The Expression Change Of Slit/Robo In ECFC From Diabetic And In HUVEC Induced By Hyperglycemia And Hyperlipemia

Objective:To observe the expression change of Slit/Robo in ECFC from diabetic.Methods:Mononuclear cells were isolated from fasting venous peripheral blood of diabetic and healthy adult by density gradient centrifugation and cultured with EGM-2medium until the colony cells were formed. Cell phenotype were analyzed by flow cytometry analysis and the high,medium and low colony cells were isolated,identified and counted. The mRNA expression of Slit/Robo were detected by real-time PCR.Results:(1)The99.24%of ECFC expressed CD31and the expression of CD45and CD14were negative,indicating that the ECFC were successfully isolated and cultured.(2)The percentage of high colony cells in ECFC from diabetic was lower than that of healthy adult(p<0.05).(3)The mRNA expression of Slit2,Slit3,Robol and Robo4in ECFC from diabetic were0.12、0.49、0.19、0.11times of healthy adult respectively and the mRNA expression of Slit2and Robo4in ECFC from diabetic were significantly decreased(p<0.05).Conclusions:The proliferation of colony cells and the expression of Slit2and Robo4were decreased in ECFC from diabetic compared with healthy adult.Figures7,References32. AbstractObjective:To observe the expression change of Slit/Robo in HUVEC induced by hyperglycemia and hyperlipemia.Methods:HUVEC were cultured in RPMI1640medium and treated with standard or high glucose for ld,2d,3d,5d and7d, the mRNA and protein expression of Slit/Robo were detected by real-time PCR and Western blot analysis respectively. HUVEC were devided into control and hyperlipemia and treated with0,50,125,250μmol/L hexadecylic acid in RPMI1640medium for48h, the mRNA expression of Slit/Robo were detected by real-time PCR.Results:(1)The mRNA expression of Slit2,Slit3and Robo4were increased in HUVEC in the second,third and fifth day treated with high glucose.(2)The protein expression of Slit2,Slit3and Robo4were increased in HUVEC treated with high glucose especially Slit3and Robo4increased in the second,third and fifth day and Slit2increased in the second,third,fifth and seventh day.(3)The mRNA expression of Slit2,Slit3and Robol were increased in HUVEC treated with125μmol/L hexadecylic acid and the mRNA expression of Robo4were increased in HUVEC treated with250μmol/L hexadecylic acid.Conclusions:High glucose could promote the expression of Slit2,Slit3and Robo4in HUVEC;hyperlipemia could promote the expression of Slit2,Slit3, Robo1and Robo4in HUVEC. Figures14peferen(?)32...