FOXM1通过靶向上调PHB2的转录改善线粒体功能障碍保护脂多糖诱导的肾小管上皮细胞损伤

doi:10.3969/j.issn.1006-6233.2022.03.04

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摘要 目的:探究FOXM1在脂多糖(LPS)诱导的肾小管上皮细胞损伤中的作用及其可能的作用机制。方法:体外培养人肾小管上皮细胞HK-2,qRT-PCR和Western blot检测细胞FOXM1和PHB2表达;CCK-8检测细胞活力;流式细胞术检测细胞凋亡;JC-1染色检测线粒体膜电位;试剂盒检测细胞ATP含量;荧光素酶报告实验和ChIP-PCR实验检测FOXM1对PHB2启动子调控作用。结果:与对照组相比,LPS组细胞中FOXM1和PHB2表达、细胞活力、红/绿荧光强度比值和ATP含量明显降低(P<0.05),细胞凋亡率明显升高(P<0.05);与LPS组比较,LPS+oe-NC组细胞中FOXM1和PHB2表达、细胞活力、红/绿荧光强度比值、ATP含量和细胞凋亡率差异无统计学意义(P>0.05);与LPS+oe-NC组相比,LPS+oe-FOXM1组细胞中FOXM1和PHB2表达、细胞活力、红/绿荧光强度比值和ATP含量明显升高(P<0.05),细胞凋亡率明显降低(P<0.05)。FOXM1靶向结合PHB2启动子区域。与LPS+oe-NC+ sh-NC组相比,LPS+oe-FOXM1+sh-NC组细胞中FOXM1和PHB2蛋白表达、细胞活力、红/绿荧光强度比值和ATP含量明显升高(P<0.05),细胞凋亡率明显降低(P<0.05);与LPS+oe-NC+sh-NC组相比,LPS+oe-NC+sh-PHB2组细胞中PHB2蛋白表达、细胞活力、红/绿荧光强度比值和ATP含量明显降低(P<0.05),细胞凋亡率明显升高(P<0.05),FOXM1蛋白表达差异无统计学意义(P>0.05);sh-PHB2可部分逆转oe-FOXM1对LPS处理的HK-2细胞的作用。结论:FOXM1通过靶向结合PHB2启动子区域促进PHB2表达,改善线粒体功能障碍,从而抑制LPS诱导的肾小管上皮细胞损伤。

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关键词 ：肾小管上皮细胞, 脂多糖, Forkhead box M1, prohibitin 2, 线粒体

Abstract：Objective: To explore the role of FOXM1 in lipopolysaccharide (LPS)-induced renal tubular epithelial cell injury and its possible mechanism. Methods: Human renal tubular epithelial cells HK-2 were cultured in vitro. QRT-PCR and Western blot were used to detect the expression of FOXM1 and PHB2; CCK-8 was used to detect cell viability; flow cytometry was used to detect cell apoptosis; JC-1 staining was used to detect the mitochondrial membrane potential; the reagent kit was used to detect cellular ATP content; the luciferase reporter experiment and ChIP-PCR experiment were used to detect the regulatory effect of FOXM1 on the PHB2 promoter. Results: Compared with the control group, the expression of FOXM1 and PHB2, cell viability, red/green fluorescence intensity ratio and ATP content in the cells of the LPS group were significantly reduced (P<0.05), and the apoptosis rate was significantly increased (P<0.05); compared with the LPS group, there was no significant difference in FOXM1 and PHB2 expression, cell viability, red/green fluorescence intensity ratio, ATP content and apoptosis rate in the cells of the LPS+oe-NC group (P>0.05); compared with the LPS+oe-NC group, the expression of FOXM1 and PHB2, cell viability, red/green fluorescence intensity ratio and ATP content in the cells of the LPS+oe-FOXM1 group were significantly increased (P<0.05), and the apoptosis rate was significantly reduced (P<0.05). FOXM1 targeted the PHB2 promoter region. Compared with the LPS+oe-NC+sh-NC group, the FOXM1 and PHB2 protein expression, cell viability, red/green fluorescence intensity ratio and ATP content in the cells of the LPS+oe-FOXM1+sh-NC group were significantly increased (P<0.05), the cell apoptosis rate was significantly reduced (P＜0.05); compared with the LPS+oe-NC+sh-NC group, the expression of PHB2 protein, cell viability, red/green fluorescence intensity ratio and ATP content in the cells of the LPS+oe-NC+sh-PHB2 group were significantly reduced (P＜0.05), the apoptosis rate was significantly increased (P<0.05), and the difference in FOXM1 protein expression was not statistically significant (P>0.05); sh-PHB2 can partially reverse the effect of oe-FOXM1 on HK-2 cells treated with LPS. Conclusion: FOXM1 promotes PHB2 expression by targeting the PHB2 promoter region to improve mitochondrial dysfunction, and then to inhibit LPS-induced renal tubular epithelial cell damage.

Key words： Renal tubular epithelial cells Lipopolysaccharide Forkhead box M1 Prohibitin 2 Mitochondria

基金资助:国家自然科学基金项目,(编号:81560124);海南省卫生健康行业科研项目,(编号:20A200165)

通讯作者: 王善志

引用本文:

陈娟, 陈拉斯, 陈丽, 唐文庄, 张琼果, 王善志. FOXM1通过靶向上调PHB2的转录改善线粒体功能障碍保护脂多糖诱导的肾小管上皮细胞损伤[J]. 河北医学, 2022, 28(3): 368-374.
CHEN Juan, CHEN Lasi, CHEN Li, et al. FOXM1 Ameliorates Mitochondrial Dysfunction by Targeting Upregulation of PHB2 Transcription to Protect Against Lipopolysaccharide-Induced Renal Tubular Epithelial Cell Injury. HeBei Med, 2022, 28(3): 368-374.

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http://www.hbyxzzs.cn/CN/10.3969/j.issn.1006-6233.2022.03.04 或 http://www.hbyxzzs.cn/CN/Y2022/V28/I3/368

[1] Ronco C,Bellomo R,Kellum JA.Acute kidney injury[J].Lancet,2019,394(10212):1949~1964.
[2] Sweeney RP,Lowary TL.New insights into lipopolysaccharide assembly and export[J].Curr Opin Chem Biol,2019,53(4):37~43.
[3] Liu X,Lu J,Liao Y,et al.Dihydroartemisinin attenuates lipopolysaccharide-induced acute kidney injury by inhibiting inflammation and oxidative stress[J].Biomed Pharmacother,2019,117(2):1090~1098.
[4] O'Regan RM,Nahta R.Targeting forkhead box M1 transcription factor in breast cancer[J].Biochem Pharmacol,2018,154(7):407~413.
[5] Zhang FM,Zheng WH,Wang HJ.MiR-34a-5p inhibition attenuates LPS-induced endothelial cell injury by targeting FOXM1[J].Eur Rev Med Pharmacol Sci,2020,24(20):10829~10838.
[6] Signorile A,Sgaramella G,Bellomo F,et al.Prohibitins:a critical role in mitochondrial functions and implication in diseases[J].Cells,2019,8(1):71~79.
[7] Xu Y,Wang J,Xu W,et al.Prohibitin 2-mediated mitophagy attenuates renal tubular epithelial cells injury by regulating mitochondrial dysfunction and NLRP3 inflammasome activation[J].Am Physiol Renal Physiol,2019,316(2):F396~F407.
[8] Prowle JR.Sepsis-associated AKI[J].Clin Am Soc Nephrol,2018,13(2):339~342.
[9] Chan DC.Mitochondrial dynamics and its involvement in disease[J].Annu Rev Pathol,2020,15(5):235~259.
[10] Annesley SJ,Fisher PR.Mitochondria in health and disease[J].Cells,2019,8(7):680~689.
[11] Sun J,Zhang J,Tian J,et al.Mitochondria in sepsis-induced AKI[J].Am Soc Nephrol,2019,30(7):1151~1161.
[12] Xiang X,Fu Y,Zhao K,et al.Cellular senescence in hepatocellular carcinoma induced by a long non-coding RNA-encoded peptide PINT87aa by blocking FOXM1-mediated PHB2[J].Theranostics,2021,11(10):4929~4944.
[13] Chang-Panesso M,Kadyrov FF,Lalli M,et al.FOXM1 drives proximal tubule proliferation during repair from acute ischemic kidney injury[J].Clin Invest,2019,129(12):5501~5517.
[14] Sinha S,Dwivedi N,Woodgett J,et al.Glycogen synthase kinase-3β inhibits tubular regeneration in acute kidney injury by a FoxM1-dependent mechanism[J].FASEB,2020,34(10):13597~13608.
[15] Wang D,Tabti R,Elderwish S,et al.Prohibitin ligands:a growing armamentarium to tackle cancers,osteoporosis,inflammatory,cardiac and neurological diseases[J].Cell Mol Life Sci,2020,77(18):3525~3546.
[16] Wang J,Zhu P,Li R,et al.Bax inhibitor 1 preserves mitochondrial homeostasis in acute kidney injury through promoting mitochondrial retention of PHB2[J].Theranostics,2020,10(1):384~397.