Amlodipine Down-regulates PDGF-B Expression In HUVEC-12 Endothelial Cells Through NF-κB Pathway

PartⅠ. Change of PDGF-B Expression in HUVEC-12 Endothelial Cells Induced by Ox-LDLAIM To observe the mRNA and protein expression of PDGF-B in HUVEC-12 endothelial cells induced by oxidized low density lipoprotein(ox-LDL)of different concentration and different time.METHODS HUVEC-12 endothelial cells were treated with different concentration (0,25,50,100 mg/L)of ox-LDL for 24 hours. RT-polymerase chain reaction(RT-PCR)and Western blot analysis were used to detect the change of mRNA and protein expression of PDGF-B. Then 50 mg/L ox-LDL was chosen into HUVEC-12 cells for different time(0,3,6,12,24h). RT-PCR and Western blot analysis were also used to detect the change of mRNA and protein expression of PDGF-B respectively.RESULTS With the increasing of the concentration of ox-LDL, both the mRNA and protein expression of PDGF-B in the cells were significantly increased in the concentration of 50 mg/L but decreased in the concentration of 100 mg/L, so 50mg/L ox-LDL was treated with the cells for different time. With the increasing of the time of ox-LDL, the expression of PDGF-B was significantly increased in a time-dependent manner. P< 0.05,there was significant discrepancy. Integrated with documents, we chose the cells to be treated with 50mg/L ox-LDL for 24h.CONCLUSION Ox-LDL can affect the expression of PDGF-B in the HUVEC-12 endothelial cells.PartⅡ. Function of Amlodipine on the Expression of PDGF-B and NF-κB in HUVEC-12 Endothelial Cells Induced by Ox-LDLAIM To observe the effect of Amlodipine of different concentration on the Expression of PDGF-B and NF-κB in HUVEC-12 Endothelial cells induced by ox-LDL.METHODS Considering the result above-mentioned,we chose the concentration and time of ox-LDL treated to be 50 mg/L and 24h. The experiment groups included 5 groups:control group, ox-LDL group, different concentration(0.1,1.0,10.0μM)of Amlodipine pretreated for 0.5h and treated with 50 mg/L ox-LDL for 24h group. RT-PCR and Western-blot analysis were used to detect the mRNA expression of PDGF-B and the protein expression of PDGF-B and NF-κB.RESULTS RT-PCR and Western-blot analysis showed that with ox-LDL and Amlodipine treatment, the mRNA expression of PDGF-B and the protein expression of PDGF-B and NF-κB were greatly decreased in a concentration-dependent manner.CONCLUSION Amlodipine can down-regulate the expression of PDGF-B and NF-κB. PartⅢ. Function of Amlodipine and Inhibitor of NF-κB PDTC on the Expression of PDGF-B and NF-κB in HUVEC-12 Endothelial Cells Induced by Ox-LDLAIM To observe the effect of Amlodipine and inhibitor of NF-κB PDTC on the Expression of PDGF-B and NF-κB in HUVEC-12 Endothelial cells induced by ox-LDL to explore the role of Amlodipine on PDGF-B through NF-κB.METHODS Considering the result above-mentioned,we chose the concentration of Amlodipine treated to be 10.0μM. The experiment groups included 6 groups:control group, ox-LDL group, 10.0μM amlodipine and 20μM PDTC for 30 min respectively then incubated with ox-LDL at the same concentration for 24h group and treated with Amlodipine or PDTC alone without ox-LDL group. RT-PCR and ELISA analysis were used to detect the mRNA and the protein expression of PDGF-B, while Western-blot analysis and immunocytochemistry were used to detect the protein expression of NF-κB.RESULTS Amlodipine can down-regulate the expression of PDGF-B and NF-κB as effectively as the inhibitor of NF-κB named PDTC. CONCLUSION Amlodipine can down-regulate the expression of PDGF-B through NF-κB.