Effects Of Endogenous Carbon Monoxide On Gene Expression Pattern Associated With Proliferation And Apoptosis Of Pulmonary Arterial Smooth Muscle Cell

Objective To study the gene expression associated with proliferation and apoptosis of pulmonary arterial smooth muscle cells stimulated shortly by endogenous carbon monoxide (CO) and PDGF and speculate on the possible underlying mechanisms with microarray approaches.Methods After male SD rats were anesthetized, pulmonary arterial smooth muscle cells were cultured in vitro. They were confirmed by morphology and immunohistochemical stains ofα-actin. They were divided randomly into 3 groups: GroupⅠor normal control group; GroupⅡor group stimulused by PDGF (concentration: 20ng/ml); GroupⅢor group stimulused by PDGF(concentration: 20ng/ml) and Hemin(concentration: 20μmol/L) (respectively 5 culture plates), cells treated with PDGF and Hemin were incubated for 2 hours, Total RNA was isolated, quantitated and purified. CDNA was amplified by RT-PCR from total RNA. Next,biotin labeled CRNA obtained by in vitro transcription was quantitated and purified. After hybridisation, microarray was washed, stained, scanned and analyzed by computer system. Result Some of genes were upregulated such as CyclinD,CyelinH1,CyclinL1,MAP2K3 (P38),Mybll,Junb,Jun,Kras,Nras,VEGF-α,FGF-α,PDGF-α,Rags,Rbm7,Socs,Bzwl,Eif4e and etc in groupⅡcompared to control,, but P27,Nek2,Gadd45α,Macfl,Trp53inp1,Parc ete were significantly downregulated. GroupⅢCompared with groupⅡ, genes such as MAP2K3 (P38),CyclinD1,CyclinG1,GOS2,TGFb3,PDGF-α,IGF1,MybL1,anape10,Faim,Pik3r1 were significantly downregulated,while Gadd45α,P21,Sens2,Trp53inpl,sqstml,Ernl etc genes were markedly upregulated. Genes such as P21,Gadd45α,Trp53inp1,sqstm1,Ccar1 erc were significantly upregulated in groupⅢcompared to control.Conclusions We speculated that proliferation were stimulated via Map2k3 signal transduction and a series of genes expression such as growth factors,oncogenes and cyclins, when pulmonary arterial smooth muscle cells treated shortly with PDGF. Map2k3 signal pathway may be also involved in growth arrest led by endogenous carbon monoxide. Moreover, the results suggested that P53 pathway probably play an important role in apoptosis of pulmonary arterial smooth muscle cells treated with endogenous carbon monoxide Meanwhile, Caspases family and Gadd45αmay participate in it. We have seen that the proliferation and apoptosis of pulmonary arterial smooth muscle cells were complicated process with many signal transductions and many genes. The results disclosed in microarray would guide our further investigation.