论文部分内容阅读
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 the effect of GuiQiChiZhiTang on the gene expression profile of hippocampus in vascular dementia (VD) rat model and the effect of partial screening of gene expression. Methods Totally 10 120 Wistar female SPF rats were randomly selected as the sham-operated group. The rest rats were all treated with the modified two-vessel blockade method to prepare the vascular dementia rat model. 35 rats were randomly divided into sham operation group, model group, positive control group, GuiQiChi Zhi Tang high, medium and low dose groups. Guiqi clever soup after continuous gavage 4 weeks after the rats were sacrificed hippocampal tissue. Morris water maze test after modeling and behavioral changes after treatment. Whole-genome cDNA microarray was used to detect the gene expression difference between GuijuTengzhi decoction middle dose group and model group hippocampus. RT-qPCR was used to detect the expression of claudin-1 (claudin- 2, claudin-3, claudin-14 and claudin-22), the expression of ICAM and ACE. Results Morris water maze test showed that the escape latency of model rats was significantly prolonged and the number of transplantaic platforms decreased significantly (P <0.01). Compared with the model group, the escape latency of rats in each treatment group was shortened and the number of transplants across platforms increased (P <0.01), and the escape latency was significantly prolonged in the model group after 28 days of treatment. The improvement of learning and memory abilities of Guqi Cong Zhi decoction medium dose group and positive drug control group were better than that of Gui Chi Cong Zhi Tang high and low dose group (P <0.05). The results of gene chip and RT-qPCR showed that the expression of claudin-1, claudin-2, claudin-3, claudin-22, ICAM and ACE in model group were significantly increased compared with sham operation group 0.01). The expression of claudin-1, claudin-2, claudin-3, claudin-22, ICAM, ACE-1 in all treatment groups were significantly lower than those in model group (P <0.05, P <0.01) The expression of claudin-14 and CDH3 genes were all increased, among which the Guigongzhizhi Decoction dose group and the positive drug control group were better than the Guqi Congzhi Decoction high and low dose groups (P <0.05) . Conclusions Guqi Cong Zhi decoction can down-regulate the expressions of claudin-1, claudin-2, claudin-3, claudin-22, ICAM and ACE in hippocampus of VD rats and increase the expressions of claudin-14 and CDH3, VD role.