Study On The Effect Of Angiotensin Converting Enzyme Antisense CDNA On Vascular Smooth Muscle Cells And Cardiac Fibroblasts

Object: A vector containing angiotensin converting enzyme antisense sequence (ACE-AS) was used to determine the feasibility of gene therapy mediated by liposome to attenuate ACE activity in vascular smooth muscle cells (VSMCs) and cardiac fibroblasts(Fbs) from SHR and WKY rats.To investigate the effect of angiotensin converting enzyme antisense cDNA on the proliferation of vascular smooth muscle cells from SHR and WKY rats, and the machanism of antisense cDNA.To investigate the effect of ACE antisense cDNA on the collagen synthesis of Fbs.Methods:In Vitro transfer VSMCs and Fbs from SHR and WKY rats with target ACE antisense cDNA(AS),ACE tuncated cDNA(TS) and empty vector(control) mediated by liposome. Two days aftertransferation,evaluates the target gene expression and the effect on ACE expression with RT-PCR, tests the target ACE activity of the cells with test reagent,and the Ang II level of the supernatant and cells with radioimmunoassay. Depicts the growth curve and calculates the double time of VSMCs. Test the DNA synthesis with 3H-Thymide incorporation rate in VSMCs from SHR and WKY.Test the collagen synthsis with 3H-Proline incorporation in Fbs.Results: ACE antisense cDNA can expressed in two kinds of cells,the expression peak was in the fourth days after transferation,and it wasconcomitant with the decrease of ACE expression. AS group has significantly lower 3H-Thymide incorporation rate in VSMCs from SHR than TS and control group(5993. 3 ?78. 3 cpm/105 cells vs!1449. 3?99. 5 , 11436. 3+2243. 4 cpm/105 cells P<0. 001) . This was associated with a significant decrease in ACE activity and Ang II level in target cells and culture medium.However, there is not same effect on VSMCs from WKY. AS group has significantly lower 3H-Proline incorporation rate in Fbs from SHR than TS and control group(4355. 7 + 75. 9 vs 6310. 3+98. 9 , 6529. 5 + 192. 5cpm/105 cells, PConclusion: ACE antisense cDNA can inhibite the abnormally higher basal DNA synthesis in SHR VSMCs and collagen synthesis in SHR Fbs. Present founding suggests that ACE antisense cDNA will be useful and possible in the gene therapy of arterial proliferative disease and cardiovascular remodeling.