title: Morphine-mediated alteration of hypertension-related gene expression in human white blood cells and multilineage progenitor cells.
abstract: It has been shown that vascular endothelial cells functionally express a local circuit autocrine-paracrine regulatory pathway driven by endogenously expressed chemically authentic morphine, its cognate opiate alkaloid-selective mu3 and mu4 receptors, and constitutive nitric oxide (NO). Accordingly, the aim of the study was to examine morphine-mediated changes in hypertension-associated gene expression in two independent cell models: primary cultures of human white blood cells (WBCs) and human multilineage progenitor cells (MLPCs). In separate incubations, primary cultures of human WBC and MLPC were treated with morphine at a final concentration of 1  μM morphine for 2-4  h. After RNA extraction and reverse transcription, Human Genome Survey Arrays were used to construct and differentially analyze by strict statistical criteria transcriptional/gene expression profiles of WBC and undifferentiated human MLPC in three independent experiments. The Applied Biosystems Human Genome Survey Array contains 31,700 60-mer oligonucleotide probes representing a set of 27,868 individual human genes and >1000 control probes. After DNA microarray analyses, a variety of hypertension-associated genes from both cell types were observed to be significantly downregulated. The only genes expressed in both cell types were β-adrenergic receptor kinase 2 (ADRBK2) and coding protein kinase WNK1 (PRKWNK1); however, only PRKWNK1 showed downregulation of its expression after morphine exposure. Only two genes were observed to be significantly upregulated after morphine treatment: ADRBK2 in stem cells and β3-adrenergic receptor in WBC. Morphine administration to primary cultures of human WBC and MLPC altered the expression profile of 16 candidate hypertension-associated genes. The majority of relevant genes was observed to be downregulated, suggesting ongoing homeostatic regulation by endogenous morphine coupled to NO production and release. In sum, these data suggest a predominantly antihypertensive role for endogenous morphine/NO signaling events.
authors.affiliation: Department of Hypertension, Chair of Nephrology and Hypertension, Medical University of Lodz, Lodz, Poland.
authors.forename: M
authors.initials: M
authors.lastname: Banach
authors.forename: F M
authors.initials: FM
authors.lastname: Casares
authors.forename: R M
authors.initials: RM
authors.lastname: Kream
authors.forename: A
authors.initials: A
authors.lastname: Gluba
authors.forename: J
authors.initials: J
authors.lastname: Rysz
authors.forename: G B
authors.initials: GB
authors.lastname: Stefano
mesh_terms: D019070:Cell Lineage; D002478:Cells, Cultured; D054770:G-Protein-Coupled Receptor Kinase 3 / Q000235:genetics; D020869:Gene Expression Profiling / Q000379:methods; D005786:Gene Expression Regulation / Q000187:drug effects*; D006801:Humans; D006973:Hypertension / Q000097:blood / Q000235:genetics* / Q000503:physiopathology / Q000517:prevention & control; D047908:Intracellular Signaling Peptides and Proteins; D007962:Leukocytes / Q000187:drug effects* / Q000378:metabolism; D015778:Minor Histocompatibility Antigens; D009020:Morphine / Q000494:pharmacology*; D009569:Nitric Oxide / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D018847:Opioid Peptides / Q000378:metabolism; D017346:Protein Serine-Threonine Kinases / Q000235:genetics; D012333:RNA, Messenger / Q000097:blood; D022702:Receptors, Adrenergic, beta-3 / Q000235:genetics; D013234:Stem Cells / Q000187:drug effects* / Q000378:metabolism; D013997:Time Factors; D000076204:WNK Lysine-Deficient Protein Kinase 1
chemical_list: D047908:Intracellular Signaling Peptides and Proteins; D015778:Minor Histocompatibility Antigens; D018847:Opioid Peptides; D012333:RNA, Messenger; D022702:Receptors, Adrenergic, beta-3; D009569:Nitric Oxide; D009020:Morphine; D017346:Protein Serine-Threonine Kinases; D000076204:WNK Lysine-Deficient Protein Kinase 1; C498743:WNK1 protein, human; D054770:G-Protein-Coupled Receptor Kinase 3; C517848:GRK3 protein, human