海藻多糖调节Nrf2通路对过氧化氢诱导的心肌细胞损伤及铁死亡的抑制作用  

Inhibitory effect of seaweed polysaccharide on cardiomyocyte injury and ferroptosis induced by hydroperoxide through regulating pathway of nuclear factor E2 related factor 2

作　　者：张联标 杨荧 王东升 Zhang Lianbiao;Yang Ying;Wang Dongsheng(Department of Cardiovascular Medicine,419th Hospital of Nuclear Industry,Shaoguan,512026,China;不详)

机构地区：[1]核工业四一九医院心血管内科,韶关512026 [2]韶关学院医学院临床系,韶关512026

出　　处：《中国循证心血管医学杂志》2023年第6期712-715,720,共5页Chinese Journal of Evidence-Based Cardiovascular Medicine

摘　　要：目的研究海藻多糖调节Nrf2通路对过氧化氢(H_(2)O_(2))诱导的H9C2心肌细胞损伤及铁死亡的抑制作用。方法采用H_(2)O_(2)诱导法建立心肌细胞损伤模型,设置正常对照组、氧化损伤组、海藻多糖低、中、高剂量组及阳性对照组,海藻多糖各剂量组分别加入0.3、0.6、1.2 mg/ml的海藻多糖,阳性对照组加入50μmol/L的叔丁基对苯二酚(tBHQ),继续培养72 h,使用噻唑蓝比色(MTT)法测定H9C2细胞存活率,酶联免疫吸附法(ELISA)测定H9C2细胞乳酸脱氢酶(LDH)漏出率,H9C2细胞Fe^(2+)、活性氧(ROS)、丙二醛(MDA)、4-羟基壬烯醛(4-HNE)、还原型谷胱甘肽(GSH)水平及H9C2细胞上清液LDH、肌酸激酶(CK)水平,流式细胞法测定H9C2细胞凋亡率,实时定量聚合酶链式反应(RT-qPCR)测定H9C2细胞Nrf2、GPX4及HO-1 mRNA水平,免疫印迹法(Western blot)测定H9C2细胞Nrf2、GPX4及HO-1蛋白水平。结果与氧化损伤组比较,海藻多糖各剂量组及阳性对照组H9C2细胞存活率、GSH水平,H9C2细胞Nrf2、GPX4及HO-1 mRNA及蛋白水平明显升高(P<0.05),H9C2细胞Fe^(2+)、ROS、MDA、4-HNE水平、LDH漏出率、上清液LDH及CK水平,H9C2细胞凋亡率明显降低(P<0.05)。结论海藻多糖能够抑制过氧化氢诱导的H9C2心肌细胞氧化损伤,改善H9C2铁死亡,抑制H9C2心肌细胞死亡,其机制可能与调节Nrf2通路有关。Objective To study the inhibitory effect of seaweed polysaccharide(SPS)on injury of H9C2 cardiomyocytes and ferroptosis induced by hydroperoxide(HP)through regulating pathway of nuclear factor E2 related factor 2(Nrf2).Methods The model of cardiomyocyte injury was established by using HP induction method,and then normal control group,oxidative damage group,low-dose SPS group(given 0.3 mg/ml of SPS),mid-dose SPS group(given 0.6 mg/ml of SPS),high-dose SPS group(given 1.2 mg/ml of SPS)and positive control group(given 50μmol/L of tBHQ).The survival rate of H9C2 cardiomyocytes was detected by using MTT method,and leakage rate of lactic dehydrogenase(LDH),levels of Fe^(2+),ROS,MDA,4-HNE and GSH in H9C2 cardiomyocytes and levels of LDH and CK in supernatant of H9C2 cardiomyocytes were detected by using ELISA.The apoptosis rate of H9C2 cardiomyocytes was detected by using flow cytometry(FCM),and levels of Nrf2,GPX4 and HO-1 mRNA in H9C2 cardiomyocytes were detected by using RT-qPCR.The levels of Nrf2,GPX4 and HO-1 protein in H9C2 cardiomyocytes were detected by using Western blotting assay.Results The survival rate of H9C2 cardiomyocytes,and levels of GSH,Nrf2,GPX4 and HO-1 mRNA and protein in H9C2 cardiomyocytes increased significantly(P<0.05),and levels of Fe^(2+),ROS,MDA and 4-HNE,LDH leakage rate in H9C2 cardiomyocytes,levels of LDH and CK in supernatant,and apoptosis rate of H9C2 cardiomyocytes decreased significantly(P<0.05)in all SPS groups and positive control group compared with oxidative damage group.Conclusion SPS can inhibit oxidative damage of H9C2 cardiomyocytes induced by HP,reduce ferroptosis and death of H9C2 cardiomyocytes,and the mechanism may be related to Nrf2 pathway regulation.

关 键 词：心肌细胞损伤 海藻多糖 铁死亡 Nrf2通路 

分 类 号：R542.2[医药卫生—心血管疾病]