失荅剌知丸对脑缺血再灌注大鼠海马区神经干细胞增殖的影响

Effect of Shida Lazhi pill on the proliferation of neural stem cells in hippocampus of rats with cerebral ischemia reperfusion

目的观察失荅剌知丸对脑缺血再灌注大鼠海马区神经干细胞增殖情况的影响,探究其可能的作用机制。方法将204只SD雄性大鼠随机分为空白组(12只)、假手术组(12只)、模型组(36只)、尼莫地平组(36只)、失荅剌知丸低剂量组(36只)、失荅剌知丸中剂量组(36只)及失荅剌知丸高剂量组(36只),空白组不进行手术操作,假手术组进行假手术操作,其余各组均采用改良Longa法建立大鼠脑缺血再灌注模型。造模成功后各组大鼠均按1 mL/100 g剂量给药,尼莫地平组予浓度为1.08 mg/mL的尼莫地平混悬液灌胃,失荅剌知丸低、中、高剂量组分别给予浓度为1.5、3.0、4.5 g/mL的失荅剌知丸药液灌胃,空白组、假手术组及模型组予0.9%氯化钠注射液灌胃。分别观察各组大鼠在灌胃1、3、7 d后神经功能Garcia JH评分、海马区神经干细胞增殖情况及脑组织巢蛋白(Nestin)表达情况的变化。结果与空白组及假手术组同期比较,模型组、尼莫地平组及失荅剌知丸低、中、高剂量组Garcia JH评分均明显降低(P<0.05),神经干细胞增殖增多(P<0.05),Nestin蛋白表达增高(P<0.05),并且随着时间的延长效果越明显;与模型组同期比较,尼莫地平组及失荅剌知丸低、中、高剂量组Garcia JH评分均明显升高(P<0.05),神经干细胞增殖增多(P<0.05),Nestin蛋白表达增高(P<0.05);与尼莫地平组同期比较,失荅剌知丸低剂量组Garcia JH评分升高、神经干细胞增殖增多及Nestin蛋白表达增高情况均不及尼莫地平组(P<0.05),失荅剌知丸中剂量组与尼莫地平组疗效相当(P>0.05),失荅剌知丸高剂量组疗效则明显优于尼莫地平组(P<0.05)。结论失荅剌知丸对脑缺血再灌注大鼠的神经功能损伤具有确切疗效,可明显改善大鼠神经功能损伤症状,促进Nestin蛋白表达,其作用机制可能与促进神经干细胞增殖有关,并且其促进作用具有剂量和时间依赖性。

Objective To observe the effects of Shida Lazhi pill on the proliferation of neural stem cells in hippocampus of rats with cerebral ischemia reperfusion,and explore the possible mechanism. Methods 204 male SD rats were randomly divided into blank group( 12),sham-operation group( 12),model group( 36),nimodipine group( 36),low-dose Shida Lazhi pill group( 36),medium-dose Shida Lazhi pill group( 36) and high-dose Shida Lazhi pill group( 36). The blank group did not operate,the sham operation group did the sham operation,the other groups were all treated by modified Longa method to establish cerebral ischemia reperfusion model in rats. After the success of modeling,all rats were given 1 mL/100 g doses. Rats in nimodipine group were given the nimodipine suspension with a concentration of 1. 08 mg/mL by gastrogavage. The low-dose Shida Lazhi pill group,medium-dose Shida Lazhi pill group,high-dose Shida Lazhi pill group were respectively given liquid Shida Lazhi pill with concentration of 1. 5 g/mL,3 g/mL and 4. 5 g/mL by gastrogavage. The blank group,sham-operation group and model group were given 0. 9% sodium chloride injection by gastrogavage. The Garcia JH score,the proliferation ofhippocampal neural stem cells and the expression of nestin in brain tissue were observed after 1,3,and 7 days after gastrogavage. Results Compared with the blank group and the sham-operation group,the Garcia JH scores of the model group,the nimodipine group,and the low-dose,medium-dose,high-dose groups of Shida Lazhi pill were significantly decreased( P < 0. 05),and the proliferation of neural stem cells were increased( P < 0. 05),Nestin protein expression increased( P < 0. 05),and the effect was more obvious with the prolongation of time. Compared with the model group,the Garcia JH scores in the nimodipine group and the low-dose,medium-dose,high-dose groups of Shida Lazhi pill were significantly increased( P < 0. 05),the proliferation of neural stem cells increased( P< 0. 05) and the expression of Nestin protein increased( P < 0. 05). Compared with the nimodipine group,the increase of Garcia JH score,neural stem cell proliferation and Nestin protein in low-dose Shida Lazhi pill group were not as good as the nimodipine group( P < 0. 05),and the effect of medium-dose Shida Lazhi pill group was equivalent to that of the nimodipine group( P < 0. 05),and the effect of the high-dose Shida Lazhi pill group was obviously superior to that of the nimodipine group( P < 0. 05). Conclusion Shida Lazhi pill has a definite curative effect on neurological impairment in rats with cerebral ischemia-reperfusion,can significantly improve the symptoms of neurological impairment in rats,promote the expression of Nestin protein,and its mechanism may be related to the promotion of neural stem cell proliferation,and the effect has a dose and time dependent effect.