Lu Cheng-Xuan, Xu Qian, Ye Ru-Lin, Zhu Hai-Bin, Wu Huan-Xi, Zhang Jian-Bin
College of Acupuncture-moxibustion and Massage, College of Health and Rehabilitation, Nanjing University of Chinese Medicine, Nanjing 210023, China.
The Second Affiliated Hospital of Nanjing University of Chinese Medicine/The Second Chinese Medicine Hospital of Jiangsu Province, Nanjing 210017.
Zhen Ci Yan Jiu. 2023 Apr 25;48(4):331-8. doi: 10.13702/j.1000-0607.20220056.
To explore the antioxidant effect of moxibustion on vascular endothelial function and the under-lying mechanism.
Forty male SD rats were randomly divided into blank, model, moxibustion and endothelial nitric oxide synthase (eNOS) inhibitor groups, with 10 rats in each group. Hyperlipidemia rat model was established by high fat diet for 8 weeks. Rats in the moxibustion group received 45 ℃ moxibustion at "Zusanli" (ST36) for 10 min once daily for consecutive 4 weeks. Rats in the eNOS inhibitor group received intraperitoneal injection of eNOS inhibitor L-NAME (1 mg/100 g) at the same time of moxibustion intervention. The morphology of abdominal aorta endothelium was observed by HE staining. Lipid deposition in abdominal aorta was observed by oil red O staining. The contents of total cholesterol (TC), triglyceride (TG), high density lipoprotein cholesterol (HDL-C), low density lipoprotein cholesterol (LDL-C) in serum and reactive oxygen species (ROS), nitric oxide (NO), superoxide dismutase (SOD), oxidized LDL lipoprotein (ox-LDL), endothelin-1 (ET-1), eNOS, malondialdehyde (MDA) in serum and abdominal aorta were determined by ELISA. The expression of eNOS in abdominal aorta was detected by immunofluorescence.
HE staining of the abdominal aorta showed no significant pathological abnormality in the blank group; the endovascular cortex was rough, and the inner, media and outer membrane were rough in the model group; the nucleus and surrounding tissue structure were clear and the vascular wall was smooth in the moxibustion group; abdominal aorta texture was rough in the eNOS inhibitor group. Compared with the blank group, the area of oil red O staining in abdominal aorta increased (<0.05); the contents of serum TC, TG and LDL-C increased (<0.01, <0.05) while HDL-C decreased (<0.05); the contents of ET-1 in serum and abdominal aorta were increased (<0.01, <0.05) while the contents of NO and eNOS were decreased (<0.05, <0.001); the contents of ROS, ox-LDL and MDA in serum and abdominal aorta were increased (<0.001, <0.01, <0.000 1) while the content of SOD in abdominal aorta was decreased (<0.000 1); the expression level of eNOS in abdominal aorta was decreased (<0.05) in the model group. Compared with the model group, the area of oil red O staining in abdominal aorta decreased (<0.05); the contents of TC, TG and LDL-C in serum decreased (<0.05) while HDL-C increased (<0.05); the contents of ET-1 in serum and abdominal aorta were decreased (<0.01, <0.05) while the contents of NO and eNOS in abdominal aorta were increased (<0.001, <0.01); the contents of ROS and MDA in serum and abdominal aorta were decreased (<0.001, <0.01, <0.05), the content of ox-LDL was decreased (<0.01) and the content of SOD was increased (<0.000 1) in abdominal aorta; the expression level of eNOS in abdominal aorta was increased (<0.05) in the moxibustion group. Compared with the moxibustion group, the contents of serum TC, LDL-C and MDA in the eNOS inhibitor group were increased (<0.05); the contents of ET-1, ROS, ox-LDL and MDA in abdominal aorta were increased (<0.05), the contents of NO, eNOS and SOD were decreased (<0.05); the expression level of eNOS in abdominal aorta was decreased (<0.05).
45 ℃ moxibustion at ST36 can protect and repair vascular endothelial injury in abdominal aorta of hyperlipidemia rats and improve the oxidative stress of vascular endothelium.
探讨艾灸对血管内皮功能的抗氧化作用及其潜在机制。
将40只雄性SD大鼠随机分为空白组、模型组、艾灸组和内皮型一氧化氮合酶（eNOS）抑制剂组，每组10只。采用高脂饮食8周建立高脂血症大鼠模型。艾灸组大鼠每日于“足三里”（ST36）穴进行45℃艾灸10分钟，连续4周。eNOS抑制剂组大鼠在艾灸干预的同时腹腔注射eNOS抑制剂L-NAME（1mg/100g）。采用HE染色观察腹主动脉内皮形态。采用油红O染色观察腹主动脉脂质沉积。采用ELISA法检测血清中总胆固醇（TC）、甘油三酯（TG）、高密度脂蛋白胆固醇（HDL-C）、低密度脂蛋白胆固醇（LDL-C）以及血清和腹主动脉中活性氧（ROS）、一氧化氮（NO）、超氧化物歧化酶（SOD）、氧化型低密度脂蛋白（ox-LDL）、内皮素-1（ET-1）、eNOS、丙二醛（MDA）的含量。采用免疫荧光法检测腹主动脉中eNOS的表达。
腹主动脉HE染色显示，空白组未见明显病理异常；模型组血管内皮粗糙，内膜、中膜和外膜均粗糙；艾灸组细胞核及周围组织结构清晰，血管壁光滑；eNOS抑制剂组腹主动脉质地粗糙。与空白组比较，模型组腹主动脉油红O染色面积增加（<0.05）；血清TC、TG和LDL-C含量升高（<0.01，<0.05），HDL-C含量降低（<0.05）；血清和腹主动脉中ET-1含量升高（<0.01，<0.05），NO和eNOS含量降低（<0.05，<0.001）；血清和腹主动脉中ROS、ox-LDL和MDA含量升高（<0.001，<0.01，<0.0001），腹主动脉中SOD含量降低（<0.0001）；模型组腹主动脉中eNOS表达水平降低（<0.05）。与模型组比较，艾灸组腹主动脉油红O染色面积减小（<0.05）；血清TC、TG和LDL-C含量降低（<0.05），HDL-C含量升高（<0.05）；血清和腹主动脉中ET-1含量降低（<0.01，<0.05），腹主动脉中NO和eNOS含量升高（<0.001，<0.01）；血清和腹主动脉中ROS和MDA含量降低（<0.001，<0.01，<0.05），腹主动脉中ox-LDL含量降低（<0.01），SOD含量升高（<0.0001）；艾灸组腹主动脉中eNOS表达水平升高（<0.05）。与艾灸组比较，eNOS抑制剂组血清TC、LDL-C和MDA含量升高（<0.05）；腹主动脉中ET-1、ROS、ox-LDL和MDA含量升高（<0.05），NO、eNOS和SOD含量降低（<0.05）；腹主动脉中eNOS表达水平降低（<0.05）。
艾灸“足三里”穴45℃可保护和修复高脂血症大鼠腹主动脉血管内皮损伤，改善血管内皮氧化应激。
