牙龈卟啉单胞菌对人脐静脉血管内皮细胞cGMP生成的影响

Effect of Porphyromonas gingivalis on cGMP production in cultured human umbilical vein endothelial cells

目的:体外研究牙龈卟啉单胞菌(Porphyromonas gingivalis,Pg)对人脐静脉血管内皮细胞(human umbilical vein endothelial cell,HUVEC)cGMP水平的影响。方法:用Pg ATCC 33277分别以感染复数(multiplicity of infection,MOI)1∶10、1∶50、1∶250干预HUVEC,并以未受Pg ATCC 33277干预的HUVEC作为阴性对照,分别培养4、12、36 h,在倒置显微镜下观察各组细胞形态;125I cGMP放射免疫试剂盒检测各组HUVECcGMP的水平。结果:与对照组相比,Pg ATCC 33277分别以MOI 1∶10、1∶50、1∶250干预HUVEC 4、12、36 h后,实验各组HUVEC的细胞形态未见明显改变,仍呈典型的"铺路石"状单层贴壁生长;Pg ATCC 33277 MOI1∶250时可呈时间依赖性地降低HUVEC cGMP水平,而同一时间点内,各浓度组的cGMP水平并无明显差异。结论:短时间内,Pg ATCC 33277对HUVEC的细胞形态无明显影响,但可降低HUVEC cGMP的生成,HUVECNO生物利用度下降。

AIM： To observe the effect of Porphyromonas gingivalis ATCC 33277 （Pg ATCC 33277 ） on the production of cyclic guanosine monophosphate （cGMP） in cultured human umbilical vein endothelial cells （HUVEC）. METHODS： Pg ATCC 33277 was cultured anaerobically, and HUVEC were treated with Pg ATCC 33277 at multi- plicities of infection （MOI） of 1 ： 10, 1 ： 50 and 1 ： 250 for 4,12 and 36 h. HUVEC were observed using an inverted microscope and the intracellular cGMP levels in the cultured HUVEC were determined using radioimmunoassay. Detec- tion of eGMP was used as a reporter assay for the bioavailability of NO. RESULTS ： Compared with uninfected control group, HUVEC co-incubated with Pg ATCC 33277 for 4, 12 and 36 h at MOI of 1 ： 10, 1 ： 50 and 1 ： 250 respectively presented as an intact monolayer. Pg ATCC 33277 infection time-dependently reduced eGMP-production in HUVEC （P 〈 0.05 ）. Dose-dependence was not observed at all time points. CONCLUSION： Our results showed that within certain time period, the morphology of HUVEC is not affected with Pg. Pg. can inhibit cGMP production in human umbilical vein endothelial cells.