ERK1/2 pathway is involved in the inhibitory effect of crocetin on angiotensin II-induced vascular smooth muscle cell proliferation.
Angiotensin II (Ang II) induces vascular smooth muscle cells (VSMCs) proliferation, which plays an important role in the development and progression of atherosclerosis. Ang II-induced cellular events have been implicated, in part, in the activation of extracellular signal-regulated kinases 1/2 (ERK1/2). Crocetin is a natural carotenoid compound isolated from Gardenia jasminoids Ellis. In the present study, we investigated the effect of crocetin on the Ang II-induced VSMCs proliferation and ERK1/2 activation. 3-[4,5-dimethylthiazol-2-yl]-2,5-dephenyl tetrazolium bromide (MTT) and [3H]thymidine incorporation assay showed that the Ang II-induced VSMCs proliferation was inhibited significantly by crocetin. In-gel kinase assay indicated that Ang II elicited rapid and significant increase of ERK1/2 activity in VSMCs, which was suppressed by crocetin markedly. Western blotting analysis and cell-based enzyme-linked immunosorbent assay (ELISA) demonstrated that crocetin significantly inhibited the phosphorylation and activation of ERK1/2 induced by Ang II. Using the indirect immunofluorescent technique, we also found that crocetin inhibited nuclear translocation of activated ERK1/2 induced by Ang II. These findings suggest that the suppression by crocetin of the Ang II-induced VSMCs proliferation can be attributed, at least in part, to its inhibitory effect on ERK1/2 pathway.
Zhou
Cheng-Hua
CH
Department of Pharmacology, China Pharmaceutical University, 24 Tongjia Xiang, Nanjing 210009, PR China.
Qian
Zhi-Yu
ZY
Zheng
Shu-Guo
SG
Xiang
Min
M
D021581:Active Transport, Cell Nucleus / Q000187:drug effects; D000804:Angiotensin II / Q000494:pharmacology*; D000818:Animals; D000975:Antioxidants / Q000494:pharmacology; D015153:Blotting, Western; D002338:Carotenoids / Q000494:pharmacology*; D002417:Cattle; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D004305:Dose-Response Relationship, Drug; D004789:Enzyme Activation / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D019084:Fluorescent Antibody Technique, Indirect; D020935:MAP Kinase Signaling System / Q000187:drug effects; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism*; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010766:Phosphorylation / Q000187:drug effects; D014801:Vitamin A / Q000031:analogs & derivatives
D000975:Antioxidants; C487773:trans-sodium crocetinate; D014801:Vitamin A; D000804:Angiotensin II; D002338:Carotenoids; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3