穴位埋线对血管性痴呆大鼠海马CA1区神经细胞凋亡的影响

Effects of acupoint catgut embedding therapy on nerve cell apoptosis in hippocampus CA1 area of rats with vascular dementia

目的观察穴位埋线疗法对血管性痴呆(VD)大鼠海马CA1区神经细胞凋亡和Bcl-2 mRNA及Bax mRNA表达的影响,探讨穴位埋线对VD大鼠脑缺血性损伤的神经保护机制。方法采用改良Pulsinelli’s 4血管阻断法建立VD大鼠模型,随机数表法分为VD模型组、穴位埋线组、尼莫地平组,并设置假手术组作为对照。两个治疗组分别施行穴位埋线和尼莫地平治疗。Morris水迷宫检测各组大鼠学习记忆能力后,取含海马的脑组织,TUNEL法检测海马CA1区神经细胞凋亡,原位杂交法检测其Bcl-2 mRNA及Bax mRNA表达,观察并比较各组大鼠海马神经细胞凋亡、蛋白表达的变化。结果 VD模型组大鼠海马CA1区可见大量凋亡细胞、Bcl-2 mRNA表达降低、Bax mRNA的表达增高,与假手术组比较差异均有统计学意义(P<0.01)。VD模型组表现出明显的学习记忆障碍,与假手术组相比差异有统计学意义(P<0.01);与VD模型组比较,穴位埋线组大鼠学习记忆能力显著提高(P<0.01),CA1区凋亡细胞明显减少(P<0.01),可见一定数量的Bcl-2 mRNA阳性细胞表达,而Bax mRNA阳性细胞表达较少。结论穴位埋线可通过上调VD大鼠海马CA1区Bcl-2 mRNA的表达、下调Bax mRNA的表达,抑制海马神经细胞凋亡,改善VD大鼠学习记忆能力。

Objective To observe the effect of Acupoint Catgut Embedding therapy（ACET） on nerve cell apoptosis and expression of Bel-2 and Bax protein in hippoeampus CA1 area of rats with vascular dementia and to explore their roles in the neuroprotection mechanism of cerebral ischemie injury. Methods VD models were established with the modified Pulsinelli four vessel occlusion （4-VO） method. The VD model rats were randomly divided into four groups by the stochastic indicator method： VD model group （group M） , ACET group （group A） and nimodipine group （group N） , and sham-operated group （ group S）. Two treatment groups were treated with ACET and nimodipine treatment. Learning and memory abilities of rats in 4 groups were detected with Morris Water Maze test after all rats ended treatment. The brain tissue containing hippocampus was collected, and the nerve cell apoptosis of CA1 area of hippocampus of rats was detected with TUNEL, and the expression of Bcl-2 and Bax protein in hippocampus CA1 area were detected with hybridization in situ. Changes of nerve cell apoptosis and content of Bcl-2 and Bax in hippocampus CA1 area were observed and compared. Results A large amount of apoptotic ceils showed. The content of Bcl-2 mRNA reduced, the content of Bax mRNA increased in hippocampal CA1 region of rats in group M, and the differences were statistically significant by comparing with group S（P 〈0.01 ）. The obvious learning and memory disorder appeared in group M, and the difference was significant by comparing with group S（P 〈0.01 ）. Compared with group M, the ability of learning and memorizing of rats in group A significantly increased（P 〈 0.01 ） and the apoptosis cells were significantly reduced（ P 〈 0.01 ）. Bcl-2 mRNA expression in the CA1 area of hippocampus of group A had a significant increasing but Bax mRNA expression decreased. Conclusion ACET may improve the ability of learning and memorizing regulation of Bcl-2 mRNA, down-regulation of Bax mRNA apoptosis of rats with vascular dementia. ability of VD rats, and its mechanism may be related to up- in hippocampus CA1 area and inhibition of the nerve cell