侧脑室注射DIDS对缺血再灌注脑损伤大鼠神经元凋亡的拮抗作用*
Effect of intracerebroventricular injection with DIDS on neuronal apoptosis of rats with focal cerebral ischemia？？reperfusion injury

目的：观察侧脑室注射氯通道阻断剂DIDS对在体神经元凋亡的影响。方法：72只雄性SD大鼠随机分为假手术组、缺血再灌注（I/R）组、生理盐水组和DIDS组。除假手术组，余3组用线栓法建立大鼠局灶性缺血再灌注脑损伤模型。模型制备成功后生理盐水组和DIDS组分别立即通过侧脑室注射生理盐水和有效剂量的DIDS。72 h后TTC染色观察脑梗死灶面积，用HE染色观察脑组织形态学的变化，Western blot法观察海马CA1区活性Caspase？？3的表达。结果：假手术组无脑梗死灶出现，海马CA1区神经元排列较好，胞体饱满，折光性较好;余3组脑梗死灶面积差异有统计学意义（F=4.778，P=0.025）；I/R组和生理盐水组有明显的梗死灶出现，海马CA1区神经元排列错乱，水肿，间隙增宽;与I/R组和生理盐水组相比，DIDS组脑梗死面积缩小(P<0.05)，组织形态明显改善。4组海马CA1区活性Caspase？？3表达差异有统计学意义（F=166.700,P<0.001);假手术组有少量活性Caspase？？3表达；I/R组和生理盐水组活性Caspase？？3表达下降（P<0.05）;与I/R组和生理盐水组相比，DIDS组活性Caspase？？3表达降低（P<0.05）。结论：DIDS对在体神经元的凋亡有拮抗作用，对大鼠缺血再灌注脑损伤有保护作用。
Aim: To observe the effect of DIDS, a kind of chloride channel blocker, being injected from lateral ventricle, on neuronal apoptosis of rats with focal cerebral ischemia？？reperfusion(I/R) injury. Methods: A total of 72 rats were randomly divided into sham operation group, I/R group, saline group and DIDS group. The rats in I/R group, saline group and DIDS group were established focal cerebral I/R injury model by blocking the middle cerebral artery. The rats in saline group and DIDS group were given saline or the effective dose of DIDS by intracerebroventricular injection. After 72 h, TTC staining method was used to observe the brain infarct size, HE staining was used to observe the change of tissue morphology and the Western blot was used to observe the expression of activated Caspase？？3 in hippocampus CA1 area. Results: There were significant differences in infarct area among I/R group, saline group and DIDS group(F=4.778，P=0.025) and in activated Caspase？？3 expression among sham operation group, I/R group, saline group and DIDS group（F=166.700,P<0.001).No brain infarcts occurred and the neuron arrangement in hippocampus CA1 area was normal and little activated Caspase？？3 appeared in sham operation group. There were obvious infarcts,edema, broadening gap, and increasing activated Caspase？？3 expression and the neurons in hippocampus CA1 area arranged disorderly in I/R group and saline group(P<0.05). Compared with those of saline group and I/R group, the infarct size decreased, the morphology was improved, and the activated Caspase？？3 expression decreased in DIDS group(P<0.05). Conclusion: DIDS has protective effect on cerebral I/R injury and the antiapoptotic effect on neuronal apoptosis of rats in vivo