摘要
目的:基于核转录因子-κB(NF-κB)信号通路探究活血定痛方对心肌缺血再灌注损伤动物模型的保护作用。方法:SD大鼠随机分为假手术组、模型组、复方丹参滴丸组,活血定痛高、低剂量组,活血定痛高剂量+NF-κB抑制剂组,各组动物连续给药2周,活血定痛高剂量组+二硫代氨基甲酸吡咯烷(PDTC)组于造模后次日腹腔注射PDTC;结扎左冠状动脉前降支造模,检测左心功能及Na^+-K^+-ATP酶。各组动物腹主动脉取血,酶联免疫吸附测定(ELISA)法检测血清中肌酸激酶同工酶(CK-MB),肌钙蛋白T(c Tn T),肿瘤坏死因子-α(TNF-α),白细胞介素-6(IL-6),白细胞介素-1β(IL-1β),超氧化物歧化酶(SOD),丙二醛(MDA),谷胱甘肽过氧化物酶(GSH-Px)。蛋白免疫印迹法(Western blot)检测心肌NF-κB,NF-κB抑制蛋白α(IκBα),NF-κB抑制蛋白激酶(IκκB)的蛋白表达。结果:与模型组比较,复方丹参滴丸组和活血定痛高、低剂量组可不同程度降低血清中CK-MB,c Tn T,TNF-α,IL-6,IL-1β,MDA的水平,增加SOD,GSH-Px水平,上调心肌组织Iκκ水平激酶IκBα和IκκB的蛋白表达水平,提高心肌缺血再灌注模型动物心肌能量代谢过程中的Na^+-K^+-ATP酶活性(P <0. 05);而活血定痛高剂量组+PDTC并未降低血清中CK-MB,c Tn T,TNF-α,IL-6,IL-1β,MDA的含量,增加SOD,GSH-Px水平,上调心肌组织Iκκ水平激酶IκBα和IκκB的蛋白表达水平,与模型组比较差异无统计学意义。结论:活血定痛方可通过上调心肌缺血再灌注模型心肌组织中IκBα和IκκB的蛋白表达,抑制NF-κB信号通路的活化,降低炎症介质表达及自由基产生,提高心肌能量代谢过程中的Na^+-K^+-ATP酶活性,达到较好的抗心肌缺血再灌注损伤的作用。
Objective: To explore the protective effect of Huoxue Dingtong prescription on myocardial ischemia reperfusion injury in animal model based on nuclear transcription factor-κB( NF-κB) signal pathway.Method: In compound Danshen dropping pill group,SD rats were randomly divided into sham group,model group,compound salvia miltiorrhiza dropping pill group,high-dose Huoxue Dingtong group low-dose Huoxue Dingtong group,high-dose Huoxue Dingtong + NF-κB inhibitor group. The rats in each group were administered continuously for 2 weeks. The rats in high-dose Huoxue Dingtong group + pyrrolidine dithiocarbamate( PDTC)group were intraperitoneally administered the next day after modeling. Injection with PDTC and ligation of anterior descending branch of left coronary artery were performed to detect left ventricular function and Na^+-K^+-ATPase activity. Blood was collected from each animal abdominal aorta,and enzyme-linked immunosorbent assay( ELISA)was used to detect serum creatine kinase isoenzyme( CK-MB),troponin T( cTnT),tumor necrosis factor-α( TNF-α),interleukin-6( IL-6),interleukin-1β( IL-1β),superoxide gasification enzyme( SOD),malondialdehyde( MDA),glutathione peroxidase( GSH-Px). Western blot was used to detect the expressions of NF-κB,NF-κB inhibitor α( IκBα) and IκB kinase( IκκB) in myocardium. Result: Compared with model group,compound Danshen dropping pill group,high-dose Huoxue Dingtong group and low-dose Huoxue Dingtong group could reduce serum CK-MB,cTnT,TNF-α,IL-6,IL-1β,MDA,increase SOD and GSH-Px contents,increase the protein expressions of IκBα and IκB in myocardial tissue,and increase the activity of Na^+-K^+-ATPase in myocardial energy metabolism in myocardial ischemia-reperfusion model rats( P < 0. 05). However,high-dose Huoxue Dingtong group + PDTC did not decrease serum CK-MB,c Tn T,TNF-α,IL-6,IL-1β and MDA,increase SOD,GSH-Px,and increase the protein expression levels of IκBα and IκκB in myocardial tissue. There was no significant difference between high-dose Huoxue Dingtong group + PDTC and model group. Conclusion: Huoxue Dingtang prescription can inhibit the activation of NF-κB signaling pathway,reduce the expression of inflammatory mediators and the production of free radicals,and increase the activity of Na^+-K^+-ATPase in the process of myocardial energy metabolism by upregulating the expressions of IκBα and IκκB proteins in myocardial tissue of myocardial ischemia-reperfusion model.
作者
周玥
钟杭
李成朋
刘冰
ZHOU Yue;ZHONG Hang;LI Cheng-peng;LIU Bing(School of Pharmacy,Guizhou University,Guiyang 550025,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2020年第6期39-45,共7页
Chinese Journal of Experimental Traditional Medical Formulae
基金
贵州省科技计划项目(黔科合基础[2017]1061号,黔科合基础[2019]1121号)。
关键词
复方丹参滴丸
活血定痛方
心肌缺血再灌注
核转录因子-κB(NF-κB)
compound Danshen dropping pills
Huoxue Dingtong prescription
myocardial ischemia reperfusion
nuclear transcription factor-κB (NF-κB)
