橙皮素通过激活SIRT1/NRF2信号改善阿霉素诱导的H9c2细胞毒性  

Hesperetin Alleviates Doxorubicin-Induced Cytotoxicity in H9c2 Cells by Activating SIRT1/NRF2 Signaling

作　　者：李新华[1] 闫爱丽 常晋瑞[3] 李汾[1] 朱娟霞[3] LI Xinhua;YAN Aili;CHANG Jinrui;LI Fen;ZHU Juanxia(Department of Pharmacology and Toxicology,Xi'an Medical University,Xi'an 710021,China;Department of Pathology,Xi'an Medical University,Xi'an 710021,China;Department of Physiology,Xi'an Medical University,Xi'an 710021,China)

机构地区：[1]西安医学院药理学与毒理学教研室,西安710021 [2]西安医学院病理学教研室,西安710021 [3]西安医学院生理学教研室,西安710021

出　　处：《四川大学学报（医学版）》2023年第5期947-953,共7页Journal of Sichuan University(Medical Sciences)

基　　金：陕西省教育厅专项科研计划项目(No.20JK0883、No.20JS135)资助。

摘　　要：目的本研究旨在探讨橙皮素(hesperetin,Hes)能否通过调控沉默信息调节因子1(silent information regulator 1,SIRT1)/核转录因子E2相关因子2(nuclear transcription factor E2-related factor 2,NRF2)信号减轻氧化应激改善阿霉素(doxorubicin,DOX)诱导的H9c2心肌细胞毒性。方法采用DOX诱导的H9c2心肌细胞毒性模型,随机分为4组:对照组(Control)、DOX处理组(DOX)、Hes加DOX处理组(DOX+Hes)以及Hes加SIRT1抑制剂EX527联合DOX处理组(DOX+Hes+EX527)。光镜下观察细胞形态,CCK-8检测各组细胞活力,流式细胞术检测各组细胞凋亡率,DCFH-DA染色观察各组ROS水平,ELISA试剂盒检测乳酸脱氢酶(lactic dehydrogenase,LDH)、超氧化物歧化酶(superoxide dismutase,SOD)、过氧化氢酶(catalase,CAT)和SIRT1活性以及丙二醛(malondialdehyde,MDA)含量,Western blot检测裂解的caspase-3(cleaved caspase-3)、细胞色素c(cytochrome c)、SIRT1、Ac-FOXO1、NRF2和血红素加氧酶1(heme oxygenase1,HO-1)的表达。结果和Control组相比,DOX组细胞形态肿胀,密度减小,细胞活力降低,培养基中LDH活性增加(P<0.01);细胞凋亡增多,cleaved caspase-3和cytochrome c表达增加(P<0.01);CAT和SOD活性降低,MDA含量和ROS水平增加(P<0.01);SIRT1、NRF2和HO-1表达以及SIRT1活性降低,Ac-FOXO1表达增加(P<0.01);与DOX组相比,DOX+Hes组细胞形态改善,密度和细胞活力增加,培养基中LDH活性降低(P<0.01);细胞凋亡减少,cleaved caspase-3和cytochrome c表达降低(P<0.01);CAT和SOD活性升高,MDA含量和ROS水平降低(P<0.01);SIRT1、NRF2和HO-1表达以及SIRT1活性增加,Ac-FOXO1表达降低(P<0.01);而与DOX+Hes组相比,EX527阻断Hes对DOX诱导的H9c2细胞损伤、氧化应激和SIRT1/NRF2信号的作用。结论Hes可以通过调控SIRT1/NRF2信号抑制氧化应激和细胞凋亡减轻DOX诱导的H9c2心肌细胞毒性。Objective To investigate whether hesperetin(Hes)alleviates doxorubicin(DOX)-induced cardiomyocytotoxicity by reducing oxidative stress via regulating silent information regulator 1(SIRT1)/nuclear transcription factor E2-related factor 2(NRF2)signaling in H9c2 cells.Methods H9c2 cells were treated with DOX to establish the cardiotoxicity model and were randomly assigned to four groups,a control group(Control)and three treatment groups,receiving respectively DOX(the DOX group),Hes+DOX(the DOX+Hes group),and Hes+SIRT1 inhibitor EX527+DOX(the DOX+Hes+EX527 group).Cellular morphology was observed by the light microscope.Cell viability was evaluated by CCK-8.DOX-induced apoptosis in H9c2 cells was examined by flow cytometry.The levels of reactive oxygen species(ROS)in the H9c2 cells of the four groups were determied with 2'-7'-dichlorodihydrofluorescein diacetate(DCFH-DA)staining.The activities of lactate dehydrogenase(LDH),superoxide dismutase(SOD),catalase(CAT),and SIRT1 as well as the malondialdehyde(MDA)content were measured using ELISA kits.The expressions of cleaved caspase-3,cytochrome c,SIRT1,Ac-FOXO1,NRF2,and heme oxygenase 1(HO-1)were determined by Western blot.Results Compared with the Control group,the DOX group showed swollen cellular morphology,decreased cell density and viability,and increased LDH activity in the medium(P<0.01);both apoptosis and the expression of cleaved caspase-3 and cytochrome c increased(P<0.01);the activities of CAT and SOD decreased while the contents of MDA and ROS increased(P<0.01);the expression of SIRT1,NRF2,and HO-1 decreased,the activity of SIRT1 decreased,and the expression of Ac-FOXO1 increased(P<0.01).Compared with the DOX group,the DOX+Hes group showed improved cellular morphology,increased cell density and viability,and decreased LDH activity in the medium(P<0.01);the apoptosis and the expression of cleaved caspase-3 and cytochrome c decreased(P<0.01);the activities of CAT and SOD increased while the levels of MDA and ROS decreased(P<0.01);the expression of SIRT1,NRF2,an

关 键 词：橙皮素 氧化应激 SIRT1/NRF2 H9C2细胞 阿霉素 

分 类 号：R285[医药卫生—中药学]