尼莫地平联合依达拉奉对蛛网膜下腔出血大鼠海马神经细胞的保护作用

Neuroprotective effects of the combination nimodipine and edaravone on hippocampal neurons after subarachnoid hemorrhage in rats

目的探讨联合应用尼莫地平和依达拉奉对蛛网膜下腔出血大鼠海马神经细胞的保护作用。方法取健康SD雄性大鼠24只,按随机数字表法随机将大鼠分为模型组、假手术组、尼莫地平组及联合用药组(尼莫地平+依达拉奉),每组6只大鼠。线刺法制备SAH模型成功后,尼莫地平组立刻腹腔注射尼莫地平1mg/kg,联合用药组注射尼莫地平1mg/kg、依达拉奉3mg/kg;模型组与假手术组腹腔注射等量等渗盐水(6.5ml/kg)。术后24h按同样方法、剂量再给药1次。采用免疫组化和Western印记法检测大鼠SAH后48h,海马区Caspase-3、Bcl-2阳性细胞的表达情况。结果①免疫组化检测Caspase-3阳性细胞表达:假手术组、模型组、尼莫地平组及联合用药组分别为(2.2±1.7)%、(17.8±3.5)%、(12.7±3.5)%及(7.3±2.2)%;Bcl-2阳性细胞表达分别为(4.8±1.3)%、(15.7±4.5)%、(24.2±5.0)%及(39.8±5.6)%。各组比较差异均有统计学意义(P<0.05,P<0.01)。②Western印记检测结果(与β-actin密度比值):Caspase-3阳性细胞表达假手术组、模型组、尼莫地平组及联合用药组分别为0.09±0.07、0.65±0.10、0.46±0.12、0.28±0.12;Bcl-2阳性细胞表达分别为0.56±0.15、0.89±0.12、1.22±0.14、1.57±0.18,各组比较差异均有统计学意义(P<0.05,P<0.01)。结论尼莫地平或尼莫地平联合依达拉奉均可减少SAH后海马区Caspase-3阳性细胞表达,增加Bcl-2阳性细胞表达。尼莫地平联合依达拉奉的效果比单独应用尼莫地平的效果更佳。

Objective To observe the neuroprotective effects of the combination nimodipine and edaravone on hippocampal neurons after subarachnoid hemorrhage （SAH） in rats and their mechanisms. Methods Twenty-four male SD rats were randomly allocated into model, sham-operation, nimodipine, and combination edaravone and nimodipine groups according to a random number table （ 6 rats in each group）. Nimodipine 1 mg/kg was injected intrapefitoneally immediately after a model of SAH was induced successfully by the suture method in the nimodipine group; nimodipine 1 mg/kg and edaravone 3 mg/kg were injected intraperitoneally in the combination group; and the same amount of isotonic saline （ 6.5 ml/ kg） were injected intraperitoneally in the model and sham-operation groups. The same dose of medications were given again in the same way at 24 hours after the first injection. The expressions of caspase-3 and Bcl- 2 positive cells in rat hippocampal region were detected by immunohistochemical method and Western blot method. Results ①Immunohistochemical method：the expressions of caspase-3 in the sham-operation, model, nimodipine, and combination groups were 2. 2± 1.7, 17.8 ± 3.5, 12. 7 ± 3.5, and 7.3 ± 2. 2, respectively; the expressions of Bcl-2 positive cells were 4. 8 ± 1.3, 15, 7 ±4. 5, 24. 2 ±5.0, and 39. 8± 5.6, respectively. There were significant differences between all the groups（ P 〈 0. 05, P 〈 0. 01 ）. ②The results of Western blot method（ and β-actin density ratios） ： The expressions of caspase-3 positive cells in the sham-operation, model, nimodipine, and combination groups were 0. 09 ± 0. 07, 0. 65 ±0. 10, 0. 46±0. 12, and 0. 28 ± 0. 12, respectively; and the expressions of Bcl-2 positive cells were 0. 56 ±0. 15, 0. 89 ±0. 12, 1.22 ±0. 14, and 1.57 ±0. 18, respectively. There were significant differences between all the groups （P 〈 0. 05, P 〈 0. 01 ）. Conclusions Nimodipine alone or the combination edaravone and nimodipine may decrease the expressions of caspase-3 positive cells in hippocampal region after SAH, and increase the expressions of Bcl-2 positive cells. The efficacy of the combination edaravone and nimodipine is better than that of nimodipine alone.