| Objectives: To determine the effect of fine particulate matter (PM2.5) on the viability of human umbilical vein endothelial cells (HUVEC) and the expression of angiotensin-converting enzyme (ACE) and angiotensin II type1receptor (AT1R) genes.Methords: Water-soluble and non water-soluble PM2.5samples were collected from non-industrial area in Changsha city between October2010and December2011. HUVEC cells were treated with various PM2.5(100ug/ml,200ug/ml and400ug/ml defined as low dose group, intermediate dose group and high dose group respectively). After twenty-four hours, Cell viability was checked by using MTT assay, Real time PCR and Western Blot were used to analyze the mRNA and protein expression of ACE and AT1R genes, respectively. To evaluate the time-dependent effect of PM2.5, HUVEC cells were treated with high dose of PM2.5for0h,12h,24h and48h, at each time point the experiments, Cell viability and expression of ACE and AT1R genes were determined. To evaluate the effect of perindopril on PM2.5intervention of HUVEC cells, Cells were co-administrated with perindopril (10umol/L) for twenty-four hours, at the end of the experiment, ACE and AT1R genes expression were determined.Results:1. The cell viability of HUVEC cells were inhibited by PM2.5 The Cell viability in low, intermediate and dose group were significantly higher than control group, this effect was dose-dependent (P<0.05). When exposed to the same agent as in high and intermediate dose, the group of non-water soluble components of the cell viability was lower than the water-soluble components group (P<0.05). The Cell viability in12h,24h and48h groups were significantly higher than Oh group, this effect was time-dependent (P<0.05).2. PM2.5affected the mRNA and protein expression level of ACE gene: intermediate, high dose of water-soluble component of PM2.5and various dose of non water-soluble in the mRNA and protein expression level of ACE were higher than the control group (P<0.05), and the ACE expression level of the high dose group was the highest. When exposed to the same agent as in high dose, the group of non-water soluble components of the ACE mRNA and protein expression level was higher than the water-soluble (P<0.05). Water-soluble components of the24h,48h groups and non-water soluble components of the various time groups of the ACE gene expression level was higher than0h group (P<0.05), and the ACE expression level of the24h group was the highest.3. PM2.5affected the mRNA and protein expression level of AT1R gene: high dose of water-soluble and non water-soluble component of PM2.5in the mRNA and protein of AT1R expression level were higher than the control group (P<0.05), when HUVEC cells were exposed to non water-soluble component of PM2.5the mRNA and protein expression level of ATlR gene was dose-dependent (P<0.05). When exposed to the same agent as in high dose, the group of non-water soluble components of the ATlR mRNA and protein expression level was higher than the water-soluble (P<0.05). The ATlR mRNA and protein expression level in12h,24h and48h groups were significantly higher than0h group, and the48h group was the highest (P<0.05).4. The mRNA and protein expression level of ACE, ATlR genes of PM2.5influence after intervention in perindopril: low, intermediate, high dose of water-soluble and non water-soluble component of PM2.5in the mRNA and protein of ACE expression level were higher than the non plus drug group (P<0.05). however, the ATIR mRNA and protein expression level showed no statistical difference (P>0.05).Conclusions:1. PM2.5could reduce HUVEC cells vitality in a dose and time dependent, especially the non water-soluble component of PM2.5.2. Both the water-soluble and non water-soluble component of PM2.5could increase the expression of ACE and ATIR genes in HUVEC cells,and the effect was more significantly after non water-insoluble PM2.5components induction.ACEI could inhibit the upregulation of the ACE gene induced by the PM2.5in HUVEC cells. |
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