NSUN2对阿霉素诱导H9C2细胞损伤的作用及机制  

Effect and Mechanism of NSUN2 on Doxorubicin-induced Damage in H9C2 Cells

作　　者：郑丽娜[1] 王洁[1] 程飞[1] 陈佳娟[1] 丁妍[1] 王治校[1] ZHENG Li-na;WANG Jie;CHENG Fei;CHEN Jia-juan;DING Yan;WANG Zhi-xiao(Department of Cardiology,Taihe Hospital,Hubei Universi-ty of Medicine,Shiyan,Hubei 442000,China)

机构地区：[1]湖北医药学院附属太和医院心内科,湖北十堰442000

出　　处：《湖北医药学院学报》2023年第3期252-257,F0002,共7页Journal of Hubei University of Medicine

基　　金：国家自然科学基金面上项目(82270530);湖北省自然科学基金创新群体项目(2022CFA036)。

摘　　要：目的:探讨NSUN2对阿霉素(doxorubicin, DOX)诱导H9C2细胞损伤的作用及可能机制。方法:H9C2细胞分两组,对照组(加入0.9%氯化钠溶液2μL)和DOX组加入用0.9%氯化钠溶液配置成0.5 mg/mL阿霉素2μL、siNSUN2转染H9C2细胞、腺病毒转染人源NSUN2建立NSUN2稳定过表达H9C2细胞系,加入阿霉素500 ng/mL共培养24 h后,用DCFH-DA探针、Western blot、免疫荧光、TUNEL染色等方法检测细胞的活性氧(ROS)、凋亡及NSUN2的变化,ELISA检测细胞培养基上清内NSUN2表达变化。结果:Western blot结果显示,DOX组与对照组比较,H9C2细胞Bax蛋白水平及NSUN2表达水平升高(P<0.01),Bcl-2蛋白水平下降(P<0.01);ELISA检测细胞培养液内NSUN2表达水平DOX组较对照组升高(P<0.05);TUNEL结果显示DOX组细胞凋亡明显增加;DCFH-DA探针检测DOX组细胞内ROS水平较对照组明显升高;H9C2细胞敲减NSUN2后,DOX组细胞内Bax蛋白水平较对照组升高(P<0.01),Bcl-2蛋白表达水平下降(P<0.01);NSUN2过表达时,DOX组与对照组比较,细胞内Bax表达水平下降(P<0.01),Bcl-2表达水平升高(P<0.01)。结论:NSUN2能抑制阿霉素诱导的H9C2细胞损伤,对其H9C2细胞起保护作用,机制可能与NSUN2抗氧化应激相关。Objective To explore the effect and possible mechanism of NSUN2 on doxorubicin(DOX)-induced H9C2 cell injury.Methods H9C2 cells were divided into two groups,control group(add 2μL of 0.9%sodium chloride solution)and DOX group(add 0.5 mg/mL doxorubicin).H9C2 cells were transfected with siNSUN2 and human NSUN2 to establish sta⁃ble NSUN2 overexpression H9C2 cell lines.After 500 ng/mL doxorubicin was added for co-culture for 24 hours,the chan⁃ges of ROS,apoptosis and NSUN2 were detected by DCFH-DA probe,Western blot,immunofluorescence and TUNEL staining,and the changes of NSUN2 expression in cell culture supernatant were detected by ELISA.Results Western blot results showed that compared with the control group,Bax protein level and NSUN2 expression level of H9C2 cells in DOX group were increased(P<0.01),while Bcl-2 protein level was decreased(P<0.01).The expression level of NSUN2 in cell culture medium detected by ELISA was higher in the DOX group than that in the control group(P<0.05).The TUNEL results showed a significant increase in cell apoptosis in the DOX group.The intracellular ROS levels in the DOX group were significantly higher than those in the control group detected by the DCFH-DA probe.After knocking down NSUN2 in H9C2 cells,the intracellular Bax protein level in the DOX group increased compared to the control group(P<001),while the Bcl-2 protein expression level decreased(P<0.01).When NSUN2 was overexpressed,compared with the control group,the DOX group showed a decrease in intracellular Bax expression level(P<0.01)and an increase in Bcl-2 expres⁃sion level(P<0.01).Conclusion NSUN2 can inhibit doxorubicin-induced H9C2 cell damage and has protective effect on H9C2 cells.The mechanism may be related to the anti-oxidative stress of NSUN2.

关 键 词：NSUN2 阿霉素 H9C2细胞 活性氧 

分 类 号：R965[医药卫生—药理学]