健脾化浊方通过NOX4/ROS/NF-κB通路调节动脉粥样硬化大鼠主动脉炎症及氧化应激反应的研究

Effect of Jianpi Huazhuo Recipe on Inflammation and Oxidative Stress Response in Aorta of Atherosclerotic Rats by Regulating NOX/ROS-NF-κB Pathway

目的:研究健脾化浊方通过NAPDH氧化酶4(NOX4)/活性氧簇(ROS)/核因子-κB(NF-κB)通路调节动脉粥样硬化(AS)大鼠主动脉炎症及氧化应激反应的作用。方法:SD大鼠随机分为正常对照组、模型对照组、健脾化浊方10 g/kg组、20 g/kg组、40 g/kg组和健脾化浊方40 g/kg+内毒素(LPS)150μg/kg组,建立AS模型后灌胃给予不同剂量健脾化浊方或联合NF-κB激动剂LPS腹腔注射。检测主动脉粥样硬化的病理改变、气滞血瘀标志物及主动脉中炎症细胞因子、氧化应激指标、NOX4/ROS/NF-κB通路分子。结果:与模型对照组比较,健脾化浊方10 g/kg组、20 g/kg组、40 g/kg组大鼠的低切血粘度、中切血粘度、高切血粘度、血清中ET-1、GMP-140的含量及主动脉中肿瘤坏死因子-α(TNF-α)、白细胞介素(IL)-1β、细胞间黏附分子-1(ICAM-1)、C反应蛋白(CRP)、丙二醛(MDA)、晚期氧化蛋白产物(AOPP)、ROS的含量明显降低、NOX4、NF-κB的蛋白表达均明显下调,血清中一氧化氮(NO)、降钙素基因相关肽(CGRP)的含量及主动脉中总抗氧化力(T-AOC)的含量均明显升高;与健脾化浊方40 g/kg组比较,健脾化浊方40 g/kg+LPS 150μg/kg组大鼠的低切血粘度、中切血粘度、高切血粘度、血清中ET-1、GMP-140的含量及主动脉中TNF-α、IL-1β、ICAM-1、CRP、MDA、AOPP、ROS的含量明显增加、NOX4、NF-κB的蛋白表达量均明显上调,血清中NO、CGRP的含量及主动脉中T-AOC的含量明显降低。结论:健脾化浊方能够改善AS大鼠主动脉的AS病理改变及气滞血瘀表现,该作用与抑制NOX4/ROS/NF-κB通路介导的炎症反应及氧化应激反应有关。

Objective:To study the effect of Jianpi Huazhuo Recipe on regulating inflammation and oxidative stress response of aorta by NOX/ROS-NF-κB pathway in atherosclerosis(AS)rats.Methods:SD rats were randomly divided into the control group,AS model group,Jianpi Huazhuo groups(10 g/kg、20 g/kg、40 g/kg)and Jianpi Huazhuo(40 g/kg)+LPS(150μg/kg)group.After AS model was established,SD rats were orally given different doses of Jianpi Huazhuo Recipe or Jianpi Huazhuo combined with NF-κB agonist LPS(intraperitoneal injection).Pathological changes of aortic atherosclerosis were observed,Qizhixueyu markers,inflammation cytokines,oxidative stress index,NOX/ROS-NF-κB pathway molecules in aorta were detected.Results:Compared with AS group,the low shear blood viscosity,medium shear blood viscosity and high shear blood viscosity,the contents of ET-1,GMP-140 in serum and the contents of TNF-a,IL-1β,ICAM-1,CRP,MDA,AOPP,ROS in serum and the expressions of NOX4,NF-κB in aorta in three Jianpi Huazhuo Recipe groups were significantly decreased.The contents of NO,CGRP in serum and the content of T-AOC in aorta were significantly increased.Compared with 40 g/kg Jianpi Huazhuo group,the low shear blood viscosity,medium shear blood viscosity and high shear blood viscosity,the contents of ET-1,GMP-140 in serum and the contents of TNF-a,IL-1β,ICAM-1,CRP,MDA,AOPP,ROS in serum and the expressions of NOX4,NF-κB in 40 g/kg+LPS 150μg/kg group were significantly increased,while the contents of NO,CGRP in serum and the content of T-AOC in aorta significantly decreased.Conclusion:Jianpi Huazhuo recipe can improve pathological changes of aorta in AS and manifestations of Qizhixueyu syndrome in AS rats,this effect is related to inhibiting inflammation and oxidative stress mediated by NOX4/ROS-NF-κB pathway.