电针对血管性认知障碍大鼠海马功能活动及其分子表达谱的影响

Effect of Electroacupuncture on Hippocampal Functional Activity and Molecular Expression Profile in Rats with Vascular Cognitive Impairment

目的:探讨“通督调神”电针对血管性认知障碍(VCI)模型大鼠海马功能活动及其海马全基因组表达谱的影响。方法:将24只健康雄性SD大鼠采用随机数字表法分为假手术组、模型组和电针组,每组8只。模型组、电针组采用双侧颈总动脉结扎法制备血管性认知障碍大鼠模型,假手术组仅分离双侧颈总动脉但不结扎。电针组大鼠取百会、神庭穴进行干预,电针频率1/20 Hz,30 min/次,1次/d,疏密波,共干预28 d,其他2组大鼠采取同等条件抓取,但不进行任何干预。采用Barnes迷宫测试评价大鼠空间学习记忆能力;采用Y迷宫测试评价大鼠空间工作记忆能力;采用小动物7.0 T磁共振静息态脑功能成像分析海马功能活动局部一致性(ReHo)变化;采用AgilentmRNA表达谱芯片分析海马全基因组差异表达情况。结果:①行为学分析结果:与假手术组比较,模型组Barnes迷宫逃避潜伏期明显上升(P<0.05),目标象限持续时间百分比明显下降(P<0.05),Y迷宫交替率明显下降(P<0.05)。与模型组比较,电针组Barnes迷宫逃避潜伏期明显下降(P<0.05),目标象限持续时间百分比明显上升(P<0.05),Y迷宫交替率明显增加(P<0.05)。②ReHo变化结果:与假手术组比较,模型组双侧前额叶、海马等脑区功能活动局部一致性明显下降(P<0.005);与模型组比较,电针组双侧海马、前额叶及梨状皮层等脑区功能活动局部一致性明显增强(P<0.005)。③基因组学结果:模型组与假手术组海马mRNA差异表达>2倍的基因共705个(P<0.05),其中195个基因(Sytl1、Trpv6、Klhl14、Npsr1、Myh3、Galnt3、Nr4a1、Bmp3、Egr2等)表达下调,510个基因(Insl6、Efcab1、Akr1b1、Tagln、Glrb、Akr1c13、Kcne1L、Ubap1等)表达上调。电针组与模型组海马mRNA差异表达>2倍的基因共399个(P<0.05),其中166个基因(Klhl14、Bmp3、Npsr1、Cacna1e等)表达上调,233个基因(Insl6、Ube2k、Cd⁃kn1c、Camp、Upk1b等)表达下调。结论:“通督调神”电针百会、神庭穴可以改善血管性认知障碍大鼠认知功能,其机制可能与增强海马、前额叶等脑区功能以及调控钙离子、能量代谢相关分子的表达参与调节神经活动等有关。

Objective: To explore the effect of "Tongdu Tiaoshen" electroacupuncture method on the hippocampal function and the gene expression profile of the hippocampal tissue in rats with vascular cognitive impairment. Methods: A total of 24 healthy male SD rats were randomly divided into the sham operation group, the model group and the electroacupuncture group, with 8 rats in each group. Bilateral common carotid artery ligation was used to prepare a rat model of vascular cognitive impairment in the model group and the electroacupuncture group, only bilateral common carotid artery was separated without ligation in the sham operation group. The electroacupuncture group was intervented by stimulating at Baihui(GV20) and Shenting(GV24) acupoints, the frequency of electroacupuncture was 1/20 Hz, 30 min each time, once a day, dilatational wave, continous intervention for 28 days, and the other two groups were grabbed under the same conditions without any other intervention. Barnes maze test was used to evaluate the spatial learning and memory ability of rats;Y maze test was used to evaluate the spatial working memory ability of rats;small animal 7.0 T magnetic resonance resting brain functional imaging was used to analyze the changes of regional homogeneity(ReHo) of hippocampal functional activity;Agilent mRNA expression microarray was used to analyze the differential gene expression of the whole hippocampus genome. Results:(1) Behavioral analysis results: compared with the sham operation group, the Barnes maze escape latency of the model group increased significantly(P<0.05), the target quadrant duration percentage decreased significantly(P<0.05), and the Y maze alternation rate decreased significantly(P<0.05);compared with the model group, the Barnes maze escape latency of the electroacupuncture group decreased significantly(P<0.05), the target quadrant duration percentage and the Y maze alternation rate increased significantly(P<0.05).(2) ReHo changes results: compared with the sham operation group, the ReHo of functional activitives of the bilateral prefrontal lobes, hippocampus and other brain regions in the model group decreased significantly(P<0.005);compared with the model group, the ReHo of functional activities of the bilateral hippocampus, prefrontal lobes and piriform cortex in the model group increased significantly(P<0.005).(3) Genomics results: a total of 705 genes with 2-fold differential expression of hippocampal mRNA between the model group and the sham operation group(P<0.05), of which 195 genes(such as Sytl1, Trpv6, Klhl14, Npsr1, Myh3, Galnt3, Nr4 a1, Bmp3, Egr2, etc.) were down-regulated, of which 510 genes(such as Insl6, Efcab1, Akr1 b1, Tagln, Glrb, Akr1 c13, Kcne1 L, Ubap1, etc.) were up-regulated. A total of 399 genes with 2-fold differential expression of hippocampal mRNA between the electroacupuncture group and the model group(P<0.05), of which 166 genes(such as Klhl14, Bmp3, Npsr1, Cacna1 e, etc.) were up-regulated, of which 233 genes(such as Insl6, Ube2 k, Cdkn1 c, Camp, Upk1 b,etc.) were down-regulated. Conclusion: The "Tongdu Tiaoshen" electroacupuncture at Baihui and Shenting acupoints can improve the cognitive function of rats with vascular cognitive impairment, and its mechanism may be related to enhance the function of hippocampus,prefrontal lobes and other brain regions, regulate the expression of calcium ions and energy metabolism-related molecules by regulating nerve activity activity.