title: Bisoprolol reverses epinephrine-mediated inhibition of cell emigration through increases in the expression of β-arrestin 2 and CCR7 and PI3K phosphorylation, in dendritic cells loaded with cholesterol.
abstract: The effect of bisoprolol on dendritic cell (DC) migration was investigated, including the analysis of protein expression, cytokine secretion and activation of the PI3K-pathway. The chemotactic cell numbers in cholesterol-loaded DCs treated with epinephrine were significantly decreased by 26.66±6.29% (6h), 35.67±2.91% (12h) and 29.33±1.09% (24h). This effect was significantly reversed by 46.00±10.65% (6h), 64.25±6.77% (12h) and 55.74±5.51% (24h) when bisoprolol and epinephrine were both present. In cholesterol-loaded DCs, treatment with epinephrine significantly increased AR-β1 protein expression by 56.99±4.87%, but inhibited β-arrestin 2 and CCR7 protein expression by 30.51±4.22% and 25.31±0.04%, respectively. These effects were reversed by bisoprolol by 36.87±4.40%, 41.47±3.95% and 30.14±0.54%, respectively. TNF-α and MMP9 levels were decreased by 68.33±4.00% and 39.57±9.21% in cholesterol-loaded DCs treated with epinephrine. In contrast, when bisoprolol and epinephrine were administered together, the secretion of these proteins was significantly increased by 233.81±37.06 % and 76.66±14.21%, respectively. Treatment with epinephrine decreased PI3K-phosphorylation by 31.88±2.79%, 40.24±5.69% and 30.93±4.66% at 15, 30 and 60min, respectively, whereas the effect of epinephrine on the expression of phosphorylated PI3K was reversed by 49.49±12.12%, 70.93±16.14% and 47.62±6.00%, respectively, when cells were treated with both bisoprolol and epinephrine. Wortmannin inhibited the effects of bisoprolol on PI3K-phosphorylation (38.63±6.12%), the expression of CCR7 (23.4±2.72%), the secretion of TNF-α (69.46±4.48%) and MMP9 (43.15±4.63%), and the number of chemotactic cells (36.84±5.22%). This is the first study to establish a signaling pathway, epinephrine-AR-β1-β-arrestin2-PI3K-MMP9/CCR7, which plays a critical role in the migration of DCs.
authors.affiliation: Division of Biochemistry, Tongji University School of Medicine, Shanghai 200092, China. hyang@tongji.edu.cn
authors.forename: Hong
authors.initials: H
authors.lastname: Yang
authors.forename: Rong-Zeng
authors.initials: RZ
authors.lastname: Du
authors.forename: Jian-Ping
authors.initials: JP
authors.lastname: Qiu
authors.forename: Yong-Qing
authors.initials: YQ
authors.lastname: Tang
authors.forename: Shi-Chao
authors.initials: SC
authors.lastname: Chen
mesh_terms: D058671:Adrenergic beta-1 Receptor Antagonists / Q000494:pharmacology; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000730:Androstadienes / Q000494:pharmacology; D019390:Arrestins / Q000378:metabolism*; D050197:Atherosclerosis / Q000378:metabolism; D017298:Bisoprolol / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects; D002633:Chemotaxis / Q000187:drug effects*; D002784:Cholesterol / Q000378:metabolism; D003713:Dendritic Cells / Q000166:cytology / Q000187:drug effects*; D004837:Epinephrine / Q000494:pharmacology*; D005434:Flow Cytometry; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation; D054400:Receptors, CCR7 / Q000378:metabolism*; D013997:Time Factors; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D000077191:Wortmannin; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins
chemical_list: C000605254:ARRB2 protein, human; D058671:Adrenergic beta-1 Receptor Antagonists; D000318:Adrenergic beta-Agonists; D000730:Androstadienes; D019390:Arrestins; C515370:CCR7 protein, human; D054400:Receptors, CCR7; D014409:Tumor Necrosis Factor-alpha; D000071560:beta-Arrestin 2; D000071557:beta-Arrestins; D002784:Cholesterol; D019869:Phosphatidylinositol 3-Kinases; C579270:MMP9 protein, human; D020780:Matrix Metalloproteinase 9; D000077191:Wortmannin; D017298:Bisoprolol; D004837:Epinephrine