活血散风针刺法对高血压失眠大鼠作用机制研究

Study on Mechanism of Huoxue Sanfeng Acupuncture in Hypertensive Insomnia Rats

摘要:

目的：观察活血散风针刺法对高血压失眠大鼠的作用机制。方法：将40只自发性高血压大鼠（简称SHR大鼠） 随机分为空白组、模型组、针刺组、西药组，每组10只。采用对氯苯丙氨酸（PCPA） 构建高血压失眠大鼠模型。针刺组取穴双侧人迎、合谷、太冲，均施以平补平泻手法；西药组予佐匹克隆灌胃治疗，剂量根据人与大鼠体表面积公式计算；空白组、模型组均不予干预治疗，仅按针刺组相同方法麻醉大鼠。干预7天后，采用酶联免疫法检测下丘脑组织5-羟色胺（5-HT）、谷氨酸（Glu）、γ-氨基丁酸（GABA） 含量，采用Western Blot法检测谷氨酸脱羧酶1（GAD1） 及谷氨酰胺合成酶（GS） 的含量。结果：与空白组比较，模型组大鼠5-HT浓度下降，Glu浓度上升，GABA浓度下降，Glu/GABA比值上升，GAD1含量下降，GS蛋白含量上升，差异均有统计学意义（P＜0.05）。与模型组比较，针刺组和西药组5-HT 浓度上升，Glu 浓度下降，GABA浓度升高，Glu/GABA比值下降，差异均有统计学意义（P＜0.05），GAD1含量上升（P＜0.05），GS蛋白含量下降（P＜0.05）；与西药组比较，针刺组GAD1浓度上升，差异有统计学意义（P＜0.05）。结论：针刺组与西药组均对高血压失眠大鼠有治疗效果。活血散风针刺法能够有效改善SHR失眠大鼠症状，其作用机制可能是通过提高下丘脑内5-HT浓度，调控Glu/GABA-谷氨酰胺（Gln） 代谢环路：提高GAD1表达水平，促进GABA生成，降低GS蛋白表达，减少Glu再生成，下调Glu/GABA比值。

Abstract:

Abstract：Objective：To observe the mechanism of Huoxue Sanfeng acupuncture on hypertensive insomnia rats. Methods：Forty SHR rats were randomly divided into the blank group，the model group，the acupuncture group，and the western medicine group，with 10 rats in each group. The hypertensive insomnia rat model was established using pchlorophenylalanine （PCPA）. In the acupuncture group， bilateral points Renying （ST9）， Hegu （LI4）， and Taichong（ LR 3） were selected and treated with a uniform reinforcing and reducing method. The western medicine group received intragastric treatment with Zopiclone，with the dose calculated based on the human-to-rat body surface area ratios. No interventions were applied to either the blank or model groups， while all rats in the two groups were anesthetized similarly to those in the acupuncture group. After seven days of intervention， the levels of 5- hydroxytryptamine （5-HT）， glutamic acid （Glu）， and γ -aminobutyric acid （GABA） in the hypothalamus were detected by enzyme-linked immunosorbent assay （ELISA）. The levels of glutamate decarboxylase 1 （GAD1） and glutamine synthetase （GS） were detected by Western Blot. Results：Compared with the blank group，the model group showed a decrease in 5-HT and GABA concentration，an increase in Glu concentration，an increase in the Glu/GABA ratio， a decrease in GAD1 content， and an increase in GS protein content； all differences were statistically significant （P＜0.05）. Compared with the model group，the acupuncture group and western medicine group showed an increase in 5-HT concentration，a decrease in Glu concentration，an increase in GABA concentration，and a decrease in the Glu/GABA ratio； all differences were statistically significant （P＜0.05）. In addition， GAD1 content increased （P＜0.05） and GS protein content decreased （P＜0.05） in both treatment groups compared with the model group. Compared with the western medicine group，the acupuncture group showed an increase in GAD1 concentration， with a statistically significant difference （P＜0.05）. Conclusion： Both the acupuncture group and the western medicine group had therapeutic effects on hypertensive insomnia rats. Huoxue Sanfeng acupuncture can effectively improve the symptoms of SHR insomnia rats. Its mechanism may involve increasing the 5-HT concentration in the hypothalamus and regulating the Glu/GABA-Gln metabolic loop： upregulating GAD1 expression to promote GABA production， downregulating GS protein expression to reduce Glu regeneration， and thereby lowering the Glu/GABA ratio.