Selective α1B- and α1D-adrenoceptor antagonists suppress noradrenaline-induced activation, proliferation and ECM secretion of rat hepatic stellate cells in vitro

Abstract

Aim: To explore the effects of noradrenaline (NA) on hepatic stellate cells (HSCs) in vitro and to determine the adrenoceptor (AR) subtypes and underlying mechanisms. Methods: The distribution and expressions of α 1A -, α 1B -, and α 1D -ARs in HSC-T6 cells were analyzed using immunocytochemistry and RT-PCR. Cell proliferation was evaluated with MTT assay. The expression of HSC activation factors [transforming factor-β 1 (TGF-β 1 ) and α-smooth muscle actin (α-SMA)], extracellular matrix (ECM) secretion factors [tissue inhibitor of metalloproteinase-1 (TIMP-1) and collagen-Ι (ColΙ)] and PKC-PI3K-AKT signaling components (PKC, PI3K, and AKT) in the cells were detected by Western blotting and RT-PCR. Results: Both α 1B -and α 1D -AR were expressed in the membrane of HSC-T6 cells, whereas α 1A -AR was not detected. Treatment of the cells with NA concentration-dependently increased cell proliferation (EC 50 =277 nmol/L), which was suppressed by the α 1B -AR antagonist CEC or by the α 1D -AR antagonist BMY7378. Furthermore, NA (0.001, 0.1, and 10 μmol/L) concentration-dependently increased the expression of TGF-β 1 , α-SMA, TIMP-1 and ColΙ, PKC and PI3K, and phosphorylation of AKT in HSC-T6 cells, which were suppressed by CEC or BMY7378, or by pertussis toxin (PT), RO-32-0432 (PKC antagonist), LY294002 (PI3K antagonist) or GSK690693 (AKT antagonist). Conclusion: NA promotes HSC-T6 cell activation, proliferation and secretion of ECM in vitro via activation of G α -coupled α 1B -AR and α 1D -AR and the PKC-PI3K-AKT signaling pathway.

Evidence for a direct effect of the autonomic nervous system on intestinal epithelial stem cell proliferation

“…NE and ACh released from these terminals may bind to receptors on the stem cells and/or the TA cells to affect proliferation. Both NE and ACh have been shown to directly alter proliferation in other cell types, mediated through a variety of adrenoreceptor and muscarinic receptor subtypes (Cruise et al 1985;Geloen et al 1988;Bronnikov et al 1992;Oben et al 2003a,b;Liu et al 2014Liu et al , 2017Sloniecka et al 2015;Morgan et al 2016). However, in contrast to other organ systems, the gastrointestinal (GI) tract is innervated by the enteric nervous system (ENS).…”

“…,b; Liu et al. , ). Twenty‐four hours after treatment administration, Matrigel was dissolved using 1 mL QIAzol (QIAGEN; Germantown, MD), the cells vortexed, and stored at −80°C to await processing (see Quantitative real time polymerase chain reaction).…”

“…The physiological neurotransmitters of the ANS were used instead of pharmacological agents targeting specific autonomic neurotransmitter receptors in order to most closely mimic the in vivo physiology of this system. Doses were chosen based on other studies demonstrating NE-or AChinduced proliferation in other cell types in vitro (Cruise et al 1985;Oben et al 2003a,b;Liu et al 2014Liu et al , 2017. Twenty-four hours after treatment administration, Matrigel was dissolved using 1 mL QIAzol (QIAGEN; Germantown, MD), the cells vortexed, and stored at À80°C to await processing (see Quantitative real time polymerase chain reaction).…”

Norepinephrine-stimulated HSCs secrete sFRP1 to promote HCC progression following chronic stress via augmentation of a Wnt16B/β-catenin positive feedback loop

“…Electron-microscopic studies have shown that HSCs, the major fibrogenic cells in the liver, are in contact with nerve fibers in the human liver [22]. In addition, several neuronal genes, including adrenergic receptors, are expressed in HSCs [48], and catecholamines are known to aggravate stress-induced liver diseases (e.g., cirrhosis) [49]. In this study, sFRP1 was chosen as the central molecule for the following reasons.…”

Indolylpiperidine derivatives as potent and selective α1B adrenoceptor antagonists

“…9,10 In addition, chloroethylclonidine as an irreversible a 1B adrenoceptor antagonist and pyrrolidin-2-one derivatives as selective a 1B adrenoceptor antagonists have been reported. 11,12 In the present study, however, we aimed to use other structural motifs for designing potent and selective a 1B adrenoceptor antagonists. In this Letter, we report the synthesis and structure-activity relationship of indole derivatives that show high affinity and selectivity for the a 1B adrenoceptor.…”