Effects of roscovitine on proliferation and cell cycle induced by TNF-α of vascular smooth muscle cells in rats / 中国药理学通报

ABSTRACT

Aim: To observe the effect of roscovitine on TNF-α-induced proliferation and cell cycle of rat vascular smooth muscle cells (VSMCs). Methods: VSMCs were isolated and cultured using tissue explant method. Tumor necrosis factor-alpha (TNF-α) was used to induce the proliferation of VSMCs. VSMCs were divided into control group, TNF-α group and roscovitine 5, 10, 15, 30 μmol · L-1 group. Cell proliferation was detected with MTT, and PCNA expression was tested. Flow cytometry was used to detect the cell cycle. The expressions of cyclin proteins (Cyclin A, Cyclin B, Cyclin D, Cyclin E), cyclin-dependent kinase proteins(CDK4, CDK5), and cell cycle inhibitory proteins(p53, p21, p27) were examined by realtime reverse transcription PCR(qRT-PCR) and Western blot. Results: Roscovitine could inhibit the proliferation of VSMCs. Roscovitine inhibited transformation of cell cycle from G0/G1 phase to S phase. Compared with TNF-a group, the expressions of Cyclin A, Cyclin B, Cyclin D, Cyclin E protein decreased, the expressions of CDK4 and CDK5 protein decreased, while the expression of p53, p21 and p27 protein in cell cycle increased of roscovitine 5, 10, 15, 30 μmol · L-1 group (P < 0. 05). Conclusion: Roscovitine inhibits the cell cycle progression and proliferative activity of rat VSMCs.