PC group, the BBB scores in HBOT group were increased significantly (P<0.05). Conclusion Hyperbaric oxygen can inhibit the apoptosis after SCI, which contributes to the recovery of locomotor performances in the rats.The best intervention time is in front of the damage.The method of HBO-PC combined with HBOT is the best way for the treatment of SCI.

Key words: spinal cord injury, hyperbaric oxygen, intervention time, Fas, FasL, apoptotosis

中图分类号:

R651.2

刘芳, 陈佳, 苏华. 不同时间窗高压氧干预对大鼠脊髓损伤后细胞凋亡及脊髓功能恢复的影响[J]. 吉林大学学报(医学版), 2015, 41(03): 578-583.

LIU Fang, CHEN Jia, SU Hua. Effects of hyperbaric oxygen at different intervention time on apoptosis and functional recovery following spinal cord injury in rats[J]. Journal of Jilin University Medicine Edition, 2015, 41(03): 578-583.

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