title: Effect of crocetin on vascular smooth muscle cells migration induced by advanced glycosylation end products.
abstract: Crocetin is a major active constituent of Gardenia jasminoides J. Ellis, and can aid in the prevention of cardiovascular disease. The effect and possible mechanism of crocetin on the migration of vascular smooth muscle cells (VSMCs) induced by advanced glycosylation end products (AGEs) were investigated. VSMCs were pre-incubated with or without crocetin and exposed to AGEs subsequently. The invasion of the cells was investigated using a 24-well Cell Invasion Chamber. The anti-proliferative activity of crocetin was evaluated by MTT assay and VSMCs cell-cycle distribution was examined by flow cytometry. Cytokine TNF-α and IL-6 secreted by VSMCs and the amount of matrix metalloproteinase MMP-2 and MMP-9 in the culture supernatant were detected by ELISA. The expression level of RAGE (AGEs receptor), in cells was analyzed by western blot. The results demonstrated that AGEs increased about two-fold migration of VSMCs compared with control (OD=0.778±0.191 vs OD=0.413±0.214, P<0.01), and the proliferation increased by about 20% (OD=0.335±0.043 vs OD=0.281±0.037, P<0.01). Pre-treatment with crocetin (1.0μM) or RAGE antibody (10μg/ml) could inhibit the AGEs triggered migration of VSMCs obviously. Furthermore, both crocetin and RAGE antibody inhibited the increase of RAGE protein in VSMCs stimulated by AGEs. The levels of TNF-α and IL-6 decreased in the crocetin (1.0μM) pre-treated group compared to the AGEs (without pre-treated) group (37.60±3.08pg/ml vs 46.59±1.92pg/ml, 32.11±4.69pg/ml vs 49.99±8.84pg/ml, respectively). Crocetin (1.0μM) also reduced the value of MMP-2 and MMP-9 compared with the AGEs group (2.81±0.35ng/ml vs 6.40±0.85ng/ml, 2.69±0.25ng/ml vs 4.32±0.57ng/ml, respectively). In summary, crocetin inhibits the migration of VSMCs induced by AGEs through RAGE-dependent signaling pathway. And it is meaningful to diabetic vascular complications.
authors.affiliation: Department of Pharmacy, Suzhou Health College, Suzhou, Jiangsu 215009, China. Electronic address: xiangmin99@126.com.
authors.forename: Min
authors.initials: M
authors.lastname: Xiang
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Runlin
authors.initials: R
authors.lastname: Yang
authors.affiliation: Key Laboratory of Biotechnology for Analytical Medicine, Suzhou, Jiangsu 215009, China.
authors.forename: Yaqin
authors.initials: Y
authors.lastname: Zhang
authors.affiliation: Key Laboratory of Biotechnology for Analytical Medicine, Suzhou, Jiangsu 215009, China.
authors.forename: Pingping
authors.initials: P
authors.lastname: Wu
authors.affiliation: Key Laboratory of Nuclear Medicine, Ministry of Health, Jiangsu Key Laboratory of Molecular Nuclear Medicine, Jiangsu Institute of Nuclear Medicine, Wuxi, Jiangsu 214063, China.
authors.forename: Lizhen
authors.initials: L
authors.lastname: Wang
authors.affiliation: Department of Pharmacy, Suzhou Health College, Suzhou, Jiangsu 215009, China.
authors.forename: Zhenyu
authors.initials: Z
authors.lastname: Gao
authors.affiliation: Department of Pharmacy, Suzhou Health College, Suzhou, Jiangsu 215009, China.
authors.forename: Jianmei
authors.initials: J
authors.lastname: Wang
mesh_terms: D000818:Animals; D002338:Carotenoids / Q000494:pharmacology*; D002465:Cell Movement / Q000187:drug effects*; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D003925:Diabetic Angiopathies / Q000378:metabolism / Q000473:pathology / Q000517:prevention & control*; D004305:Dose-Response Relationship, Drug; D017127:Glycation End Products, Advanced / Q000633:toxicity*; D015850:Interleukin-6 / Q000378:metabolism; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D032389:Myocytes, Smooth Muscle / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D051381:Rats; D000067759:Receptor for Advanced Glycation End Products / Q000187:drug effects / Q000378:metabolism; D015398:Signal Transduction / Q000187:drug effects; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism; D014801:Vitamin A / Q000031:analogs & derivatives
chemical_list: C000593874:Ager protein, rat; D017127:Glycation End Products, Advanced; D015850:Interleukin-6; D000067759:Receptor for Advanced Glycation End Products; D014409:Tumor Necrosis Factor-alpha; C487773:trans-sodium crocetinate; D014801:Vitamin A; D002338:Carotenoids; D020778:Matrix Metalloproteinase 2; C522363:Mmp2 protein, rat; D020780:Matrix Metalloproteinase 9; C572337:Mmp9 protein, rat
keywords: AGEs; Crocetin; Diabetic vascular complications; Migration; Proliferation; VSMCs