Effects Of Tanshinone ⅡA On Proliferation In Rat Vascular Smooth Muscle Cells And The Expressions Of P21 Protein In Angiotensin Ⅱ-induced

Objective: For FBS is basic alimentation ,to observe the effects of tanshinoneâ…¡A (TSN)on proliferation and apoptosis of vascular smooth muscle cells(VSMC) induced by angiotensinâ…¡(AngII), and explore its relationship with cell cycle protein p21.Methods:(1)Sprague-Dawley(SD) rats, aorta VSMC with the tissue method, using passage 4-6 cells . Before the beginning of the experiment, in the follow-up immunohistochemical method using cultured cells is the identification of VSMCs. Then let the cells without of FBS in the DMEM culture medium for 48 hours, in order to enable synchronization between cells in G0 phase;(2)Interfere with cell division by the experimental:control group (DMEM medium containing 5% FBS); normal control group(DMEM medium containing 5% FBS +1μmol / LAngII); experimental group (DMEM medium containing 5% FBS +1μmol / LAngII + different concentrations of TSN);(3)The use of MTT method assay TSN cytotoxicity to VSMCs, and find out inhibition rate of half (IC₅₀), then using the same method-MTT assay to acquire of the TSN on the AngII-induced inhibition of VSMC proliferation;(4)Choice of TSN in the vicinity of the role of IC₅₀ concentration, flow cytometry for apoptosis rate of VSMCs, scanning electron microscopy morphology of VSMCs apoptosis;(5)Using RT-PCR to detecting the RNA expression of p21, then using cell immunohistochemistry method to detecting the protein expression of p21. Results:(1)Tanshinone II-A (TSN) in the IC₅₀ range can inhibit the inducing by angiotensinâ…¡(AngII),the in vitro cultured rat vascular smooth muscle cell proliferation, the concentration of intergroup and control group statistically significant differences (p<0.01) and with the concentration increase, a marked increase in inhibition. This inhibitory effect of p21 have cell cycle protein expression, and showed a dose-dependent manner (p< 0.05).(2)Tanshinone II-A (TSN) in the range of IC₅₀, with the increase in concentration can occur with the control group, there are significant differences in the role of cell apoptosis (p<0.05), and the emergence of the cell cycle protein expression of p21.Conclusion :(1)TSN range in IC₅₀ can cause significant inhibition of proliferation of VSMCs, and even apoptosis.(2)TSN in the IC₅₀ range of VSMCs caused by inhibition of pro- liferation and apoptosis may be related to the expression of cell cycle protein p21-related enhancements.