dopamine receptors in human adipocytes expression and functions多巴胺受体在人类脂肪细胞表达和功能.pdfVIP

Dopamine Receptors in Human Adipocytes: Expression and Functions 1 1 1 1 1 Dana C. Borcherding , Eric R. Hugo , Gila Idelman , Anuradha De Silva , Nathan W. Richtand , Jean 2 1 Loftus , Nira Ben-Jonathan * 1 Department of Cancer and Cell Biology, University of Cincinnati, Cincinnati, Ohio, United States of America, 2 The Christ Hospital, Cincinnati, Ohio, United States of America Abstract Introduction: Dopamine (DA) binds to five receptors (DAR), classified by their ability to increase (D1R-like) or decrease (D2R- like) cAMP. In humans, most DA circulates as dopamine sulfate (DA-S), which can be de-conjugated to bioactive DA by arylsulfatase A (ARSA). The objective was to examine expression of DAR and ARSA in human adipose tissue and determine whether DA regulates prolactin (PRL) and adipokine expression and release. Methods: DAR were analyzed by RT-PCR and Western blotting in explants, primary adipocytes and two human adipocyte cell lines, LS14 and SW872. ARSA expression and activity were determined by qPCR and enzymatic assay. PRL expression and release were determined by luciferase reporter and Nb2 bioassay. Analysis of cAMP, cGMP, leptin, adiponectin and interleukin 6 (IL-6) was done by ELISA. Activation of MAPK and PI3 kinase/Akt was determined by Western blotting. Results: DAR are variably expressed at the mRNA and protein levels in adipose tissue and adipocytes during adipogenesis. ARSA activity in adipocyte increases after differentiation. DA at nM concentrations suppresses cAMP, stimulates cGMP, and activates MAPK in adipocytes. Acting via D2R-like receptors, DA and DA-S inhibit PRL gene expression and release. Acting via D1R/D5R receptors, D