菊苣酸通过介导Nrf2和NF-κB通路缓解低氧诱导的H9c2心肌细胞氧化应激  

Chicoric acid alleviates oxidative stress via mediating Nrf2 and NF-κB pathways in hypoxia-induced H9c2 cardiomyocytes

作　　者：王文胜 吴华[1] 陈鑫磊 李金铭 陈自鑫 刘嘉逸 许恩榕 陈林 郑润韬 WANG Wensheng;WU Hua;CHEN Xinlei;LI Jinming;CHEN Zixin;LIU Jiayi;XU Enrong;CHEN Lin;ZHENG Runtao(College of Agriculture and Animal Husbandry,Qinghai University,Xining 810016,China)

机构地区：[1]青海大学农牧学院,青海西宁810016

出　　处：《中国兽医科学》2024年第11期1546-1553,共8页Chinese Veterinary Science

基　　金：国家自然科学基金项目(32160812)。

摘　　要：为探讨菊苣酸(CA)通过介导Nrf2和NF-κB通路缓解低氧诱导的H9c2心肌细胞氧化应激的影响,通过构建H9c2心肌细胞体外低氧模型,利用qRT-PCR、Western-blot检测常氧、低氧环境下菊苣酸处理H9c2心肌细胞后Nrf2和NF-κB通路因子的基因转录水平和蛋白表达。结果显示:低氧环境下,Nrf2信号通路关键因子KRAB结合蛋白1(KAP1)基因转录水平极显著降低(P<0.01),Nrf2、血红素氧化酶-1(HO-1)基因转录水平显著降低(P<0.05),KAP1、Nrf2、HO-1、NAD(P)H醌脱氢酶1(NQO1)蛋白表达量均极显著降低(P<0.01);NF-κB信号通路关键因子IκB激酶α(IKKα)、IκB激酶β(IKKβ)、NF-κB、肿瘤坏死因子-α(TNF-α)基因转录水平均极显著升高(P<0.01),IKKα、IKKβ、TNF-α蛋白表达量均极显著升高(P<0.01),NF-κB、环氧合酶-2(COX-2)蛋白表达量显著升高(P<0.05)。添加CA后,KAP1、Nrf2、HO-1、NQO1基因转录水平极显著升高(P<0.01),KAP1、Nrf2蛋白表达量极显著升高(P<0.01);IKKα、IKKβ、NF-κB、TNF-α、COX-2基因转录水平及蛋白表达量均极显著降低(P<0.01)。结果表明,CA可上调H9c2心肌细胞Nrf2通路,抑制NF-κB通路因子的表达,缓解低氧胁迫的氧化应激反应,从而发挥低氧保护作用,以期为开发菊苣酸功能性饲料添加剂提供理论依据。Hypoxia-induced oxidative stress is closely associated with impaired cardiac function.To investigate the effect of chicoric acid(CA)to alleviate oxidative stress via mediating Nrf2 and NF-κB pathways in hypoxia-induced H9c2 cardiomyocytes,a H9c2 cardiomyocyte hypoxia model in vitro was constructed,and utilized q RT-PCR and Western-blot to detect the gene transcriptional levels and protein expression of Nrf2 and NF-κB pathway factors in normoxia,hypoxia,and chicoric acid-treated H9c2cardiomyocytes.The results showed that,under hypoxia environment,Nrf2 signaling pathway key factor KAP1 gene transcriptional level was highly significantly reduced(P<0.01),Nrf2 and HO-1 gene transcriptional level was significantly reduced(P<0.05),and KAP1,Nrf2,HO-1 and NQO1 protein expression were all highly significantly reduced(P<0.01);NF-κB signaling pathway-associated factors IKKα,IKKβ,NF-κB,and TNF-αgene transcriptional levels were all highly significantly elevated(P<0.01),IKKα,IKKβ,and TNF-αprotein expression were all highly significantly elevated(P<0.01),and NF-κB,and COX-2protein expression were significantly elevated(P<0.05).After adding CA,the transcriptional levels of KAP1,Nrf2,HO-1,and NQO1 genes were highly significantly elevated(P<0.01),and the protein expressions of KAP1and Nrf2 were highly significantly elevated(P<0.01);the transcriptional levels and protein expressions of IKKα,IKKβ,NF-κB,TNF-α,and COX-2 genes were highly significantly reduced(P<0.01).The results showed that CA could up-regulate the Nrf2 pathway and inhibit the expression of NF-κB pathway factors in H9c2 cardiomyocytes to alleviate the oxidative stress response in hypoxia stress,thus exerting a hypoxia-protective effect,as to provide a theoretical basis for the development of functional feed additives of chicoric acid.

关 键 词：菊苣酸 低氧 H9C2心肌细胞 氧化应激 Nrf2通路 NF-ΚB通路 

分 类 号：S853.72[农业科学—临床兽医学]