南方医科大学学报 ›› 2014, Vol. 34 ›› Issue (01): 92-.

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腺苷预处理对大鼠脊髓缺血再灌注损伤的保护作用

富 旗,周正芳,李小辉,郭惠明,范小平,陈寄梅,庄 建,郑少忆,朱 平   

  • 出版日期:2014-01-20 发布日期:2014-01-20

Protective effect of adenosine preconditioning against spinal cord ischemia-reperfusion
injury in rats

  • Online:2014-01-20 Published:2014-01-20

摘要: 目的通过观察腺苷对大鼠脊髓缺血再灌注损伤后细胞凋亡及葡萄糖调节蛋白-78(GRP-78)和半胱氨酸天冬氨酸特异性
蛋白酶12(caspase 12)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:假手术组(A组);缺血再灌注损伤组(B组);腺苷预处
理组(C组)。于再灌注6、12、24 h观察大鼠并对其后肢运动功能进行评分,取再灌注24 h大鼠的L2-L5节段脊髓组织作HE染
色及TUNEL 染色,Western blot检测GRP-78和caspase 12蛋白的表达。结果HE染色结果显示:C组细胞水肿等组织形态学变
化较B组轻;TUNEL染色结果显示:与A组比较,B组的凋亡细胞数量显著增加;与B组相比,C组凋亡细胞数量较少。WB检
测结果显示:相较于B组,C组GRP-78蛋白表达增多(P<0.01),caspase 12蛋白表达减少(P<0.01)。结论腺苷通过上调GRP-78
的表达,下调caspase 12的表达,抑制细胞凋亡而发挥其保护作用。

Abstract: Objective To study the effect of adenosine preconditioning on cell apoptosis and expressions of glucose-regulated
protein (GRP-78) and cysteinylaspartate-specific protease 12 (caspase-12) in rats with spinal cord ischemia-reperfusion injury.
Methods Twenty-seven rats were randomized into 3 equal groups and subjected to sham operation (group A), spinal cord
ischemia-reperfusion injury (group B), or ischemia-reperfusion injury with adenosine treatment. Spinal cord
ischemia-reperfusion injury was induced by cross-clamping of the abdominal aorta inferior to the left renal artery. The spinal
cord function was assessed using the Modified Tarlov Scale at 6, 12, and 24 h after reperfusion. At 24 h after reperfusion,
histological analysis was carried out with HE staining; cell apoptosis and viability were determined with TUNEL staining, and
the expressions of GRP-78 and caspase-12 proteins were determined with Western blotting. Results HE staining of the spinal
cord showed extensive spinal cord injury such as cell edema in group B as compared with group C. Compared with group A,
group B showed a significantly increased number of apoptotic cells; the number of apoptotic cells in group B was greater than
that in group C. Compared with group B, group C showed significantly increased GRP-78 expression (P<0.01) and decreased
caspase-12 expression (P<0.01). Conclusion Adenosine can up-regulate GRP-78 expression and down-regulate caspase-12
expression, and protects the spinal cord against ischemia-reperfusion injury by inhibiting cell apoptosis.