复智胶囊对血管性痴呆模型大鼠海马神经元细胞凋亡时效的影响
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摘要
目的:观察复智胶囊对血管性痴呆大鼠海马区自噬正相关蛋白Beclin-1表达的影响及在海马神经元细胞凋亡中的作用。方法:雄性SD大鼠采用双侧颈总动脉分次结扎法,建立血管性痴呆大鼠模型。选取造模成功的大鼠,随机分为模型组、复智胶囊高剂量组、复智胶囊低剂量组及多奈哌齐组,并设假手术组。分别在第3天、第7天、第14天采用Western Blot法检测大鼠海马区脑组织中Beclin-1、Bcl-2和Caspase-3的表达,采用TUNEL法检测海马神经元细胞凋亡数。结果:术后3 d即对海马神经元细胞造成损害,与假手术组比较,造模后各组各时点的Beclin-1、Bcl-2和Caspase-3蛋白表达水平明显增高(P<0.05),海马神经元细胞大量凋亡(P<0.05)。与模型组比较,相同时点复智胶囊高剂量组、低剂量组Beclin-1和Bcl-2蛋白表达水平明显增高(P<0.05),Caspase-3蛋白表达水平明显下降(P<0.05),海马神经元细胞凋亡数目明显减少(P<0.05)。结论:复智胶囊对血管性痴呆模型大鼠海马神经元有一定保护作用,可能是通过上调Beclin-1蛋白表达水平、激活细胞自噬及抑制细胞凋亡来实现的。
Objective: To observe the effect of Fuzhi Capsule on the expression of autophagy-related protein Beclin-1 in hippocampus of rats with vascular dementia and the role of Fuzhi Capsule in apoptosis of hippocampal neurons. Methods: Rat model of vascular dementia was established by ligating bilateral common carotid arteries in male SD rats. Rats with successful model were randomly divided into model group,Fuzhi Capsule high-dose group,Fuzhi Capsule low-dose group and donepezil group,and sham operation group. The expression of Beclin-1,Bcl-2 and Caspase-3 in rat hippocampus was detected by Western Blot method on the3 rd,7 th and 14 th day respectively. The apoptotic number of hippocampal neurons was detected by TUNEL method. Results: 3 days after operation,the hippocampal neurons were damaged. Compared with the sham-operated group,the expressions of Beclin-1,Bcl-2 and Caspase-3 protein were significantly increased at each time point( P < 0. 05),and a large number of hippocampal neurons were apoptotic. Compared with the model group,the expression of Beclin-1 and Bcl-2 protein increased significantly( P < 0. 05),the expression of Caspase-3 protein decreased significantly( P < 0. 05) and the number of hippocampal neuron apoptosis decreased significantly( P < 0. 05) in the high-dose group and the low-dose group at the same time point. Conclusion: Fuzhi Capsule has protective effect on hippocampal neurons of vascular dementia model rats,which may be achieved by up-regulating the expres-sion of Beclin-1 protein,activating cell autophagy and inhibiting cell apoptosis.
Objective: To observe the effect of Fuzhi Capsule on the expression of autophagy-related protein Beclin-1 in hippocampus of rats with vascular dementia and the role of Fuzhi Capsule in apoptosis of hippocampal neurons. Methods: Rat model of vascular dementia was established by ligating bilateral common carotid arteries in male SD rats. Rats with successful model were randomly divided into model group,Fuzhi Capsule high-dose group,Fuzhi Capsule low-dose group and donepezil group,and sham operation group. The expression of Beclin-1,Bcl-2 and Caspase-3 in rat hippocampus was detected by Western Blot method on the3 rd,7 th and 14 th day respectively. The apoptotic number of hippocampal neurons was detected by TUNEL method. Results: 3 days after operation,the hippocampal neurons were damaged. Compared with the sham-operated group,the expressions of Beclin-1,Bcl-2 and Caspase-3 protein were significantly increased at each time point( P < 0. 05),and a large number of hippocampal neurons were apoptotic. Compared with the model group,the expression of Beclin-1 and Bcl-2 protein increased significantly( P < 0. 05),the expression of Caspase-3 protein decreased significantly( P < 0. 05) and the number of hippocampal neuron apoptosis decreased significantly( P < 0. 05) in the high-dose group and the low-dose group at the same time point. Conclusion: Fuzhi Capsule has protective effect on hippocampal neurons of vascular dementia model rats,which may be achieved by up-regulating the expres-sion of Beclin-1 protein,activating cell autophagy and inhibiting cell apoptosis.
引文
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[2]WANG H,JIANG R,HE Q,et al.Expression pattern of peroxisome proliferator activated receptors in rat hippocampus following cerebral ischemia and reperfusion injury[J].PPAR Res,2012,29(11):2075-2085.
[3]KOURTIS N,TAVERNARAKIS N.Autophagy and cell death in model organisms[J].Cell Death Differ,2009,16(1):21-30.
[4]马明,赵晴,杜建时,等.大鼠颈总动脉分次结扎建立血管性痴呆模型评价[J].中国老年学杂志,2012,32(1):72-74.
[5]徐世军,赵宜军,张文生,等.从中医脑络功能演变谈轻度认知障碍的病机[J].中医杂志,2011,52(19):1627-1629.
[6]李俊仙,肖红玲,王霞,等.中药治疗血管性痴呆的实验研究进展[J].中西医结合心脑血管病杂志,2010,8(1):89-90.
[7]马云枝,史继鑫,孟闯,等.复智胶囊对血管性痴呆大鼠脑组织海马神经元细胞凋亡的影响[J].时珍国医国药,2011,22(9):2177-2178.
[8]DECUYPERE J P,PARYS J B,BULTYNCK G.Regulation of the autophagic Bcl-2/Beclin-1 interaction[J].Cells,2012,143(3):284-312.
[9]HAILEY D W,RAMBOLD A S,SATPUTE-KRISHNAN P,et al.Mitochondria supply membranes for autophagosome biogenesis during starvation[J].Cell,2010,141(7):656-667.
[10]MEIJER A J,CODOGNO P.Regulation and role of autophagy in mammalian cells[J].Int J Biochem Cell Biol,2004,36(12):2445-2462.
[11]CARLONI S,BUONOCORE G,BALDUINI W.Proteetive role of autophagy in neonatal hypoxia-ischemia induced brain injury[J].Neurobiol Dis,2008,32(2):329-339.
[12]GABRYEL B,KOST A,KASPROWSKA D.Neuronal autophagy in cerebral ischemia:a potential target for neuroprotective strategies[J].Pharmacol Rep,2012,64(1):1-15.
[13]KOIKE M,SHIBATA M,TADAKOSHI M,et al.Inhibition of autophagy prevents hippocampal pyramidal neuron death after hypoxic-ischemic injury[J].Am J Pathol,2008,172(2):454-469.
[14]PARK H K,CHU K,JUNG K H,et al.Autophagy is involved in the ischemic preconditioning[J].Neurosci Lett,2009,451:16-19.