Identification of proteins involved in insulin stimulation in v ascular smooth muscle cells of SHR rats

AIM:To identify proteins involved in insul in stimulation and the molecular mechanism of proliferation and migration in vas cular smooth muscle cells (VSMCs).METHODS:A series of methods,including 2-D electrophoresis,PDQ uest software analysis of 2-DE gels,peptide mass fingerprinting based on matrix -assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TO F-MS) and SWISS-PROT database searching,were used to separate and identify the differentially expressed proteins.The difference of some proteins was proved by Western blotting.Proliferation and migration of VSMCs treated with insulin wer e also observed.RESULTS:DNA synthesis were increased in VSMCs.［3H］-thymidi ne incorporation in VSMCs from SHR (14.40±0.85) was higher than that in VSMCs from WKY (9.21±0.93,P<0.05).Migration rate of VSMCs from SHR was abou t 1.8 times higher than that in VSMCs from WKY (P<0.05).Image analysis re vealed that averages of protein spots detected were 502±32 and 612±39 in contr ol VSMCs and in cells treated with insulin,respectively.Result of western blo tting confirmed that α-SM protein was down-regulated and matrix Gla,OPN protei ns were up-regulated by insulin stimulation.CONCLUSION:The results suggest that the differential proteomic analysis may be useful to study related proteins involved in insulin-regulated p roliferation in VSMCs.