Dong Aiai, Cui Yiyi, Wang Weifeng, Yang Duanrong, Wu Ronglin, Wang Wenrui, Han Wenhua, Lai Xiaoyun, Wang Haijun, Liu Haixin, Xue Ling, Ji Laixi
Second Clinical College of Shanxi University of CM, Jinzhong 030619, China.
Baoding First Hospital of TCM.
Zhongguo Zhen Jiu. 2024 Aug 12;44(8):931-8. doi: 10.13703/j.0255-2930.20240204-0001.
To observe the effects of acupuncture at "antihypertensive acupoint prescription" on endothelial active factors and related autonomic neurotransmitters in spontaneous hypertension rats, and explore the vascular regulation and central regulation mechanisms of acupuncture for anti-hypertension.
Thirty SPF grade male spontaneous hypertension rats were randomly divided into a model group (15 rats) and an acupuncture group (15 rats). Besides, 15 Wistar Kyoto rats were collected as a blank control group (normal group). In the acupuncture group, acupuncture was delivered at the "antihypertensive acupoint prescription" (bilateral "Renying" [ST 9], "Quchi" [LI 11], "Zusanli" [ST 36], "Taichong" [LR 3] and "Neiguan" [PC 6]), with needles retained for 30 min, once daily. The duration of intervention was 28 days. Every week, using the the irritation scale, the sign of sympathetic irritation was evaluated dynamically. The arterial blood pressure of the rats tail was determined, using non-invasive blood pressure measurement system. ELISA was adopted to detect the levels of calcitonin gene-related peptide (CGRP), nitric oxide (NO), endothelin-1 (ET-1), neuropeptide Y (NPY) in the serum. DAB chromogenic in situ hybridization (CISH) was provided to detect the mRNA expression of endothelial nitric oxide synthase (eNOS) in the internal carotid artery and the arcuate nucleus (ARC), and that of CGRP in the paraventricular nucleus posterior (PVP) and the ventrolateral medulla (VLM). Liquid chromatography-mass spectrometry (LC-MS) was used to detect the levels of epinephrine (E) and norepinephrine (NE) in the paraventricular nucleus anterior (PVA).
Compared with the normal group, the irritation scores, systolic blood pressure and diastolic blood pressure were increased at each time point in the model group (<0.05). When compared with the model group, the irritation scores after the intervention for 3 and 4 weeks, and systolic and diastolic blood pressure after intervention for 2, 3 and 4 weeks were reduced in the acupuncture group (<0.05). In comparison with the normal group, the serum CGRP and NO levels of the rats were decreased (<0.05), and the serum ET-1 and NPY levels, as well as E and EN levels in PVA were increased (<0.05) in the model group. The levels of serum CGRP and NO were elevated (<0.05), and the serum ET-1 and NPY levels, as well as E and EN levels of PVA were reduced (<0.05) in the acupuncture group when compared with those of the model group. In the model group, the media of internal carotid artery exhibited thickening and remodeling, while the neuron volume in ARC was small. In the acupuncture group, every layer of internal carotid artery was acceptably arranged, and the parvicellular neuron of ARC was moderate in volume. For the in situ hybridization of eNOS mRNA for the rats of each group, the smooth muscle cells were predominantly expressed in each layer of the internal carotid artery, whereas the expression of parvicellular neurons was dominated in ARC. In the model group, the large and small neurosecretory cells were distributed sparsely in the nerves of PVP; in the acupuncture group, the cells of these two species were distributed regularly; and there were few species of glial cell in the VLM of either the model group or the acupuncture group. In each group, for the in situ hybridization of CGRP mRNA, the small neurosecretory cells were expressed predominately in the PVP, while, the expression of glial cell nuclei and the cell cytoplasm was dominated in the VLM. Compared with the normal group, the mRNA expression of eNOS in the internal carotid artery and ARC and that of CGRP mRNA in the PVP and VLM was decreased in the model group (<0.05). In the acupuncture group, when compared with the model group, the mRNA expression of eNOS in the internal carotid artery and ARC and that of CGRP in the PVP and VLM was increased in the acupuncture group (<0.05).
Acupuncture at "antihypertensive acupoint prescription" can upregulate the level of vascular relaxing factors, downregulate the level of contracting factors, enhance the response of relaxing factors in targeting blood vessels and regulating the center. The mechanism may be related to the modulation of the sympathetic-adrenergic autonomic neurotransmitters in the paraventricular nucleus in spontaneous hypertension rats.
观察针刺“降压穴组方”对自发性高血压大鼠血管内皮活性因子及相关自主神经递质的影响，探讨针刺抗高血压的血管调节及中枢调节机制。
将30只SPF级雄性自发性高血压大鼠随机分为模型组（15只）和针刺组（15只）。另取15只Wistar Kyoto大鼠作为空白对照组（正常组）。针刺组针刺“降压穴组方”（双侧“人迎”[ST 9]、“曲池”[LI 11]、“足三里”[ST 36]、“太冲”[LR 3]和“内关”[PC 6]），留针30分钟，每日1次。干预时间为28天。每周动态评估交感神经兴奋体征，采用无创血压测量系统测定大鼠尾动脉血压。采用ELISA法检测血清中降钙素基因相关肽（CGRP）、一氧化氮（NO）、内皮素-1（ET-1）、神经肽Y（NPY）水平。采用DAB显色原位杂交（CISH）法检测颈内动脉和弓状核（ARC）中内皮型一氧化氮合酶（eNOS）的mRNA表达，以及室旁核后部（PVP）和延髓腹外侧区（VLM）中CGRP的mRNA表达。采用液相色谱-质谱联用（LC-MS）法检测室旁核前部（PVA）中肾上腺素（E）和去甲肾上腺素（NE）水平。
与正常组比较，模型组各时间点兴奋评分、收缩压和舒张压均升高（<0.05）。与模型组比较，针刺组干预3、4周后兴奋评分，干预2、3、4周后收缩压和舒张压均降低（<0.05）。与正常组比较，模型组大鼠血清CGRP和NO水平降低（<0.05），血清ET-1和NPY水平以及PVA中E和NE水平升高（<0.05）。与模型组比较，针刺组血清CGRP和NO水平升高（<0.05），血清ET-1和NPY水平以及PVA中E和NE水平降低（<0.05）。模型组颈内动脉中膜增厚、重塑，ARC神经元体积较小。针刺组颈内动脉各层排列尚可，ARC小细胞神经元体积适中。对各组大鼠进行eNOS mRNA原位杂交，颈内动脉各层平滑肌细胞为主表达，ARC中小细胞神经元为主表达。模型组PVP神经中大、小神经分泌细胞分布稀疏；针刺组这两种细胞分布规则；模型组和针刺组VLM中胶质细胞种类均较少。各组进行CGRP mRNA原位杂交，PVP中小神经分泌细胞为主表达，VLM中胶质细胞核和细胞质为主表达。与正常组比较，模型组颈内动脉和ARC中eNOS mRNA表达以及PVP和VLM中CGRP mRNA表达降低（<0.05）。与模型组比较，针刺组颈内动脉和ARC中eNOS mRNA表达以及PVP和VLM中CGRP mRNA表达升高（<0.05）。
针刺“降压穴组方”可上调血管舒张因子水平，下调收缩因子水平，增强靶血管舒张因子反应及中枢调节作用。其机制可能与调节自发性高血压大鼠室旁核交感-肾上腺素能自主神经递质有关。
