Effects Of Hydroxylcamptothecinum. Aclarubicin. Triptolide On The Cell Cycle Of Cultured Vascular Smooth Muscle Cells

OBJECTIVETo assess the effects of Hydroxylcamptothecinum(HCPT), Triptolide(TP) and Aclarubicin(ACR) involved in proliferation and apoptosis of vascular smooth muscle cells, and explore their relationship of concentration and time-dependence, and to screen a new drug for the drug-eluting stents.METHODSThe New Zealand rabbit aortic VSMCs were cultured and identified with optical microscopy (OM) and electric microscopy (EM).First, cultured cells were incubated with HCPT, TP and ACR at different concentrations for 72 hours to evaluate the growth effects of HCPT, TP and ACR on VSMCs and to determine the optimal experimental conditions. Second, cells were divided into the following five groups: 1×10-4M HCPT,1×10-5M TP, 5×10-6M ACR,1×10-4M DXM (the positive control) and IMDM group(the negative control) to characterize these three drugs involved in the above effect, in which cultured cells were incubated for 1.3 and 7 days.3. Cell proliferation was measured with cell counting techniques, WST-1 enzyme-linked immunosorbent assay (ELISA), cell apoptosis was evaluated by flow cytometry(FCM), agarose gel electrophoresis assay and electronic microscopy.RESULTS1. VSMCs cultured from the rabbit thoracic aorta showed the characterizatic of VSMCs including spindle shaped cells arranged in an interwoven and a peak and valley pattern.After treated with different concentration of HCPT, TP and ACR for 1, 3 and 7 days, VSMCs number, metabolic activity (WST-1 cleavage)decreased significantly in a manner of time-dependence(P<0.01),and TP dramatically inhibited the proliferation of VSMCs.After treated with 1×10-4M HCPT, 1×10-5M TP, 5×10-6M ACR, 1×10-4M DXM for 1, 3 and 7 days, the apoptotic peak in those groups elevated. The apoptotic rate of 1×10-5M TP group was significantly higher than that in DXM group (P<0.01).4. DNA ladders were also found in the treated groups, VSMCs apoptotic ultrastructure changes confirmed by electric microscopy.CONCLUSIONSThe study demonstrates that HCPT, TP and ACR inhibit proliferation and induce apoptosis in VSMCs, and TP shows a more promising role.