归芪聪志汤对血管性痴呆模型大鼠行为学和海马组织基因表达谱的影响

Effects of Gui Qi Congzhi Decoction on Gene Expression Profile of Hippocampus in Rats with Vascular Dementia Model

目的研究归芪聪志汤对血管性痴呆(Vasculardementia,VD)大鼠模型海马基因表达谱的影响及部分筛选基因表达的影响。方法从120只SPF级Wistar雌性大鼠中随机选取10只作为假手术组,其余均采用改良后的两血管阻断法改良法制备血管性痴呆大鼠模型,经水迷宫筛选出合格造模大鼠35只,随机区组分为假手术组,模型组,阳性对照组,归芪聪志汤高、中、低剂量组。归芪聪志汤连续灌胃4周后处死大鼠取材海马组织。Morris水迷宫检测造模后及治疗后行为学变化。全基因组表达谱芯片检测归芪聪志汤中剂量组与模型组海马组织之间差异表达的基因;RT-qPCR检测紧密连接蛋白-1,2,3,14,22(claudin-1,claudin-2,claudin-3,claudin-14,claudin-22),细胞黏附分子(ICAM),血管紧张素转化酶(ACE)的mRNA表达量。结果Morris水迷宫检测显示:模型鼠逃避潜伏期明显延长,跨原平台次数明显减少(P<0.01)。治疗28d后模型组逃避潜伏期明显延长,跨原平台次数明显减少(P<0.05);与模型组相比,各治疗组大鼠逃逸潜伏期均缩短,跨原平台次数均增多(P<0.01),其中归芪聪志汤中剂量组、阳性药物对照组大鼠学习记忆能力改善情况优于归芪聪志汤高、低剂量组(P<0.05)。基因芯片和RT-qPCR检测结果显示:与假手术组比较,模型组claudin-1,claudin-2,claudin-3,claudin-22,ICAM,ACE基因表达量均升高(P<0.05,P<0.01),claudin-14、CDH3基因表达量均下降(P<0.05,P<0.01);与模型组比较,各治疗组claudin-1,claudin-2,claudin-3,claudin-22,ICAM,ACE基因表达量均下降,claudin-14,CDH3基因表达量均升高,其中,归芪聪志汤中剂量组、阳性药物对照组优于归芪聪志汤高、低剂量组(P<0.05)。结论归芪聪志汤能下调VD模型大鼠海马组织claudin-1,claudin-2,claudin-3,claudin-22,ICAM,ACE的表达,并升高claudin-14,CDH3的表达,从而发挥抗VD作用。

Objective To study gene expression profile of hippocampus in rats with vascular dementia（VD） model treated by Gui Qi Congzhi decoction. Methods A total of 120 SPF female Wistar rats were enrolled as experimental animals. Except 10 rats were selected as Sham group,all other rats were employed to establish VD model of rat with the method of‘ modified two vessel occlusion＇, and subsequently, water maze test was utilized to screen qualified model rats. Finally,a total of 35 qualified model rats were randomly divided into model group,positive control group,Gui Qi Congzhi decoction groups with high, middle, and low dosages. Morris water maze was used to detect the behavioral changes after completing model and treatment. After intragastric administration with Gui Qi Congzhi decoction for 4 weeks,rats were sacrificed to obtain hippocampus tissue. A rat genome microarray profile was chosen to screen differentially expressed genes between Gui Qi Congzhi decoction middle dosage group and model group;m RNA expression of all screened genes in hippocampus were then detected by real time quantitative PCR（RT-q PCR） to validate their expression trends screened by microarray. Results After completing model,compared with Sham group,the average escape latency of VD model group was significantly prolonged, and crossing platform number decreased significantly（P〈0.01）. After 4 courses of treatment, compared with the model group, rats escape latency in all treatment groups was shortened, crossing platform number were increased（P〈0.01）; of which Gui Qi Congzhi decoction medium dose group and positive control group were superior to Gui Qi Congzhi decoction low and high dose groups（P〈0.05）. Microarray and RT-q PCR analysis showed that：compared with the Sham group,the expression of claudin-1, claudin-2, claudin-3, claudin-22, ICAM and ACE in model group increased（P〈0.01）, and the expression of claudin-14,CDH3 decreased（P〈0.05,P〈0.01）. Compared with the model group,the expression of claudin-1, claudin-2, claudin-3, claudin-22, ICAM and ACE in the treatment groups decreased, and the expression of claudin-14,CDH3 increased. The treating effects of Gui Qi Congzhi decoction middle dosage group and positive drug group is better than other treating groups. Conclusion Gui Qi Congzhi decoction can significantly upregulate the expression of claudin-14 and CDH3 of hippocampus in VD model rats,and downregulate the expression of claudin-1,claudin-2,claudin-3,claudin-22,ICAM and ACE to exert its anti-vascular dementia action.