Establishment Of An Organic And Cellular Model Of VSMC Proliferation

Objective To explore an organic and cellular model of vascular smooth muscle cell (VSMC) proliferation and establish a favorable experimental platform for the study of proliferous vascular diseases, the aorta segments of rat were incubated in culture flask containing 20% bovine serum DMEM culture medium and VSMC from cultured aorta of rat was cultured with 20% bovine serum DMEM culture medium. The proliferous cells were labeled by Bromodeoxyuridine(BrdU) with immunocytochemical method, and the mRNA expressions of hypertension-related gene-1(HRG-1) and smooth muscle 22 alpha(SM22α) were detected by RT-PCR.Methods The organic model: 19 male rats between 150-200g were anaesthetized with chloral hydrate. The aortas were carefully dissected and removed the redundant adipose. Then the arteries were cut into segments of about 1.5cm. The endothelium of artery segments was injured with toothpick before being cultured. The artery segments were randomly divided into three experimental groups of cultured 5, 8 and 13 days respectively, and one control group in which the artery segments had not been cultured in vitro. The artery segments of the three experimental groups were incubated in 20% fetal bovine serum(FBS) DMEM at 37℃, 5% CO2 . BrdU was added into the culture medium before 24h of accomplishing samples. The cellular model: VSMCs from aorta cultured for 8 days in vitro and normal arteries were cultured respectively. Cells from passages 3-4 generations were used for the present experiment. BrdU was added into the medium before 24h of accomplishing cells. In the above experiments, The proliferous cells were observed with HE staining and were labeled by BrdU with immunocytochemical method, the microcosmic structure of VSMC was observed through transmission electron microscope(TEM), and the mRNA expressions of HRG-1 and SM22αwere detected by RT-PCR.