RIP140与TNF-α对心肌细胞能量代谢的调控作用
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摘要
目的观察RIP140与TNF-α对心肌细胞能量代谢的影响。方法体外培养H9c2心肌细胞,分为空载病毒组、过表达RIP140组、空载病毒+TNF-α组、过表达RIP140+TNF-α组,荧光定量PCR检测PPAR-α、PPAR-β/δ、PDK4的mRNA表达水平。使用过表达RIP140的腺病毒感染H9c2细胞,Western blot分析细胞核p65蛋白水平、胞质IκB-α蛋白水平;荧光定量PCR检测TNF-α、IL-1β、IL-2的mRNA表达水平; TNF-α刺激心肌细胞,荧光定量PCR检测RIP140的mRNA表达水平,Western blot分析RIP140蛋白表达水平。结果与过表达RIP140组比较,过表达RIP140+TNF-α刺激组PPAR-β/δ和PDK4的mRNA表达下降。过表达RIP140的心肌细胞核内p65蛋白水平升高,胞质IκB-α蛋白水平下降,TNF-α、IL-1β、IL-2的mRNA表达升高; TNF-α刺激心肌细胞,使RIP140的mRNA和蛋白表达水平升高。结论RIP140与TNF-α可相互作用,介导心肌细胞炎症反应和能量代谢紊乱。
Aim To explore whether RIP140 and TNF-α regulate energy metabolism in cardiomyocytes. Methods H9 c2 cardiomyocytes were infected with AdRIP140,simultaneously with or without TNF-α treatment. The mRNA levels of PPAR-α,PPAR-β/δ,and PDK4 were measured. H9 c2 was exposed to adenovirus expressing RIP140-specific or nonspecific control. Expression of p65 in the nucleus and IκB-α in cytoplasm were measured by Western blotting,and mRNA levels of IL-1β,IL-2 and TNF-α were measured by real-time PCR. H9 c2 was treated with or without TNF-α. The mRNA and protein levels of RIP140 were measured.Results Overexpression of RIP140 led to a decrease in mRNA levels of PPAR-α, PPAR-β/δ, PDK4,while TNF-α aggravated down-regulation of key metabolic genes by superabundant RIP140. A marked increase of p65-NF-κB in nuclear,a significant decrease of IκB-α in cytoplasm and a notable increase in mRNA levels of TNF-α,IL-1β and IL-2 in H9 c2 cell line were observed following overexpression of RIP140. The mRNA and protein levels of RIP140 were up-regulated by TNF-α treatment. Conclusions RIP140 and TNF-α may collaborate in mediating proinflammatory processes and metabolic dysregulation in cardiomyocytes.
Aim To explore whether RIP140 and TNF-α regulate energy metabolism in cardiomyocytes. Methods H9 c2 cardiomyocytes were infected with AdRIP140,simultaneously with or without TNF-α treatment. The mRNA levels of PPAR-α,PPAR-β/δ,and PDK4 were measured. H9 c2 was exposed to adenovirus expressing RIP140-specific or nonspecific control. Expression of p65 in the nucleus and IκB-α in cytoplasm were measured by Western blotting,and mRNA levels of IL-1β,IL-2 and TNF-α were measured by real-time PCR. H9 c2 was treated with or without TNF-α. The mRNA and protein levels of RIP140 were measured.Results Overexpression of RIP140 led to a decrease in mRNA levels of PPAR-α, PPAR-β/δ, PDK4,while TNF-α aggravated down-regulation of key metabolic genes by superabundant RIP140. A marked increase of p65-NF-κB in nuclear,a significant decrease of IκB-α in cytoplasm and a notable increase in mRNA levels of TNF-α,IL-1β and IL-2 in H9 c2 cell line were observed following overexpression of RIP140. The mRNA and protein levels of RIP140 were up-regulated by TNF-α treatment. Conclusions RIP140 and TNF-α may collaborate in mediating proinflammatory processes and metabolic dysregulation in cardiomyocytes.
引文
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[12]Chen Y F,Wang Y H,Chen J W,et al.Roles of transcriptional corepressor RIP140 and coactivator PGC-1 alpha in energy state of chronically infarcted rat hearts and mitochondrial function of cardiomyocytes[J].Mol Cell Endocrinol,2012,362(1-2):11-8.
[2]陈艳芳,张銮坤,王若伦,刘培庆.腺病毒介导心脏特异性过表达RIP140对大鼠心脏功能及炎症通路的影响[J].中国药理学通报,2017,33(8):1068-72.[2]Chen Y F,Zhang L K,Wang R L,Liu P Q.Role of adenovirusmediated cardiac-specific RIP140 overexpression in cardiac function and inflammation pathway[J].Chin Pharmacol Bull,2017,33(8):1068-72.
[3]Schilling J,Lai L,Sambandam N,et al.Toll-like receptor-mediated inflammatory signaling reprograms cardiac energy metabolism by repressing peroxisome proliferator-activated receptor gamma coactivator-1 signaling[J].Circ Heart Fail,2011,4(4):474-82.
[4]Palomer X,Alvarez-Guardia D,Rodriguez-Calvo R,et al.TNF-alpha reduces PGC-1alpha expression through NF-kappaB and p38MAPK leading to increased glucose oxidation in a human cardiac cell model[J].Cardiovasc Res,2009,81(4):703-12.
[5]陈艳芳,王若伦,刘培庆.RIP140重组腺病毒的构建和在乳鼠心肌细胞中的表达[J].中国药理学通报,2016,32(12):1735-40.[5]Chen Y F,Wang R L,Liu P Q.Construction of RIP140 recombinant adenovirus and its expression in neonatal rat cardiomyocytes[J].Chin Pharmacol Bull,2016,32(12):1735-40.
[6]Wong S C,Fukuchi M,Melnyk P,et al.Induction of cyclooxygenase-2 and activation of nuclear factor-kappaB in myocardium of patients with congestive heart failure[J].Circulation,1998,98(2):100-3.
[7]Freund C,Schmidt-Ullrich R,Baurand A,et al.Requirement of nuclear factor-kappa B in angiotensin II-and isoproterenol-induced cardiac hypertrophy in vivo[J].Circulation,2005,111(18):2319-25.
[8]Maier H J,Schips T G,Wietelmann A,et al.Cardiomyocyte-specific I kappa B kinase(IKK)/NF-kappa B activation induces reversible inflammatory cardiomyopathy and heart failure[J].Proc Natl Acad Sci USA,2012,109(29):11794-9.
[9]Chen Y F,Chen S R,Yue Z B,et al.Receptor-interacting protein 140 overexpression impairs cardiac mitochondrial function and accelerates the transition to heart failure in chronically infarcted rats[J].Transl Res,2017,180:91-102.e1.
[10]Ding G,Cheng L,Qin Q,et al.PPARdelta modulates lipopolysaccharide-induced TNFalpha inflammation signaling in cultured cardiomyocytes[J].J Mol Cell Cardiol,2006,40(6):821-8.
[11]Zhang L K,Chen Y F,Yue Z B,et al.The p65 subunit of NF-kappaB involves in RIP140-mediated inflammatory and metabolic dysregulation in cardiomyocytes[J].Arch Biochem Biophys,2014,554:22-7.
[12]Chen Y F,Wang Y H,Chen J W,et al.Roles of transcriptional corepressor RIP140 and coactivator PGC-1 alpha in energy state of chronically infarcted rat hearts and mitochondrial function of cardiomyocytes[J].Mol Cell Endocrinol,2012,362(1-2):11-8.
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