Pannexin1-FAM134B Dependent Reticulophagy Mediates Apelin-13 Induced Cardiomyocytes Hypertrophy

Aim: Apelin is an endogenous ligand for the G protein-coupled receptor APJ.Pannexin1 is a gap junctional protein which involves in the development of cardiovascular related diseases.FAM134 B is receptor of reticulophagy which is the key receptor that mediated the reticulophagy happens.Thus,in our study we try to investigate the effect of apelin-13 onpannexin1-FAM134 B dependent reticulophagy metabolism.Methods: 1.The expression of pannexin1,FAM134 B,Beclin-1,BNP and LC3II/Iin H9c2 cells were tested by Western Blot;2.The expression of apelin,APJ,pannexin1,FAM134 B,Beclin-1 and BNP in human autopsy cardiac hypertrophy heart tissue;3.Design si RNA interfere chains to interfere FAM134 B expression;4.The diameter and volume of H9c2 cells was tested by Scepter TM Handheld cell counter;5.The concentration of proteins in H9c2 cells was tested by BCA protein quantitation kit.Results: 1.Apelin-13 promotes the expression of pannexin1,FAM134 B,Beclin-1 and LC3II/I in a concentration(0-1μmol/L)dependent manner;2.Apelin-13 promotes the expression of pannexin1,FAM134 B,Beclin-1 and LC3II/I in a time(0-24h)dependent manner;3.APJ antagonist F13A(1μmol/L,24h)blocks the effect of apelin-13 on pannexin1,FAM134 B,Beclin-1,LC3II/I expression in H9c2 cells;4.Pannexin1 inhibitors probenecid(200μmol/L,24h)can significant inhibits the apelin-13(1μmol/L,24h)induced the expression of pannexin1,FAM134 B and Beclin-1in H9c2 cells;5.Pannexin1 inhibitors carbenoxolone(10μmol/L,24h)can significant inhibits the apelin-13(1μmol/L,24h)induced the expression of pannexin1,FAM134 B,Beclin-1 and LC3II/I in H9c2 cells;6.Pannexin1 inhibitor probenecid(200μmol/L,96h)blocks apelin-13(1μmol/L,96h)promoted diameter and volume of H9c2 cells and BNP expression;7.Pannexin1 inhibitor carbenoxolone(10μmol/L,96h)blocks apelin-13(1μmol/L,96h)promoted diameter and volume of H9c2 cells and BNP expression;8.si Fam134 B suppresses apelin-13(1μmol/L,96h)promoted diameter and volume of H9c2 cells.9.The protein of apelin,APJ,pannexin1,FAM134 B and Beclin-1 are high expression in human autopsy cardiac hypertrophy heart tissue;Conclusion : Pannexin1-FAM134 B dependent Reticulophagy mediates apelin-13 induced cardiomyocytes hypertrophy...