Influence of angiotensin-converting enzyme inhibitory peptide on endothelial cell proliferation and endothelin expression in human umbilical vein cells
- VernacularTitle:降血压肽对人脐静脉内皮细胞增殖及内皮素表达的影响
- Author:
Dong LIU
;
Lijun ZHANG
;
Shimin LI
;
Fang LIU
;
Shizhong LIANG
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2006;10(25):160-163
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND: As a kind of polypeptide, angiotensin-converting enzyme (ACE) inhibitory peptide can lower the blood pressure of human body through restraining the formation of angiotensin Ⅱ.OBJECTIVE: To investigate the influence of AGE inhibitory peptide on endothelial cell proliferation and endothelin expression in cultured human umbilical vein cells based on cellular and molecular levels in order to provide the experimental evidences for ACE inhibitory peptide to be the potential blood pressure-lowering health food.DESIGN: Repeated measures design.SETTING: School of Bioscience and Bioengineering, South China University of Technology; School of Applied Chemistry and Biological Technology,Shenzhen Polytechnic.MATERIALS: The experiment was carried out in the Institute of Applied Chemistry and Biological Technology, Shenzhen Polytechnic from September 2004 to March 2005. The AGE inhibitory peptide was provided by the Institute of Applied Chemistry and Biological Technology, Shenzhen Polytechnic. Under certain circumstance, 15 μ mol/L of the inhibitor was needed to decrease half of the AGE activity. The human umbilical vein endothelial cells of the 4th generation were cultured randomly in 7 groups with different concentrations: medium group, 150, 300 and 600 mg/L ACE inhibitory peptide groups, captopril group, norepinephrine(NE) group, and ACE inhibitory peptide+NE group.METHODS: ①The endothelial cells were cultured as recommended. The medium was M199+FBS(0.15, v/v)+penicillin(10 000 U/mL)+streptomycin (100 mg/L). After cellular fusion, the cells were carried on the passage with the ratio of 1:2. The 4th generation cells were used for experiment. ②M199(0.15, v/v) was contained in each group. ACE inhibitory peptides were added to make the final concentration 150, 300 and 600 mg/L in the 150, 300 and 600 mg/L ACE inhibitory peptide groups respectively. Captopril was added to make the final concentration 10-5 mol/L in the captopril group. NE was added to make the final concentration 100 μg/L in the NE group. ACE inhibitory peptide and NE were added to make the final concentration 300 mg/L and 100 μg/L in the ACE inhibitory peptide+NE group respectively. ③The state of cell growth was determined with cytometry. The contents of endothelial cells in the medium with different culture times were determined with radioimmunoassay. The expression of endothe lin mRNA was determined with reverse transcriptase-polymerase chain reaction. The expression of cellular endothelin protein was determined with immunohistochemical method.MAIN OUTCOME MEASURES: ①The influence of ACE inhibitory peptide on endothelial cell proliferation. ②The influence of ACE inhibitory peptide on the endothelin mRNA and endothelin protein.RESULTS: ①The influence of ACE inhibitory peptide on endothelial cell proliferation and endothelin secretion: Compared with the medium group,in the captopril and 150, 300, 600 mg/L ACE inhibitory peptide groups,the growth of endothelial cells was restrained and the endothelin content in the medium was lowered(P ＜ 0.01 or 0.05). NE could promote the growth of endothelial cells and the secretion of endothelin, but the cell density and endothelin content after treatment with ACE inhibitory peptide were similar to those in the medium group (P ＞ 0.05). ②The influence of ACE inhibitory peptide on the expressions of endothelin mRNA and protein in endothelial cells: Compared with the medium group, the expressions of endothelin mRNA and protein might be lowered in the captopril and 150,300, 600 mg/L ACE inhibitory peptide groups(P ＜ 0.01 or 0.05). The expressions of endothelin mRNA and protein could be up-regulated by NE.The gene expression after treatment with ACE inhibitory peptide was similar to that in the medium group(P ＞ 0.05).CONCLUSION: The ACE inhibitory peptides of different dosages can all restrain the growth of endothelial cells, lower the endothelin content, decrease the expression of endothelin gene and resist NE improved growth and secretion of endothelial cells in umbilical vein cell effectively.
