NYGGF4is a recently identified gene that is involved in obesity-associated insulin resistance.Previous data from this laboratory has been demonstrated that NYGGF4 overexpression might be contribute to the development of insulin resistance and mitochondrial dysfunction;additionally,NYGGF4knockdown enhanced insulin sensitivity and mitochondrial function of 3T3-L1 adipocytes.In this study,we designed to define whether silencing NYGGF4 in 3T3-L1 adipocytes could resuce the effect of insulin sensitivity and mitochondrial function induced by the cyanide p-trifluoromethoxyphenyl-hydrazone(FCCP),a mitochondrial uncoupler,in order to further ascertain the mechanism of NYGGF4 involved in obesity-associated insulin resistance.We found that NYGGF4 silenced adipocytes were incubated with 5 μM and 7.5 μM FCCP for 12h,resulting in decreased basal and insulin-stimulated glucose uptake and impaired insulin-stimulated GLUT4translocation.It also diminished insulin-stimulated tyrosinephosphorylation of IRS-1 and serine phosphorylation of Akt,which are converse with the effect of NYGGF4knockdownon insulin sensitivity and ascertained the regulatory function of NYGGF4 in adipocytes insulin sensitivity.We next analyzed the mitochondrial function in NYGGF4 silenced adipocytes incubated with FCCP.Dissipation of mitochondrial mass,mitochondrial DNA,intracellular ATP synthesis,and intracellular ROS production with FCCP,as well as inhibition of mitochondrial transmembrane potential(△Ψm)in NYGGF4silenced adipocytes.Collectively,our results suggested that FCCP was fully reverse the effect of insulin sensitivity and mitochondrial function in NYGGF4 silenced 3T3-L1 adipocytes,which might be responsible for the role of NYGGF4-induced insulin resistance and the development of NYGGF4 in mitochondrial function.
Chun-mei SHI Chen-bo JI Xi-rong GUO Yu-mei WANG Chun-mei ZHANG Jie QIU Ya-hui SHEN Jin-gai ZHU Lin-CHEN Guang-feng XU Ya-ping ZHAO
Department of Pediatrics,Nanjing Maternity and Child Health Hospital of Nanjing Medical University,N Department of Pediatrics,Nanjing Maternity and Child Health Hospital of Nanjing Medical University,N Department of Pediatrics,Nanjing Maternity and Child Health Hospital of Nanjing Medical University,N Department of Child Health,Huai” an Maternity and Child Health Hospital,Huai” an 223002,China Department of Pediatrics,Nanjing Maternity and Child Health Hospital of Nanjing Medical University,N Institute of Pediatrics of Nanjing Medical University,Nanjing 210029,China