Effects Of New Pyrazole Derivatives On Apoptosis Of Vascular Endothelial Cells And A549 Lung Cancer Cells And The Corresponding Mechanisms

Effects of new pyrazole derivatives on apoptosis of vascular endothelial cells and A549 lung cancer cells and the corresponding mechanismsVascular endothelial cells(VEC)form the inner lining of all blood vessels and function to maintain vascular tone and anticoagulant properties of blood vessels.VEC apoptosis plays important roles in many diseases,including atherosclerosis and cancer. Previously,we found that deprivation of serum and FGF-2 could induce HUVEC apoptosis.In this study,in the light of chemical genetics,we used new pyrazole derivatives as tools to study the molecular mechanism of apoptosis in HUVECs and A549 lung cancer cells.METHODS:1.HUVECs were gained as described previously[Jaffe et al.,1973].2.The morphological changes were observed under a phase contrast microscope.3.The cell proliferation was measured by MTT assay.4.DNA nuclear fragmentation was analyzed by acridine orange staining.5.The TdT-mediated dUTP nick end labeling technique was used to detect in situ nuclear DNA fragmentation and count apoptosis ratio.6.The expressions and distributions of integrinβ4 and p53 were analysed by immunofluorescence assay.7.The fluorescence probe,DCHF was used to analyze ROS level.RESULTS:1 When we treated HUVECs with Ethyl 3-(o-chlorophenyl)-5-methyl-1H-pyrazole-4-carboxylate(4a),Ethyl 5-methyl-3-(o-nitrophenyl)-1H-pyrazole-4-carboxylate(4b)at 100μM, respectively,the viability of HUVECs decreased obviously.But,when we treated HUVECs with 1-Phenyl-3-p-tolyl-1H-pyrazol-5-ol(4f)at 25μM,the viability of HUVECs increased obviously.2 Ethyl 3-(o-chlorophenyl)-5-methyl-1-phenyl-1H-pyrazole-4-carboxylate(4d, MPD)(25μM)inhibited the apoptosis induced by deprivation of serum and FGF-2,but promoted the apoptosis at 100μM.3 The high levels of integrinβ4 and p53 induced by the deprivation of serum and FGF-2 could be depressed by the treatment of MPD(25μM).4 MPD(25μM)could depress the accumulation of intracellular ROS.5 MPD(25,50,100μM)could not affect the morphology and viability of HUVECs in the present of serum and FGF-2.6 When treated with 4a(100μM)and 4d(MPD)(50,100μM),respectively,A549 cell growth was obviously suppressed(p＜0.01)and the cells were induced to apoptosis.7 4b,3-(o-nitrophenyl)-1-phenyl-1H-pyrazol-5-ol(4e),4f,had no obvious effects on A549 cell growth.CONCLUSION:1 In the absence of serum and FGF-2,4f(25μM)could increase the viability of HUVECs obviously,but 4a and 4b could depress the viability of HUVECs obviously at 100μM,respectively..2 MPD inhibited the apoptosis of HUVECs induced by the deprivation of serum and FGF-2 at low concentration(25μM),but promoted apoptosis at high concentrations(50,100μM).3 MPD(25μM)inhibited the apoptosis through down-regulating the levels of integrinβ4,p53,and ROS.4 MPD(25,50,100μM)could not affect the viability of HUVECs in the present of serum and FGF-2.5 The growth of A549 cells decreased after treatment with 4a at 100μM for 24 h or 48 h,respectively.6 The growth of A549 cells decreased after treatment with MPD 50,100μM, respectively.