›› 2015, Vol. 35 ›› Issue (7): 525-531.

• Basic Research • Previous Articles     Next Articles

The effect ofⅠand Ⅳ fimA genotypes of porphyromonas gingivalis on the production of endothelin 1 and nitric oxide by co-cultured human umbilical vein endothelial cells with human artery smooth muscle cells

  

  • Received:2015-03-23 Revised:2015-05-06 Online:2015-07-28 Published:2015-08-04

Abstract: Abstract: Objective To observe the effects of different fimA genotypes of Porphyromonas gingivalis(P.gingivalis) on the production of endothelin1 (ET 1) and nitric oxide (NO) by co-cultured human umbilical vein endothelial cells (HUVECs) with human umbilical artery smooth muscle cells (HUASMCs).Methods P.gingivalis ATCC33277 (typeⅠfimA gene) and W83 (type ⅣfimA gene) were cultured anaerobically in standard condition,and a novel co-culture system of HUVECs and HUASMCs was treated with different fimA genotypes of P.gingivalis for 2,8,24 and 48h.At different time points,the supernatant was collected,the levels of ET 1 were determined by ELISA,and the levels of NO were determined by nitrate reductase.A negative control group (blank control) and a positive control group (1ug/mL E.coli LPS) were set in each experimental group.Results The co-culture system of HUVECs and HUASMCs produced ET 1 and NO with Ⅰand ⅣfimA genotypes of P.gingivalis stimulation.Compared with the negative and positive control groups,differences were observed concerning the ET 1 and NO production and ET 1/NO level in the experimental group.In terms of overall trend of the production of ET 1 and NO,the group ofⅠfimA genotype of P.gingivalis was similar to the negative control group,while the group of ⅣfimA genotype of P.gingivalis was similar to the positive control group.ⅣfimA genotype of P.gingivalis demonstrated more secretion of ET 1 and a lower amount of NO compared toⅠfimA genotype of P.gingivalis.At 48h,co-cultured HUVECs and HUASMCs infected by ⅣfimA genotype of P.gingivalis showed a significant imbalance of ET 1/NO.Conclusions Stimulated by different fimA genotypes of P.gingivalis,the production of ET 1 and NO by co-cultured HUVECs and HUASMCs,and the ET 1/NO level were significantly different between two fimA genotype of P.gingivalis,which may be related to the native virulence of the bacteria.ⅣfimA genotype of P.gingivalis could stimulate and influence co-cultured HUVECs and HUASMCs,and is more likely to cause endothelial dysfunction thanⅠfimA genotype P.gingivalis.

Key words: Porphyromonas gingivalis, HUVECs, HUASMCs, endothelin-1, nitric oxide

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