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

作　　者：Ting-ting LIU Ti-long DING Yong MA Wei WEI 

机构地区：[1]Institute of Clinical Pharmacology, Anhui Medical University, Key Laboratory of Anti-inflammatory and Immune Medicine (Anhui Medical University), Ministry of Education, Hefei 230032, China [2]Department of Liver Diseases, 123th Hospital of PLA of China, Bengbu 233015, China

出　　处：《Acta Pharmacologica Sinica》2014年第11期1385-1392,共8页中国药理学报（英文版）

基　　金：Acknowledgements This study was supported by the National Natural Science Foundation of China （No 81173075） and the Nanjing Military Medical Science and Technology Innovation Foundation of China （No 10MA037）.

摘　　要：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 α1A-, α1B-, and （α1D-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-β1 （TGF-β1） and a-smooth muscle actin （α-SMA）], extracellular matrix （ECM） secretion factors [tissue inhibitor of metalloproteinase-1 （TIMP-1） and collagen-I （Coil）] and PKC-PβK-AKT signaling components （PKC, PI3K, and AKT） in the cells were detected by Western blotting and RT-PCR. Results： Both （α8- and α1B-AR were expressed in the membrane of HSC-T6 cells, whereas （α1B-AR was not detected. Treatment of the cells with NA concentration-dependently increased cell proliferation （EC50=277 nmol/L）, which was suppressed by the α1B-AR antagonist CEC or by the α1B-AR antagonist BMY7378. Furthermore, NA （0.001, 0.1, and 10 pmol/L） concentration-dependently increased the expression of TGF-β1, α-SMA, TIMP-1 and Coil, PKC and PI3K, and phosphorylation of AKT in HSC-T6 cells, which were suppressed by CEC or BMY7378, or by pertussis toxin （PT）, R0-32-0432 （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 Gα-coupled （Gα-AR and α1B-AR and the PKC-PI3K-AKT signaling pathway.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 α1A-, α1B-, and （α1D-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-β1 （TGF-β1） and a-smooth muscle actin （α-SMA）], extracellular matrix （ECM） secretion factors [tissue inhibitor of metalloproteinase-1 （TIMP-1） and collagen-I （Coil）] and PKC-PβK-AKT signaling components （PKC, PI3K, and AKT） in the cells were detected by Western blotting and RT-PCR. Results： Both （α8- and α1B-AR were expressed in the membrane of HSC-T6 cells, whereas （α1B-AR was not detected. Treatment of the cells with NA concentration-dependently increased cell proliferation （EC50=277 nmol/L）, which was suppressed by the α1B-AR antagonist CEC or by the α1B-AR antagonist BMY7378. Furthermore, NA （0.001, 0.1, and 10 pmol/L） concentration-dependently increased the expression of TGF-β1, α-SMA, TIMP-1 and Coil, PKC and PI3K, and phosphorylation of AKT in HSC-T6 cells, which were suppressed by CEC or BMY7378, or by pertussis toxin （PT）, R0-32-0432 （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 Gα-coupled （Gα-AR and α1B-AR and the PKC-PI3K-AKT signaling pathway.

关 键 词：NORADRENALINE hepatic fibrosis hepatic stellate cell Α1-ADRENOCEPTOR G. PKC PI3K AKT 

分 类 号：Q577[生物学—生物化学] Q55