Effects Of Pyridoxamine And Telmisartan On Proliferation And Migration Of Rat Vascular Smooth Muscle Cells Induced By Angâ…¡

ObjectiveThis study is designed to investigate the influence of pyridoxamine and telmisartan on the proliferation and migration of vascular smooth muscle cells (VSMCs) from Spontaneously Hypertensive Rat(SHR) induced by angiotensinⅡ(AngⅡ) and to elucidate the underlying mechanisms.MethodsThe culture of SHR thoracic aorta vascular smooth mucle cells was done by tissue-piece inoculation and the cells from the 3th to 5th passage were selected for the experiments.The cultured VSMCs were randomly divided into control group,AngⅡgroup(10~-7mol/L AngⅡ),P group(1mmol/L pyridoxamine+10~-7mol/L AngⅡ),T group(10~-7mol/L telmisartan +10~-7mol/L AngⅡ)and TP group(1mmol/L pyridoxamine +10~-7mol/L telmisartan +10~-7mol/L AngⅡ). The proliferation was accessed by methyl thiazolyl tetrazoliuin(MTT),and the migration was assessed by Scratch Wound Motility.The concentration of cellular supernatant advanced glycation end-products(AGEs) was masured by enzyme-linked immunosorbent assay(ELISA),and the levels of internal reactive oxygen species(ROS) were dectected by flow cytometry. Real-time fluorescence quantitative PCR was used to examine the expression of receptor for AGEs(RAGE),nuclear factor of kappaB P65(NF-κB-P65),nicotinamide adenine phosphate oxidase P47phox(NADPH P47phox) and matrix metalloproteinase-9 (MMP-9) mRNA,and Western Blot (WB) to the protein level of RAGE and NF-κB-P65.Results1.The MTT OD value and migrated cells in AngⅡgroup were significantly higher than that in control group(P<0.01).Compared with the AngⅡgroup,they were significantly lower in P,T and TP groups(P<0.01);and they were lower even more in TP group than that in P and T groups(P<0.01).2.Compared with the control group,the cellular supernatant AGEs concentration and the levels of intercellular reactive oxygen species(ROS) were significantly higher in AngⅡgroup(P<0.001).Compared with the AngⅡgroup,P group and T group after intervention significantly inhibited the production of AGEs and ROS, furthermore the tow drugs applied jointly maintained more significance than separately(P<0.001).3.The RAGE,NF-κB-P65,NADPH oxidaseP47phox and MMP-9 mRNA levels in AngⅡgroup all were significantly higer than that in control group(P<0.01).Compared with the AngⅡgroup,they were significantly decreased in P,T and TP group(sP<0.01);and they were decreased even more in TP group than that in P and T groups(P<0.01).4.The protein level of RAGE and NF-κB-P65w?as examined by Western Blot: Compared with the control group,it was significantly higer in AngⅡgroup,while drug intervention groups had the lower expression of them(P<0.01)than that in AngⅡgroup,furthermore TP group had even lower one than T and P groups(P<0.01).Conclusions1.Pyridoxamine can inhibit the VSMCs proliferation and migration induced by AngⅡ, and its mechanism is probably associated with it inhibiting the production of AGEs,reducing the intercellular ROS level and the expression of RAGE,NF-κB-P65,MMP-9 and NADPH P47phox.2.Telmisartan inhibits the VSMCs proliferation and migration induced by AngⅡ,via blocking AngⅡfrom binding to its AT1R,reducing the intercellular ROS level and inhibiting the expression of NF-κB-P65,MMP-9 and NADPH P47phox.3.Pyridoxamine and telmisartan can synergistically inhibit the VSMCs proliferation and migration induced by AngⅡ,its mechanism maybe that the two drugs employed together can synergistically reduce the intercellular ROS level and inhibit the expression of NF-κB-P65,MMP-9 and NADPH P47phox through the two aspects AGEs and AngⅡ.