摘要
目的:基于内皮型一氧化氮合酶(endothelial nitric oxide synthase,eNOS)/一氧化氮(nitric oxide,NO)信号通路探讨脑清通颗粒对肝热痰瘀证高血压大鼠的降压作用及机制。方法:随机从70只6周龄SPF级雄性SD大鼠中选取10只为正常组,其余60只大鼠采用皮下注射环孢菌素A加灌胃给予附子和高脂乳剂制备肝热痰瘀证高血压大鼠模型。取造模成功的大鼠50只,随机分为模型组、卡托普利组及脑清通颗粒低(1.3 g·kg^(-1))、中(2.6 g·kg^(-1))、高(5.2 g·kg^(-1))剂量组,每组10只,于造模成功的第2天开始给予相应药物灌胃,正常组和模型组灌胃给予等体积生理盐水,每日1次,连续21 d。分别于造模前、造模第7天、造模第14天、给药第7天、给药第14天和给药第21天测量大鼠尾动脉压,计算平均动脉压(mean arterial pressure,MAP);HE染色观察血管内膜形态学变化;ELISA检测血清血管紧张素Ⅱ(angiotensinⅡ,AngⅡ)、内皮素-1(endothelin-1,ET-1)、eNOS、NO含量;免疫组化和Western Blot检测胸主动脉组织中eNOS蛋白表达水平。结果:与正常组比较,模型组MAP及血清ET-1、AngⅡ水平明显升高,血清NO、eNOS水平及胸主动脉eNOS蛋白表达水平显著降低;与模型组比较,脑清通颗粒各剂量组血清ET-1、AngⅡ水平显著降低,NO、eNOS水平显著升高,胸主动脉eNOS蛋白表达水平显著升高,给药14 d后MAP显著下降,差异均具有统计学意义(P<0.05)。HE染色显示,模型组大鼠血管内膜粗糙,结构不完整,有明显脱落现象,平滑肌细胞数量明显减少,而脑清通颗粒各剂量组大鼠血管内膜损伤均得到了不同程度的改善。结论:脑清通颗粒能改善肝热痰瘀证高血压模型大鼠血管内膜损伤,降低血压,其机制可能与抑制ET-1和AngⅡ的释放,激活eNOS/NO信号通路有关。
Objective:Based on endothelial nitric oxide synthase(eNOS)/nitric oxide(NO)signaling pathway,to investigate the blood pressure of Naoqingtong Granule on hypertensive rats with liver-heat-phlegm-stasis syndrome action and mechanism.METHODS:106-week-old SPF male SD rats were randomly selected as normal group,and the remaining 60 rats were subcutaneously injected with cyclosporine A plus intragastric administration of aconite and high-fat emulsion to prepare Liver Heat and Phlegm Stasis in Hypertensive model rat.Fifty rats with successful modeling were selected and randomly divided into model group,captopril group and Naoqingtong Granules low(1.3 g·kg^(-1)),medium(2.6 g·kg^(-1)),high(5.2 g·kg^(-1))dose group,10 animals in each group,were given the corresponding drugs by gavage on the second day after the successful modeling,and the normal group and model group were given the same volume of normal saline by gavage,once a day,for 21 consecutive days.Before modeling,modeling day 7,modeling day 14,administration day 7,administration day 14,and administration day 21,the rat tail artery pressure was measured,and the mean arterial pressure was calculated.(MAP);HE staining was used to observe the morphological changes of the vascular intima;ELISA was used to detect the content of serum angiotensin II(angiotensin II,Ang II),endothelin-1(endothelin-1,ET-1),eNOS,NO;Western Blot detected the expression level of eNOS protein in thoracic aorta tissue.Results:Compared with the normal group,the model group significantly increased the levels of MAP,serum ET-1 and AngⅡ,while the serum NO,eNOS levels and thoracic aorta eNOS protein expression levels were significantly decreased.Serum ET-1 and AngⅡlevels were significantly decreased,NO and eNOS levels were significantly increased,thoracic aorta eNOS protein expression levels were significantly increased,and MAP was significantly decreased after 14 days of administration,the differences were statistically significant(P<0.05).HE staining showed that the intima of the blood vessels of the rats in the model group was rough,the structure was incomplete,there was obvious shedding phenomenon,and the number of smooth muscle cells was significantly reduced.Conclusion:Naoqingtong Granules can improve the vascular intima and reduce blood pressure in hypertensive rats with Liver Heat and Phlegm Stasis syndrome,and its mechanism may be related to inhibiting the release of ET-1 and Ang II and activating the eNOS/NO signaling pathway.
作者
严亚锋
白海侠
周海哲
YAN Yafeng;BAI Haixia;ZHOU Haizhe(Shaanxi University of Traditional Chinese Medicine,Xianyang Shaanxi China 712046)
出处
《中医学报》
CAS
2022年第11期2390-2396,共7页
Acta Chinese Medicine
基金
陕西省科技厅课题项目(2020SF-341)
陕西省中医药管理局课题项目(2021-ZZ-JC043)
陕西张氏脑病学术流派传承工作室建设项目[陕中医药发(2018)40号]。
关键词
高血压
肝热痰瘀证
脑清通颗粒
eNOS/NO信号通路
大鼠
hypertension
Liver Heat and Phlegm Stasis syndrome
Naoqingtong Granules
eNOS/NO signaling pathway
rat
