栀子苷对大鼠脊髓缺血再灌注后脊髓水肿及神经元细胞凋亡的影响

Effects of geniposide on the edema and neuronal apoptosis in spinal cord ischemia/reperfusion injury

目的探索栀子苷(GEN)对大鼠脊髓缺血再灌注(I/R)后脊髓水肿及神经元细胞凋亡的影响。方法腹主动脉夹闭法制备大鼠脊髓缺血再灌注损伤模型,实验分3组:假手术组(只切开不制作I/R),模型组(等量生理盐水预处理)和实验组(100 mg/kg栀子苷预处理),分别于术后24 h和48 h,利用BBB评分标准评价各组大鼠下肢运动功能,干湿法检测脊髓组织水肿指数,比色法检测脊髓组织中Ca^(2+)浓度,Western blot法检测AQP4蛋白表达,TUNEL染色法检测脊髓神经元细胞凋亡数目。结果 BBB评分显示,术后24 h,实验组与模型组下肢运动功能评分无显著差异(P>0.05),而术后48 h BBB评分实验组显著高于模型组(P<0.05);24 h和48 h,脊髓组织水肿指数,实验组均显著低于模型组(P<0.05);Ca^(2+)浓度实验组均显著低于模型组(P<0.05);Western blot显示,24 h和48 h后AQP4蛋白表达,实验组均显著低于模型组(P<0.05);TUNEL染色显示,24 h和48 h后实验组阳性细胞数均显著低于模型组(P<0.05)。结论栀子苷可通过抑制Ca^(2+)及AQP4蛋白表达减轻大鼠脊髓缺血再灌注后脊髓水肿及神经元细胞凋亡。

Objective To explore the effects of geniposide on the edema and neuronal apoptosis in spinal cord ischemia/ reperfusion inju-ry. Methods Spinal cord ischemia/ reperfusion injury model was made by abdominal aortic clipping method. All rats were divided into 3 groups, the sham group, the model group and the test group. Lower limb motor function was valued at the 24 h and 48 h post surgery via BBB scores. Spi-nal cord edema was detected via dry and wet method. Ca2＋ concentration in spinal cord tissue was valued by colorimetric method. Meanwhile, the protein expression of AQP4 was detected via western blot. Neuronal apoptosis was valued by TUNEL staining. Results At the 24h , there was no significantly difference between the test group with the model group on BBB scores （ P 〉0. 05） , while at the 48 h , BBB scores in the test group was markedly higher than the model group （ P 〈0.05） . Spinal cord edema and Ca2 ＋ concentration in the test group were significantly lower than the model group both at 24 h and 48 h （ P 〈 0. 05） . Western blot showed that, the protein expression of AQP4 in the test group were significantly lower than the model group both at 24 h and 48 h （ P 〈0.05 ） . TUNEL staining showed that the number of apoptotic cells in the test group were significantly lower than the model group both at 24 h and 48 h （ P〈0. 05 ） . Conclusion Geniposide could markedly suppressed spinal cord ede-ma and neuronal apoptosis in spinal cord ischemia/ reperfusion injury.