右美托咪定通过SIRT3去乙酰化TFAM对HK-2细胞缺血再灌注损伤的影响  

Effect of Dexmedetomidine Deacetylated TFAM by SIRT3 on Ischemia-reperfusion Injury of HK-2 Cells

作　　者：胡晨 刘玉丽 HU Chen;LIU Yuli(Huizhou Central People’s Hospital,Huizhou City,Guangdong Province 516001)

机构地区：[1]广东省惠州市中心人民医院,516001

出　　处：《医学理论与实践》2024年第1期12-15,共4页The Journal of Medical Theory and Practice

基　　金：惠州市科技计划项目(221014146940890)。

摘　　要：目的:探讨右美托咪定(Dex)通过沉默信息调节因子2相关酶3(SIRT3)去乙酰化线粒体转录因子A(TFAM)对肾小管上皮细胞(HK-2)缺血再灌注损伤(IRI)的影响。方法:人HK-2细胞株分为对照组、缺血再灌注(I/R)组、Dex组、SIRT3抑制剂(3-TYP)组及Dex+3-TYP组。除对照组外,其余各组均制备I/R模型,Dex组、3-TYP组及Dex+3-TYP组分别在I/R制备前分别给予Dex、3-TYP及Dex+3-TYP处理;I/R组及对照组在建模前加入等量生理盐水处理。观察各组细胞培养24h、48h的活性,检测细胞炎症因子[白介素细胞6(IL-6)、IL-8、IL-10]水平。提取线粒体,检测活性氧(ROS)、线粒体膜电位(MMP)、线粒体通透性转换孔(mPTP)开放程度及线粒体DNA(mtDNA)数量。免疫共沉淀(Co-IP)验证SIRT3与TFAM是否相互作用。结果:I/R模型建立后,IL-6、IL-8、ROS水平及TFAM乙酰化水平均升高,细胞活力(24/48h的OD值)、IL-10、MMP、mPTP、mtDNA水平及SIRT3表达均下降(P<0.05);I/R模型经3-TYP干预后上述变化加重(P<0.05);Dex干预I/R模型后,细胞活力增加,IL-10、MMP、mPTP、mtDNA水平及SIRT3表达均升高,IL-6、IL-8、ROS水平及TFAM乙酰化水平均下降(P<0.05);3-TYP与Dex共干预后Dex作用减弱(P<0.05);Co-IP验证结果显示SIRT3与TFAM均被沉淀,二者存在相互作用。结论:SIRT3去乙酰化TFAM参与Dex减轻HK-2细胞缺血再灌注损伤的过程。Objective:To investigate the effects of dexmedetomidine(Dex)on ischemia-reperfusion injury(IRI)of renal tubular epithelial cells(HK-2)by silence information regulator 2 homolog 3(SIRT3)to deacetylate mitochondrial transcription factor A(TFAM).Methods:HK-2 cell lines were divided into control group,ischemia-reperfusion(I/R)group,Dex group,SIRT3 inhibitor(3-TYP)group and Dex+3-TYP group.I/R models were prepared in all groups except the control group.Dex group,3-TYP group and Dex+3-TYP group were treated with Dex,3-TYP and Dex+3-TYP,respectively,before I/R preparation.I/R group and control group were treated with equal amount of normal saline before modeling.The activity of cells cultured for 24 hours and 48 hours in each group was observed.The levels of cellular inflammatory cytokines[interleukin ccell 6(IL-6),IL-8,IL-10]were detected.Mitochondria were extracted to detect reactive oxygen species(ROS),mitochondrial membrane potential(MMP),mitochondrial permeability transition pore(mPTP)openness and mitochondrial DNA(mtDNA)quantity.Co-immunoprecipitation(Co-IP)was used to verify the interaction between SIRT3 and TFAM.Results:After the establishment of I/R model,the levels of IL-6,IL-8,ROS and TFAM were increased,while the cell viability(OD value 24/48 hours),IL-10,MMP,mPTP,mtDNA and SIRT3 expression were decreased(P<0.05).The above changes were aggravated in I/R model after 3-TYP intervention(P<0.05).After Dex intervention in I/R model,cell viability was increased,IL-10,MMP,mPTP,mtDNA levels and SIRT3 expression were increased,IL-6,IL-8,ROS levels and TFAM acetylation levels were decreased(P<0.05),Dex effect was weakened after 3-TYP and Dex intervention(P<0.05).Co-IP verification results showed that both SIRT3 and TFAM were precipitated,and there was interaction between the two.Conclusion:SIRT3 deacetylation of TFAM is involved in the reduction of ischemia-reperfusion injury of HK-2 cells by Dex.

关 键 词：肾小管上皮细胞 缺血再灌注损伤 右美托咪定 沉默信息调节因子2相关酶3 线粒体转录因子A 

分 类 号：R36[医药卫生—病理学] R692[医药卫生—基础医学]