Resumen de Ectopic UCP1 gene expression in HepG2 cells ffects ATP production

Pedro González Muniesa, Javier Campión Zabalza, José Alfredo Martínez Hernández, Fermín Ignacio Milagro Yoldi

• español

  The UCP1 is an uncoupling protein located in the inner mitochondrial membrane of brown adipocytes, which has a well-documented role in diet-induced thermogenesis. The current study assessed whether UCP1 transfected liver cells demand more fuel substrates in the oxidative phosphorylation processes. Therefore, the purpose of this experiment was to achieve an ectopic expression of UCP1 in HepG2 cells to significantly decrease the production of ATP. The UCP1 gene was transferred into the hepatic cells by using a calcium phosphate precipitation protocol. The efficiency of the transfection was tested, 48 hours later, by bioluminescence of luciferase previously transfected, while the expression of mRNA of UCP1 was demonstrated by RT-PCR. In addition, measuring the production of ATP by using a bioluminescence procedure assessed the functionality of this protein. Transfected liver cells with UCP1 showed a decrease of 23% in ATP production in comparison with control cells without expression of UCP1 (2.23 vs. 2.90 RLU/pg protein, p=0.015). In conclusion, the ectopic expression of UCP1 decreased the production of ATP, possibly uncoupling the oxidative phosphorylation, which could be a novel approach for understanding thermogenic processes and eventually for energy metabolism and body weight management.

• English

  The UCP1 is an uncoupling protein located in the inner mitochondrial membrane of brown adipocytes, which has a well-documented role in diet-induced thermogenesis. The current study assessed whether UCP1 transfected liver cells demand more fuel substrates in the oxidative phosphorylation processes. Therefore, the purpose of this experiment was to achieve an ectopic expression of UCP1 in HepG2 cells to significantly decrease the production of ATP. The UCP1 gene was transferred into the hepatic cells by using a calcium phosphate precipitation protocol. The efficiency of the transfection was tested, 48 hours later, by bioluminescence of luciferase previously transfected, while the expression of mRNA of UCP1 was demonstrated by RT-PCR. In addition, measuring the production of ATP by using a bioluminescence procedure assessed the functionality of this protein. Transfected liver cells with UCP1 showed a decrease of 23% in ATP production in comparison with control cells without expression of UCP1 (2.23 vs. 2.90 RLU/pg protein, p=0.015). In conclusion, the ectopic expression of UCP1 decreased the production of ATP, possibly uncoupling the oxidative phosphorylation, which could be a novel approach for understanding thermogenic processes and eventually for energy metabolism and body weight management.