当归芍药散通过PI3K/AKT信号通路抗血管性痴呆的作用机制

The Mechanism of Anti-vascular Dementia of Danggui Shaoyao Powder via PI3K/AKT Signaling Pathway

目的探讨当归芍药散(DSS)通过PI3K/AKT信号通路抗血管性痴呆(VD)的作用机制。方法 SD大鼠随机分为空白组、假手术组、模型组、阳性药组(尼莫地平组,9.45 mg·kg-1)及DSS高、中、低剂量组(6.4、3.2、1.6g·kg-1);采用双侧颈总动脉永久性结扎法建立VD大鼠模型;大鼠按上述剂量灌胃给药,给药体积10mL·kg-1,空白组、假手术组、模型组给予等量生理盐水,每天1次,连续灌胃1个月。采用Morris水迷宫实验检测VD大鼠的学习记忆能力;采用ELISA法检测海马组织中肿瘤坏死因子-α(TNF-α)、核因子-κB(NF-κB)含量;采用WesternBlot法检测海马组织PI3K、AKT、p-AKT及LC3蛋白的表达;采用qPCR法检测海马组织PI3K、AKTmRNA的表达水平。结果与假手术组比较,模型组第1～4天的逃避潜伏期均明显延长,穿越平台的次数明显减少(P <0.01);海马组织中的TNF-α、NF-κB含量均显著增加(P <0.01),PI3K、AKT、pAKT蛋白表达量均显著降低(P <0.05,P <0.01),LC3-II/LC3-I值明显增高(P <0.01),PI3K、AKT mRNA表达水平均明显降低(P <0.01)。与模型组比较,DSS高、中剂量组第1～4天的逃避潜伏期均明显缩短,穿越平台的次数明显增加(P <0.05,P <0.01);DSS高剂量组的TNF-α、NF-κB含量均显著降低(P <0.05,P <0.01);DSS高剂量组的PI3K蛋白表达量显著升高(P <0.05),DSS高、中剂量组的AKT蛋白表达量显著升高(P <0.05),DSS高、中、低剂量组的p-AKT蛋白表达量均显著升高(P <0.05,P <0.01),DSS高、低剂量组的LC3-II/LC3-I值明显降低(P <0.01);DSS高、中剂量组的PI3K mRNA表达水平显著提高(P <0.05,P <0.01),DSS中剂量组的AKT mRNA表达水平显著提高(P <0.01)。结论 DSS能改善VD大鼠的学习记忆能力,可能通过PI3K/AKT信号通路发挥对VD的治疗作用。

Objective To study the mechanism of the anti-vascular dementia(VD) by Danggui Shaoyao powder(DSS) via PI3 K/AKT signaling pathway. Methods SD rats were randomly divided into blank group, Sham operation group,model group,positive drug group(Nimodipine group,9.45 mg·kg^-1) and DSS high,medium and low dose groups(6.4,3.2,1.6 g·kg^-1). The rat models of VD were established by permanent ligation of bilateral common carotid artery. Rats were given gastric administration at correspondent drugs and dosages,and the dosing volume is 10 mL·kg^-1;the blank group,Sham operation group and model group were given the same amount of saline once a day for one month. Morris water maze was used to detect the learning and memory abilities of VD rats;the content of tumor necrosis factor-alpha(TNF-α) and nuclear factor NF(NF-κB) in hippocampal tissue was detected by Elisa;Western Blot was used to detect the expression of PI3 K,AKT,p-AKT and LC3 proteins in hippocampus;and detection of PI3 K and AKT mRNA expression in hippocampus was carried out by qPCR. Results Compared with the Sham operation group,the escape latency of the model group was significantly prolonged on the1 st to 4 th day,and the number of times of crossing the platform was significantly reduced of TNF-α and NF-κB in hippocampus increased significantly(P < 0.01),the expression of PI3 K,AKT and pAKT decreased significantly(P < 0.05,P < 0.01),LC3-II/LC3-I increased significantly(P < 0.01),and the expression levels of PI3 K and AKT decreased significantly latency of DSS high and medium dose groups was significantly shortened in the 1 st to 4 th day,and the number of times crossing the platform was significantly increased(P < 0.05,P < 0.01);the contents of TNF-α and NF-κB in the high dose group of DSS were significantly decreased(P < 0.05,P < 0.01);the expression of PI3 K protein was significantly increased in DSS high dose group high and medium dose group(P < 0.05),and p-AKT protein was significantly increased in DSS high,medium and low dose group significantly decreased(P < 0.01),the PI3 K expression levels in the high and medium dose groups of DSS were significantly increased(P < 0.05,P < 0.01),and the AKT expression levels in the medium dose group of DSS were significantly increased which may play a therapeutic role in VD through PI3 K/AKT signaling pathway.