title: Knockdown of NYGGF4 (PID1) rescues insulin resistance and mitochondrial dysfunction induced by FCCP in 3T3-L1 adipocytes.
abstract: NYGGF4 is a recently identified gene that is involved in obesity-associated insulin resistance. Previous data from this laboratory have demonstrated that NYGGF4 overexpression might contribute to the development of insulin resistance (IR) and to mitochondrial dysfunction. Additionally, NYGGF4 knockdown enhanced insulin sensitivity and mitochondrial function in 3T3-L1 adipocytes. We designed this study to determine whether silencing of NYGGF4 in 3T3-L1 adipocytes could rescue the effect of insulin sensitivity and mitochondrial function induced by the cyanide p-trifluoromethoxyphenyl-hydrazone (FCCP), a mitochondrion uncoupler, to ascertain further the mechanism of NYGGF4 involvement in obesity-associated insulin resistance. We found that 3T3-L1 adipocytes, incubated with 5μM FCCP for 12h, had decreased levels of insulin-stimulated glucose uptake and had impaired insulin-stimulated GLUT4 translocation. Silencing also diminished insulin-stimulated tyrosinephosphorylation of IRS-1 and serine phosphorylation of Akt. This phenomenon contrasts with the effect of NYGGF4 knockdown on insulin sensitivity and describes the regulatory function of NYGGF4 in adipocytes insulin sensitivity. We next analyzed the mitochondrial function in NYGGF4-silenced adipocytes incubated with FCCP. NYGGF4 knockdown partly rescued the dissipation of mitochondrial mass, mitochondrial DNA, intracellular ATP synthesis, and intracellular reactive oxygen species (ROS) production occurred following the addition of FCCP, as well as inhibition of mitochondrial transmembrane potential (ΔΨm) in 3T3-L1 adipocytes incubated with FCCP. Collectively, our results suggested that addition of silencing NYGGF4 partly rescued the effect of insulin resistance and mitochondrial dysfunction in NYGGF4 silenced 3T3-L1 adipocytes incubated with FCCP, which might explain the involvement of NYGGF4-induced IR and the development of NYGGF4 in mitochondrial function.
authors.affiliation: Department of Pediatrics, Nanjing Maternity and Child Health Hospital of Nanjing Medical University, Nanjing 210004, China.
authors.forename: Chun-Mei
authors.initials: CM
authors.lastname: Shi
authors.forename: Yu-Mei
authors.initials: YM
authors.lastname: Wang
authors.forename: Chun-Mei
authors.initials: CM
authors.lastname: Zhang
authors.forename: Jie
authors.initials: J
authors.lastname: Qiu
authors.forename: Ya-Hui
authors.initials: YH
authors.lastname: Shen
authors.forename: Jin-Gai
authors.initials: JG
authors.lastname: Zhu
authors.forename: Lin
authors.initials: L
authors.lastname: Chen
authors.forename: Guang-Feng
authors.initials: GF
authors.lastname: Xu
authors.forename: Ya-Ping
authors.initials: YP
authors.lastname: Zhao
authors.forename: Chen-Bo
authors.initials: CB
authors.lastname: Ji
authors.forename: Xi-Rong
authors.initials: XR
authors.lastname: Guo
mesh_terms: D017667:Adipocytes / Q000187:drug effects* / Q000378:metabolism*; D000818:Animals; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone / Q000633:toxicity*; D002352:Carrier Proteins / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D055785:Gene Knockdown Techniques; D007333:Insulin Resistance; D051379:Mice; D008928:Mitochondria / Q000378:metabolism
chemical_list: D002352:Carrier Proteins; C556403:NYGGF4 protein, mouse; D002259:Carbonyl Cyanide p-Trifluoromethoxyphenylhydrazone