摘要
目的 观察腺苷对大鼠脊髓缺血再灌注损伤后细胞凋亡及B细胞白血病/淋巴瘤-2（bcl-2）和B细胞白血病/淋巴瘤-2相关蛋白X（bax）表达的影响,探讨其对脊髓损伤的保护作用.方法 通过阻断大鼠肾动脉水平的腹主动脉90 min建立大鼠脊髓缺血再灌注损伤模型.27只成年健康雄性SD大鼠随机分成3组,每组9只.实验分组如下：A组：假手术组;B组：缺血再灌注损伤组;C组：腺苷预处理组.每组分别设置再灌注12、24、48h3个时间点,于各时间点观察大鼠并对其后肢运动功能进行评分,取再灌注48 h大鼠的L2～L5节段脊髓组织作苏木素-伊红（HE）染色及尼氏（Nissl）染色,透射电镜观察超微结构,Western blot、实时定量逆转录聚合酶链反应（RT-qPCR）分别检测bcl-2和bax蛋白及mRNA的表达.结果 HE染色及Nissl染色结果显示：C组细胞水肿、尼氏体溶解等组织形态学变化较B组轻;透射电镜观察结果显示：C组的神经元超微结构损伤较B组轻;Western blot及RT-qPCR检测结果显示：腺苷能降低bax的表达,增强bcl-2的表达.结论 腺苷可减轻脊髓缺血再灌注损伤所造成的损害,促进神经功能的恢复.
Objective To study the expression levels of B lymphocytes/leukemia-2 （bcl-2） and bcl-2 associated X protein （bax） following ischemia-reperfusion injury and their action mechanism on apoptosis for investigating the protection of spinal cord I-R injury by means of adenosine preconditioning.Methods The experimental model was established in rats through cross-clamping of abdominal aorta inferior to the left renal artery.Twenty-seven rats were randomly divided into three groups：group A （n =9）,sham group; group B （n =9）,I-R group ; and group C （n =9）,administration with adenosine group.Spinal cord function was assessed by the Modified Tarlov Scale at 12,24 and 48 h after reperfusion.At 48 h after reperfusion,histologic analysis was done by hematoxylin and eosin （HE） and Nissl staining,ultrastructures were observed under transmission electron microscopy,and the expression levels of bcl-2 and bax pro tein and mRNA were detected by using Western blotting and real-time reverse transcriptase-polymerase chain reaction （RT-qPCR） respectively.Results HE staining and Nissl staining results showed：group B showed extensive spinal cord injury such as cell edema and dissolution of Nissl substance when compared to group C; the number of apoptotic cells in group C was less than that in group B.Western blotting and RT-qPCR results showed adenosine could reduce bax expression and increase the bcl-2 expression.Conclusion Adenosine can reduce spinal cord I-R and improve neurological function.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第12期2664-2667,共4页
Chinese Journal of Experimental Surgery
基金
国家科技部国际合作项目（2008DFA31140、2010DFA32660）
国家科技支撑计划资助项目（2011BAI11B22）
广东省自然科学基金资助项目（10251008002000002、10151008002000003）
广东省科技计划资助项目（2009[627]-2）
