RIP140与TNF-α对心肌细胞能量代谢的调控作用

RIP140 and TNF-α regulate energy metabolism in cardiomyocytes

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 H9c2 cardiomyocytes were infected with Ad-RIP140, simultaneously with or without TNF-α treatment. The mRNA levels of PPAR-α, PPAR-β/δ, and PDK4 were measured. H9c2 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. H9c2 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 H9c2 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.