Ⅰ、ⅣfimA型牙龈卟啉单胞菌对内皮细胞与平滑肌细胞共培养体系产生内皮素-1及一氧化氮的影响

摘要/Abstract

摘要： ［摘要］目的研究不同fimA基因型牙龈卟啉单胞菌（Porphyromonas gingivalis,P.gingivalis）刺激人脐静脉血管内皮细胞（human umbilical vein endothelial cells,HUVECs）及人脐动脉血管平滑肌细胞（human umbilical artery smooth muscle cells，HUASMCs）共培养体系产生内皮素-1（endothelin-1，ET-1）和一氧化氮（NO）的水平。方法用ⅠfimA型P.gingivalis（ATCC 33277）和ⅣfimA型P.gingivalis（W83）分别刺激HUVECs-HUASMCs共培养体系，于2、8、24、48 h时收集细胞培养上清液，酶联免疫反应检测ET-1的含量，硝酸还原酶法测定NO的含量。各组均设阴性对照组（纯培养基）及阳性对照组（1 μg/mL E.coli-LPS）。结果 HUVECs-HUASMCs共培养体系在Ⅰ、ⅣfimA型P.gingivalis刺激作用下产生ET-1和NO的量以及ET-1/NO水平，与阴性及阳性对照组比较存在差异。ⅠfimA型P.gingivalis刺激细胞共培养模型分泌ET-1和NO情况的总趋势与阴性对照组相似、而ⅣfimA型P.gingivalis的刺激作用总趋势则与阳性对照组相似，ⅣfimA型P.gingivalis较ⅠfimA型P.gingivalis刺激共培养细胞可分泌更多的ET-1、而NO量减少，ⅣfimA型P.gingivalis感染48 h的细胞共培养模型表现出明显的ET-1/NO的失衡。结论不同fimA型P.gingivalis刺激共培养模型后产生ET-1及NO的情况及ET-1/NO的水平有明显差异，可能与其本身毒力相关，ⅣfimA型P.gingivalis比ⅠfimA型P.gingivalis更易引起内皮功能紊乱。

关键词: 牙龈卟啉单胞菌, 人脐静脉血管内皮细胞, 人脐动脉血管平滑肌细胞, 内皮素-1, 一氧化氮

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

中图分类号:

R780.2

李文军贾惠杰葛颂. Ⅰ、ⅣfimA型牙龈卟啉单胞菌对内皮细胞与平滑肌细胞共培养体系产生内皮素-1及一氧化氮的影响[J]. 口腔医学, 2015, 35(7): 525-531.

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