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目的：探讨天麻钩藤饮对高血压合并血管性认知障碍模型大鼠的干预作用，观察该方对磷脂酰肌醇3-激酶（PI3K）/丝氨酸/苏氨酸激酶（AKT）信号通路的影响，揭示其潜在的功能机制。方法：36只雄性SHR大鼠采用严格的随机数字表法进行分组，分为假手术组、模型组、尼莫地平组（9.375 mg/kg）和天麻钩藤饮低、中、高剂量组（5.938 g/kg、11.875 g/kg、23.750 g/kg），另取6只Wistar大鼠作为空白对照组，采用单侧颈总动脉阻断加高盐饮食方法建立高血压合并血管性认知障碍大鼠模型，随后应用尼莫地平和不同剂量的天麻钩藤饮连续治疗28 d。Morris水迷宫实验测试各组大鼠学习阶段的逃避潜伏期及测试阶段的目标象限停留时间、穿越平台次数，每周检测各组大鼠血压，尼氏（Nissl）染色观察大鼠海马CA1区神经元的病理改变及尼氏小体，原位末端标记法（TUNEL）染色用来评估对大鼠海马CA1区神经元凋亡的影响，蛋白质印迹法（Western Blotting）检测各组大鼠海马组织内AKT和PI3K的蛋白表达及磷酸化水平，酶联免疫吸附实验（ELISA）检测各组大鼠海马组织内Caspase-3、Bcl-2、Bax的蛋白表达及血清中ET-1的蛋白表达。结果：与空白组、假手术组相比，模型组大鼠血压持续升高、平均逃避潜伏期延长、目标象限停留时间和穿越平台次数减少（P<0.01）；海马Caspase-3、Bax蛋白表达明显上升（P<0.01）,Bcl-2蛋白表达下降（P<0.01）；大鼠海马细胞计数减少（P<0.01），尼氏小体减少，TUNEL阳性细胞增加（P<0.01）;AKT、PI3K蛋白磷酸化水平显著降低（P<0.01）。与模型组相比，天麻钩藤饮高、中剂量组和尼莫地平组大鼠收缩压和舒张压降低、平均逃避潜伏期减少、目标象限停留时间延长、穿越平台次数增加（P<0.01）；天麻钩藤饮高、中、低剂量组及尼莫地平组大鼠Caspase-3、Bax蛋白表达均下降（P<0.01）,Bcl-2蛋白表达上升（P<0.01）；大鼠海马细胞计数增加（P<0.01），尼氏小体增多，TUNEL阳性细胞降低（P<0.01）;AKT、PI3K蛋白磷酸化水平显著升高（P<0.01）。结论：天麻钩藤饮可能通过促进AKT、PI3K蛋白磷酸化，抑制Caspase-3、Bax蛋白表达，升高Bcl-2蛋白的表达，减轻海马神经元凋亡，从而改善高血压合并VCI大鼠的认知功能。
Abstract：Objective: To investigate the interventional effects of Tianma Gouteng Decoction on rats with spontaneous hypertension combined with vascular cognitive impairment model, and to observe the effects of this formula on phosphatidylinositol 3-kinase(PI3K)/serine/threonine kinase(AKT) signaling pathway, and to reveal its potential functional mechanism. Methods: Thirty-six male SHR rats were randomly grouped into sham operation group, model group, nimodipine group(9.375 mg/kg) and high, medium and low dose groups of Tianma Gouteng Decoction(23.750 g/kg, 11.875 g/kg and 5.938 g/kg) by strict randomized numerical table method. Six wistar rats were taken as a blank control group, and were treated with unilateral common carotid artery occlusion plus high salt diet. A rat model of spontaneous hypertension combined with vascular cognitive impairment was established by the method, followed by continuous treatment with nimodipine and different doses of Tianma Gouteng Decoction for 28d. Morris water maze test was used to test the escape latency, target quadrant residence time and the number of crossing platforms in each group of rats, Nissl staining was used to observe the pathological changes of neurons in rat hippocampal CA1 area and Nissl vesicles, TUNEL staining was used to evaluate the effect on neuronal apoptosis in rat hippocampal CA1 area, Western blotting was used to detect the protein expression of AKT, PI3K, p-AKT and p-PI3K in hippocampal tissues of each group of rats. Detection of protein expression and phosphorylation levels of AKT and PI3K in hippocampal tissues of rats in various groups by Western Blotting, and enzyme-linked immunosorbent assay(ELISA) was used to detect the protein expression of Caspase-3, Bcl-2 and Bax in the hippocampal tissues of rats in each group as well as the protein expression of ET-1 in serum.Results: Compared with the blank group and sham operation, rats in the model group had increased blood pressure, prolonged evasion latency, decreased target quadrant residence time, and decreased number of crossing platforms(P<0.01); hippocampal expression of Caspase-3 and Bax proteins increased significantly(P<0.01), and expression of Bcl-2 proteins decreased(P<0.01); hippocampal cell counts in the model group decreased(P<0.01), decrease in Nitrosomes, increase in TUNEL-positive cells(P<0.01), and significant decrease in phosphorylation levels of AKT and PI3K proteins(P<0.01).Compared with the model group, rats in the high, medium and nimodipine dose groups of Tianma Gouteng Decoction and the model group had lower blood pressure, reduced evasion latency, prolonged residence time in the target quadrant, and increased number of crossings of the plateau(P<0.01); protein expression of Caspase-3 and Bax decreased in the high, medium and low dose groups of Tianma Gouteng Decoction and the nimodipine group(P<0.01) and Bcl-2 protein expression rose(P<0.01); hippocampal cell counts increased(P<0.01), nidus bodies increased, and TUNEL-positive cells decreased(P<0.01) in the rat. protein expression rose(P<0.01); hippocampal cell counts increased in rats(P<0.01), Nitrosomes increased, and TUNEL-positive cells decreased(P<0.01); and AKT and PI3K protein phosphorylation levels were significantly elevated(P<0.01). Conclusion: Tianma Gouteng Decoction may improve the cognitive function of SHR combined with VCI rats by promoting the phosphorylation of AKT and PI3K proteins, inhibiting the expression of Caspase-3 and Bax proteins, elevating the expression of Bcl-2 proteins, and attenuating the apoptosis of hippocampal neurons.
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基本信息:
DOI：10.13210/j.cnki.jhmu.20241031.002
中图分类号:R285.5
引用信息:
[1]张红晓,李纳川,赵凡莹,等.天麻钩藤饮通过PI3K/AKT信号通路介导细胞凋亡改善高血压合并血管性认知障碍作用机制[J].海南医科大学学报,2025,31(15):1141-1152.DOI:10.13210/j.cnki.jhmu.20241031.002.
基金信息:
北京中医药大学第三附属医院院级培育（BZYSY-2022-PYMS-10）~~
2024-11-01
2024-11-01
2024-11-01