Strengthen Our Foundation The Phlegm Tongmai Side Containing Serum On Rat Vsmc Proliferation And The Of Cyclind <sub> 1 </ Sub> Affect Experimental Research

ObjectiveTo study the proliferation law of platelet-derived growth factor BB (PDGF-BB)stimulation on the vascular smooth muscle cells (VSMC) and establish the cellproliferation modle. Investigate the influence and mechanism of thecontained-GHTR(Guben Huatan Tongmai Recipe) serum on the VSMC of rat in thecell proliferation, cell cycle and the expression of cyclin D₁ with the method of serumphamcology.MethodsBoiled and filtered the GHTR decoction, garaged wistar rat with certain dose.Accoding to the method of serum pharmacology, extracted the contained-GBTRserum. Selected several male wistar rats, exposed their main arteries of thorax. Chosethe middle membrane for primary cell culture. The proliferation models of vascularsmooth muscle cells were estabilished and used for the experiment when it grew atfour generations. Divided cells into 4 groups at random: Normal group, PDGF-BBstimulation group(Model group), high dose of contained-GHTR serum and low doseof contained-GHTR serum. All the group stimulated with PDGF-BB except thenormal group for 24h to improve the cell proliferation, then added different dose ofcontained-GHTR serum for another 24h. The normal group and the model group weretreated with blank serum from rat. Test the suppress ratio of the contained-GHTRserum on the VSMC with MTT. Observed cell cycle and the express of cyclinD₁ withFCM.Results1.The primary culture of the VSMC was successful. The VSMC grew well. Itspurity and amount fit for the demand of experiment. Cells were spindle or longspindle, radioactive growth, cell arrange parallel beam, part of the regional multi-celloverlapping part of the district was single, undulating, VSMC showed the typical "peak" "Ceres" shape growth. When cell reproduced at 3 generation, cells wereidentificated with the anti—actin immunohistochemistry, vascular smooth muscle cellspositive rate＞95%. 2. Stimulated with PDGF-BB and intervened withcontained-GHTR serum, both the high dose of contained-GHTR and the low dose ofcontained-GHTR could suppress the impoving proliferation effect of PDGF-BB onthe VSMC. They had significant statistical difference with the modle group in theoptical density. 3. Flow cytometry showed that PDGF-BB can improve the cell cycleand increase the expression of cyclin D₁, it had significant difference with the normalgroup; GHTR inhibited VSMC from inactive to the synthesis phase, so that the cellcycle of VSMC arrested in the G₀/G₁ phase, and downregulated the expression ofcyclin D₁.Conclusion1. PDGF-BB can promote the proliferation of VSMC. GHTR significantlyinhibited the stimulation of PDGF-BB on VSMC proliferation, it has the function ofpreventing and curing atherosclerosis (AS). 2. GHTR inhibit cell proliferation cycle, itcan significantly inhibit VSMC from inactive to the synthesis phase and make the cellcycle arrest in the G₀/G₁ phase. 3. PDGF-BB can increase the expression of cyclin D₁, but GHTR can obviously reduce the expression of cyclinD₁, it suggested that GHTRcontrol mechanism may be related to its regulation of vascular smooth muscle cellscyclin D₁ activity.