复方七芍降压片对SHR大鼠内皮细胞线粒体自噬和炎症应激的影响

Effect of compound qishao hypotensive tablet on mitochondrial autophagy and inflammatory response in SHR endothelial cells

目的分析复方七芍降压片对原发性高血压大鼠(SHR)内皮细胞线粒体自噬和炎症应激的影响。方法将50只SHR大鼠均分为空白组[予10 mg/(kg·d)生理盐水灌胃]、阳性对照组[予10 mg(kg·d)厄贝沙坦灌胃]及复方七芍(低、中、高)剂量组[分别予5、10、20 mg/(kg·d)复方七芍降压片灌胃],1次/d、连续30 d;麻醉处死大鼠并取腹主动脉组织检测PINK1蛋白、Parkin蛋白、LC3蛋白、丙二醛(MDA)、白细胞介素-1β(IL-1β)、肿瘤坏死因子-α(TNF-α)、一氧化氮(NO)、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)水平;以高剂量组PINK1蛋白、Parkin蛋白、LC3蛋白含量为因变量,以MDA、SOD、CAT、IL-1β、TNF-α、NO含量为自变量,分别拟合回归方程分析自变量对因变量的综合影响。结果阳性对照组、复方七芍各剂量组大鼠腹主动脉组织内皮细胞PINK1、Parkin、LC3、MDA、IL-1β、TNF-α以及NO含量高于空白组,复方七芍各剂量组低于阳性对照组,其中复方七芍低剂量>中剂量组>高剂量组(P<0.05);阳性对照组、复方七芍各剂量组大鼠腹主动脉组织内皮细胞SOD、CAT含量低于空白组,复方七芍各剂量组高于阳性对照组,其中低剂量组<中剂量组<高剂量组(P<0.05);对照组、阳性对照组、复方七芍各剂量组大鼠PINK1蛋白、Parkin蛋白、LC3蛋白含量与MDA、CAT、IL-1β、TNF-α、NO含量呈正相关,与SOD、CAT含量呈负相关(P<0.05);MDA、SOD、CAT、IL-1β、TNF-α、NO含量对PINK1、Parkin、LC3蛋白含量的综合影响均有统计学意义(P<0.05)。结论复方七芍降压片能明显改善SHR大鼠内皮细胞的炎症应激状态,其机制可能与影响线粒体自噬有关。

Objective To analyze the effect and correlation of compound qishao hypotensive tablets on mitochondrial autophagy and inflammatory stress state of endothelial cells in spontaneously hypertensive rats(SHRs).Methods Fifty SHRs were divided into blank group[intragastric administration of 10(mg/kg·d)normal saline],positive control group[10(mg/kg·d)irbesartan],low-,medium-,and high-dose compound qishao groups[5,10,and 20(mg/kg·d)of compound qishao hypotensive tablet,respectively].The treatment was once/d for 30 consecutive days.The rats were sacrificed using anesthesia.Abdominal aorta tissues were taken to detect the protein expression levels of PINK1,Parkin and LC3,malondialdehyde(MDA),interleukin-1β(IL-1(),tumor necrosis factor-((TNF-(),the levels of nitric oxide(NO),superoxide dismutase(SOD),and catalase(CAT).The protein levels of PINK1,Parkin,and LC3 in high-dose group were taken as dependent variables,while MDA,SOD,CAT,IL-1β,TNF-α,and NO were taken as independent variables.Fitting regression equation was used to analyze the comprehensive effect of independent variables on each dependent variable.Results PINK1,Parkin,LC3,MDA,IL-1β,TNF-α,and NO in the endothelial cells of abdominal aorta tissues were higher in positive control and compound qishao groups than those in blank group,and they were lower in compound qishao groups than those in positive group,with low-dose>medium-dose>high-dose compound qishao groups(P<0.05).The contents of SOD and CAT in the endothelial cells of abdominal aorta tissues were less in positive and compound qishao groups than those in blank group,and they were greater in compound qishao groups than those in positive group,with low-dose<medium-dose<high-dose compound qishao groups(P<0.05).The protein levels of PINK1,Parkin,and LC3 were positively associated with the levels of MDA,CAT,IL-1(,TNF-α,and NO in control,positive control,and compound qishao groups,and negatively correlated with the levels of SOD and CAT(P<0.05).The comprehensive effects of MDA,SOD,CAT,IL-1(,TNF-α,and NO on PINK1,Parkin,and LC3 protein contents were statistically significant(P<0.05).Conclusion Compound qishao hypotensive tablet can significantly improve the inflammatory stress state of SHR endothelial cells,and its mechanism may be related to the effect of mitochondrial autophagy.