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To investigate whether rosuvastatin improve reendothelialization of late endothelial progenitor cells (EPCs) induced by C reactive protein (CRP) through the advanced glycation end products (RAGE)/eNOS signaling pathway.
1.Human late EPCs were treated with CRP of different concentrations (0, 10, 25, 50ug/mL) to explore the CRP-induced RAGE/ eNOS signaling pathway. 2.EPCs were pre-incubated with rosuvastatin of different concentrations (0, 10-8M, 10-7M, 10-6M) and then stimulated with 50ug/mL CRP, with EPCs incubated with 50ug/mL CRP as positive control. The mRNA levels of eNOS for EPCs were measured by quantitative PCR. NO secretions were detected by NO nitrate reductive enzymatic. RAGE protein expressions were checked by western blotting. MTT assay was used to test Proliferation of EPCs, transwell assay for migration, and adhesiveness assay for adhesion.
1.When EPCs were incubated with CRP at different dosages, the expression of eNOS mRNA and NO secretions dropped gradually compared with normal control (P <0.05). In contrast, RAGE protein expressions increased in a dose-dependent manner (P <0.05). 2. The RAGE proteins expressions in positive control raised to 121%±4% (P <0.01) when compared with normal control. After pre-incubated with increasing concentrations of rosuvastatin, RAGE proteins expressions declined gradually to 115%±10%, 110%±10% and 87%±4% (P <0.01). The eNOS mRNA level in positive control declined to 44%±19% when compared with normal control, and raised to 58%±13%, 82%±32% and 203%±52% (P <0.01) with increasing concentration of rosuvastatin. Similarly, NO secretions in positive control significantly declined compared with normal control, and up-regulated gradually in a dose-dependent manner when treated with rosuvastatin. Simultaneously, the migration and adhesion of CRP-induced EPCs were up-regulated along with increasing concentration of rosuvastatin. However, the proliferation of EPCs had no changes.
CRP could influence expressions of eNOS/NO though signaling pathway mediated by RAGE. Rosuvastatin improves EPCs’ reendothelialization induced by CRP though restraining RAGE.