针刺对高血压大鼠延髓头端腹外侧区氧化应激水平影响的研究

王雪蕊 郭玉红 徐霄龙 白云静 刘清泉

摘要 目的：從自由基来源和清除的角度探讨针刺对自发性高血压大鼠（SHR）延髓头端腹外侧区（RVLM）中氧化应激水平影响的潜在机制。方法：15只Wistar（WKY）大鼠为正常组，45只SHR大鼠随机分为高血压组、针刺组和非穴组。针刺组取“太冲穴”直刺，非穴组取非穴位浅刺，治疗1次/d，共14 d。采用遥测技术检测针刺对血压的影响;采用DHE染色检测针刺对RVLM中活性氧族（ROS）变化的影响;采用化学发光法检测针刺对RVLM中超氧化物歧化酶（SOD）、一氧化氮合酶（NOS），和NADPH氧化酶活性的影响;采用Western Blot检测针刺对RVLM中NADPH氧化酶、SOD、NOS各亚基或亚型蛋白表达的影响。结果：与正常组比较，高血压组大鼠收缩压显著增高（P<0.05），RVLM中ROS产生增多（P<0.05），NADPH氧化酶活性增高（P<0.05），其亚基gp91phox、p22phox和nNOS蛋白表达水平显著升高（P<0.05）。与高血压组和非穴组比较，针刺干预可显著下调SHR大鼠的收缩压（P<0.05），降低RVLM中ROS的水平（P<0.05），下调NADPH氧化酶活性（P<0.05）。各组中SOD和NOS活性，SOD1、SOD2和iNOS蛋白表达水平无显著改变。结论：针刺可显著改善SHR大鼠血压，降低RVLM中ROS的水平，其机制可能与下调NADPH氧化酶亚基gp91phox、p22phox、及NOS亚型nNOS的表达水平相关。

关键词 高血压;针刺;氧化应激;延髓头端腹外侧区;大鼠

Effects of Acupuncture on Oxidative Stress in the Rostral Ventrolateral Medulla of Spontaneously Hypertensive Rats

WANG Xuerui，GUO Yuhong ，XU Xiaolong，BAI Yunjing，LIU Qingquan

（Beijing Hospital of Traditional Chinese Medicine affiliated to Capital Medical University，Beijing 100029，China）

Abstract Objective：To explore the potential mechanism of acupuncture on oxidative stress level in the rostral ventrolateral medulla （RVLM） of spontaneously hypertensive rats （SHR） from the perspective of free radical source and elimination.Methods：A total of 15 Wistar （WKY） rats were in the normal group，and 45 SHR rats were randomly divided into a model group，an acupuncture group and a non-acupoint group.In the acupuncture group，“Taichong” acupoints were used for treatment once a day for 14 days.In the non-acupoint group received superficial needling was used.The effects of acupuncture on blood pressure were measured by telemetry; the effects of acupuncture on the changes of reactive oxygen species （ROS） in RVLM were detected by DHE staining; the effects of acupuncture on the activities of superoxide dismutase （SOD），nitrogen synthase （NOS） and NADPH oxidase activity in RVLM were detected by chemiluminescence; and the effects of acupuncture on the subunits or subunits expression of NADPH oxidase，SOD and NOS in RVLM were detected by Western Blot.Results：Compared with the normal group，the systolic blood pressure of the model group was significantly increased （P<0.05），ROS production in RVLM increased （P<0.05），the NADPH oxidase activity increased （P<0.05），and the protein expression levels of gp91phox，p22phox and nNOS were significantly increased （P<0.05）.com

pared with the model group and non-acupoint group，acupuncture intervention could significantly down-regulate the systolic blood pressure of SHR rats （P<0.05），the level of ROS in RVLM （P<0.05），and the activity of NADPH oxidase （P<0.05）.There was no significant change in the SOD and NOS activities，the protein expression of SOD1，SOD2 and iNOS among each groups.Conclusion：Acupuncture could significantly reduce high blood pressure and ROS levels in the RVLM of SHR rats.The mechanism may be related to the down-regulation of the expression of NADPH oxidase subunits gp91phox，p22phox and NOS subtype nNOS.

Keywords Hypertension; Acupuncture; Oxidative stress; Rostral ventrolateral medulla; Rats

中图分类号：R245文献标识码：Adoi：10.3969/j.issn.1673-7202.2021.01.012

高血压病是最为常见的心血管疾病之一，目前我国约有患者2亿人，其造成的靶器官损害严重影响了人民群众的健康[1]。既往研究发现，氧化应激是影响高血压发生发展和导致靶器官损伤的关键因素[2-3]。生理状态下，体内氧化和抗氧化系统维持动态平衡，防止活性氧族（ROS）对机体的损害。在高血压的病理状态下，机体或细胞内氧自由基的产生和消除失衡导致ROS在过量蓄积，从而引起氧化应激。ROS的主要来源包括还原型烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶，其他来源包括一氧化氮合成酶脱偶联（NOS）等。机体各组织包括血清中也包含有一定活性的抗氧化酶系，可及时有效地清除ROS。其中，超氧化物歧化酶（SOD）是体内含量最为广泛的抗氧化剂，可以通过清除自由基防止靶器官的氧化损伤。……