Neferine inhibits angiotensin Ⅱ-stimulated proliferation in vascular smooth muscle cells through heme oxygenase-1  

作　　者：Xiao-chun LI Guo-xin TONG Yu ZHANG Shan-xin LIU Qi-hui JIN Huai-hong CHEN Peng CHEN 

机构地区：[1]Department of Geratology, the Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China [2]Department of Cardiology, the First People's Hospital of Hangzhou, Hangzhou 310006, China [3]Department of Cardiology, the First Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310003, China

出　　处：《Acta Pharmacologica Sinica》2010年第6期679-686,共8页中国药理学报（英文版）

摘　　要：Aim： To explore the effect of neferine on angiotensin Ⅱ （Ang Ⅱ）-induced vascular smooth muscle cell （VSMC） proliferation. Methods： Human umbilical vein smooth muscle cells （HUVSMCs） were used. Cell proliferation was determined by using the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay and flow cytometry analysis. Heme oxygenase （HO）-Ⅰ protein expression was tested by Western blot analysis. Extracellular signal-regulated protein kinase 1/2 （ERK1/2） activation was determined by using immunoblotting. Results： Pre-incubation of HUVSMCs with neferine （0.1, 0.5, 1.0, and 5.0 μmol/L） significantly inhibited Ang II-induced cell proliferation in a concentration-dependent manner and neferine 5.0 μmol/L increased HO-1 expression by 259% compared with control. The antiproliferative effect of neferine was significantly attenuated by coapplication of zinc protoporphyrin IX （ZnPP IX, an HO-1 inhibitor） with neferine. Ang Ⅱ-enhanced ERK1/2 phosphorylation was markedly reversed by neferine. By inhibiting HO-1 activity with ZnPP IX, the inhibitive effect of neferine on ERK1/2 phosphorylation was significantly attenuated. Cobalt-protoporphyrin （CoPP）, an HO-1 inducer, significantly decreased Ang II-induced ERK1/2 phosphorylation and inhibited Ang Ⅱ-induced cell proliferation. The ERK1/2 pathway inhibitor PD98059 significantly blocked Ang Ⅱ-enhanced ERK1/2 phosphorylation and inhibited cell proliferation. Conclusion： These findings suggest that neferine can inhibit Ang Ⅱ-induced HUVSMC proliferation by upregulating HO-1, leading to the at least partial downregulation of ERK1/2 phosphorylation.Aim： To explore the effect of neferine on angiotensin Ⅱ （Ang Ⅱ）-induced vascular smooth muscle cell （VSMC） proliferation. Methods： Human umbilical vein smooth muscle cells （HUVSMCs） were used. Cell proliferation was determined by using the 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyltetrazolium bromide （MTT） assay and flow cytometry analysis. Heme oxygenase （HO）-Ⅰ protein expression was tested by Western blot analysis. Extracellular signal-regulated protein kinase 1/2 （ERK1/2） activation was determined by using immunoblotting. Results： Pre-incubation of HUVSMCs with neferine （0.1, 0.5, 1.0, and 5.0 μmol/L） significantly inhibited Ang II-induced cell proliferation in a concentration-dependent manner and neferine 5.0 μmol/L increased HO-1 expression by 259% compared with control. The antiproliferative effect of neferine was significantly attenuated by coapplication of zinc protoporphyrin IX （ZnPP IX, an HO-1 inhibitor） with neferine. Ang Ⅱ-enhanced ERK1/2 phosphorylation was markedly reversed by neferine. By inhibiting HO-1 activity with ZnPP IX, the inhibitive effect of neferine on ERK1/2 phosphorylation was significantly attenuated. Cobalt-protoporphyrin （CoPP）, an HO-1 inducer, significantly decreased Ang II-induced ERK1/2 phosphorylation and inhibited Ang Ⅱ-induced cell proliferation. The ERK1/2 pathway inhibitor PD98059 significantly blocked Ang Ⅱ-enhanced ERK1/2 phosphorylation and inhibited cell proliferation. Conclusion： These findings suggest that neferine can inhibit Ang Ⅱ-induced HUVSMC proliferation by upregulating HO-1, leading to the at least partial downregulation of ERK1/2 phosphorylation.

关 键 词：NEFERINE heme oxygenase-1 angiotensin Ⅱ zinc protoporphyrin vascular smooth muscle cells cell proliferation 

分 类 号：Q55[生物学—生物化学] Q463