Background: Gremlin, a bone morphogenetic protein antagonist, plays an important role in the pathogenesis of\r\ndiabetic nephropathy (DN). However, the specific molecular mechanism underlying Gremlinâ��s involvement in DN\r\nhas not been fully elucidated. In the present study, we investigated the role of Gremlin on cell proliferation and\r\naccumulation of extracellular matrix (ECM) in mouse mesangial cells (MMCs), and explored the relationship between\r\nGremlin and the ERK1/2 pathway.\r\nMethods: To determine expression of Gremlin in MMCs after high glucose (HG) exposure, Gremlin mRNA and\r\nprotein expression were evaluated using real-time polymerase chain reaction and western blot analysis, respectively.\r\nTo determine the role of Gremlin on cell proliferation and accumulation of ECM, western blot analysis was used to\r\nassess expression of pERK1/2, transforming growth factor-�Ÿ1 (TGF-�Ÿ1) and connective tissue growth factor (CTGF).\r\nCell proliferation was examined by bromodeoxyuridine (BrdU) ELISA, and accumulation of collagen IV was\r\nmeasured using a radioimmunoassay. This enabled the relationship between Gremlin and ERK1/2 pathway\r\nactivation to be investigated.\r\nResults: HG exposure induced expression of Gremlin, which peaked 12 h after HG exposure. HG exposure alone or\r\ntransfection of normal-glucose (NG) exposed MMCs with Gremlin plasmid (NG + P) increased cell proliferation.\r\nTransfection with Gremlin plasmid into MMCs previously exposed to HG (HG + P) significantly increased this HGinduced\r\nphenomenon. HG and NG + P conditions up-regulated protein levels of TGF-�Ÿ1, CTGF and collagen IV\r\naccumulation, while HG + P significantly increased levels of these further. Inhibition of Gremlin with Gremlin siRNA\r\nplasmid reversed the HG-induced phenomena. These data indicate that Gremlin can induce cell proliferation and\r\naccumulation of ECM in MMCs. HG also induced the activation of the ERK1/2 pathway, which peaked 24 h after HG\r\nexposure. HG and NG + P conditions induced overexpression of pERK1/2, whilst HG + P significantly induced levels\r\nfurther. Inhibition of Gremlin by Gremlin siRNA plasmid reversed the HG-induced phenomena. This indicates\r\nGremlin can induce activation of the ERK1/2 pathway in MMCs.\r\nConclusion: Culture of MMCs in the presence of HG up-regulates expression of Gremlin. Gremlin induces cell\r\nproliferation and accumulation of ECM in MMCs. and enhances activation of the ERK1/2 pathway.
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