南方医科大学学报 ›› 2016, Vol. 36 ›› Issue (01): 73-.

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血小板活化因子受体在伴放线放线杆菌致病过程中的作用

王勤,轩东英,钟德钰,屈娅荣,余晶仪,曹虹,章锦才   

  • 出版日期:2016-01-20 发布日期:2016-01-20

Role of platelet-activating factor receptor in adhesion and invasion of Aggregatibacter
actinomycetemcomitans in human umbilical vein endothelial cells

  • Online:2016-01-20 Published:2016-01-20

摘要: 目的研究血小板活化因子受体(PAFR)在磷脂酰胆碱(PC)阳性伴放线放线杆菌(Aa)黏附侵袭人脐静脉血管内皮细胞
(HUVEC)过程中的作用。方法利用PAFR拮抗剂(CV3988)和抗PAFR抗体作用于HUVEC 30 min 后,观察PC阳性Aa 对
HUVEC黏附和侵袭的影响;并采用MTT法分析PC阳性和阴性Aa诱导细胞损伤的情况。结果采用100、200、500 nmol/L的
PAFR拮抗剂预处理HUVEC,显著减少了PC 阳性Aa 对HUVEC的黏附和侵袭(P<0.001),黏附率分别为对照组的(36.29±
3.52)%,(19.04±3.35)%和(7.69±3.19)%;侵袭率分别为对照组的(12.12±1.58)%,(7.08±0.29)%和(2.60±2.26)%。用抗PAFR抗
体预处理HUVEC后Aa对HUVEC的黏附率和侵袭率分别为(50.05±5.28)%和(39.09±6.50)%,显著降低了Aa对宿主细胞的黏
附和侵入(P<0.001)。采用200 nmol/L 和500 nmol/L 的PAFR拮抗剂和25 μg/mL抗PAFR抗体预处理HUVEC后,PC阳性的
Aa与细胞作用以后,细胞的存活率显著升高(P<0.001),从(25.39±9.33)%分别升高到(91.12±3.14)%,(94.12±2.15)%和(65.5±
1.87)%。而PC阴性的Aa菌株中,预处理组与未预处理的组相比,细胞活力并没有显著增加(P>0.05)。结论我们发现PAFR在
PC阳性的Aa对宿主细胞的粘附、侵袭及诱导胞死亡的过程中发挥了重要作用。

Abstract: Objective To explore the role of platelet-activating factor receptor (PAFR) in adhesion and invasion of phosphorylcholine
(PC)-positive Aggregatibacter actinomycetemcomitans in cultured human umbilical vein endothelial cells (HUVEC).
Methods Cultured HUVECs were pretreated with the PAFR antagonist CV3988 or anti-human PAFR monoclonal antibody for
30 min before infection with PC-positive or -negative A. actinomycetemcomitans strains. The bacterial adhesion and invasion and
cytotoxicity in the cells were examined using MTT assay. Results Pretreatment with PAFR antagonists at 100, 200 and 500
nmol/L significantly reduced the adhesion rate []36.29±3.52)%, (19.04±3.35)% and (7.69±3.19%), respectively] and invasion rate
[(12.12 ± 1.58)% , (7.08 ± 0.29)% and (2.60 ± 2.26)% , respectively] of PC-positive A.actinomycetemcomitans in HUVECs. Similarly,
pretreatment with anti-PAFR antibody also significantly reduced A.actinomycetemcomitans adhesion and invasion in HUVECs
[(50.05 ± 5.28)% and (39.09 ± 6.50)% , respectively]. Pretreatment with PAFR antagonist (200 and 500 nmol/L) and anti-PAFR
antibody (25 μg/mL) significantly increased the viability of HUVECs incubated with PC-positive A.actinomycetemcomitans from
(25.39 ± 9.33)% to (91.12 ± 3.14)% , (94.12 ± 2.15)% and (65.5 ± 1.87)% , respectively, but such pretreatments did not increase the
viability of cells incubated with PC-negative A.actinomycetemcomitans. Conclusions PAFR plays an important role in the
adhesion, invasion, and cytotoxicity of PC-positive A.actinomycetemcomitans in cultured HUVECs.