重组梅毒螺旋体蛋白Tp47通过诱导uPA增加血管内皮细胞通透性

doi:10.16352/j.issn.1001-6325.2022.01.018

基础医学与临床 ›› 2022, Vol. 42 ›› Issue (1): 120-125.doi: 10.16352/j.issn.1001-6325.2022.01.018

重组梅毒螺旋体蛋白Tp47通过诱导uPA增加血管内皮细胞通透性

于宝丹¹, 陈聪², 柯吴坚², 吕萍^2*

1.广州医科大学附属第一医院检验科, 广东广州 510120;
2.南方医科大学皮肤病医院皮肤科, 广东广州 510095

收稿日期:2021-03-23 修回日期:2021-06-03 出版日期:2022-01-05 发布日期:2022-01-05
通讯作者: ^* lvping75@163.com
基金资助:
广东省公益研究与能力建设资金(2017A020215152)

Recombinant Treponema pallidum protein Tp47 increases permeability of vascular endothelial cells through inducing uPA

YU Bao-dan¹, CHEN Cong², KE Wu-jian², LYU Ping^2*

1. Department of Laboratory Medicine, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou 510120;
2. Department of Dermatology, Dermatology Hospital, Southern Medical University, Guangzhou 510095,China

Received:2021-03-23 Revised:2021-06-03 Online:2022-01-05 Published:2022-01-05
Contact: ^* lvping75@163.com

摘要/Abstract

摘要： 目的探讨重组梅毒螺旋体蛋白Tp47(rTpp47)对单核-巨噬细胞系THP-1合成尿激酶型纤溶酶原激活物(uPA)的调控及其对人脐静脉/血管内皮细胞(HUVECs)通透性的影响。方法用rTpp47刺激THP-1细胞24 h后,分别收集细胞培养上清和THP-1细胞,用ELISA和Western blot检测THP-1细胞表达的uPA含量;用THP-1细胞培养上清刺激人单层血管内皮细胞后,使用FITC-葡聚糖评价单层内皮细胞通透性的变化,用Western blot检测uPA对HUVECs细胞紧密连接蛋白claudin-5表达的影响以及PKC信号通路是否参与rTpp47诱导的THP-1细胞表达uPA。结果重组蛋白rTpp47刺激THP-1细胞合成和分泌的uPA显著高于对照组(P＜0.05,P＜0.001);用rTpp47与THP-1细胞共培养24 h后收集的细胞培养上清刺激单层血管内皮细胞12和24 h,实验组血管内皮细胞相对通透性显著高于对照组(P＜0.05,P＜0.000 1);uPA活性抑制剂阿米洛利(amiloride)抑制了rTpp47刺激THP-1细胞分泌uPA(P＜0.000 1),改善了rTpp47对HUVECs的claudin-5蛋白表达的抑制作用(P＜0.05),抑制了rTpp47诱导的THP-1细胞表达蛋白激酶C(PKC)的磷酸化(P＜0.05)。结论重组蛋白rTpp47可能通过激活PKC信号通路刺激THP-1细胞合成和分泌的uPA,uPA作用于血管内皮细胞,可能导致血管内皮细胞通透性升高。

关键词: 梅毒螺旋体, 重组蛋白Tp47(rTpp47), 尿激酶型纤溶酶原激活物(uPA), 巨噬细胞, 人脐静脉/血管内皮细胞(HUVECs)

Abstract: Objective To investigate the effect of recombinant Treponema pallidum protein Tp47 (rTpp47) on the expression of uPA by THP-1 macrophages and on the permeability of human umbilical vein/vascular endothelial cells(HUVECs). Methods THP-1 cells were stimulated with rTpp47 for 24 hours and then the culture supernatant and THP-1 cells were collected respectively. The expression of urokinase-type plasminogen activator (uPA) in THP-1 cells was detected by ELISA and Western blot. After stimulation of HUVECs monolayers by THP-1 cell culture supernatant, FITC-dextran was used to evaluate the permeability of HUVECs. Western blot method was used to observe the effect of uPA on the expression of tight junction protein claudin-5 of HUVECs and the protein kinase C(PKC) signaling pathway of THP-1 cells stimulated by rTpp47. Results The synthesis and secretion of uPA of THP-1 cells stimulated by rTpp47 were significantly higher than those in the control group(P＜0.05, P＜0.001). When HUVECs were stimulated by the THP-1 cell supernatant for 12 hours and 24 hours, the relative permeability of vascular endothelial cells in the experimental group was significantly higher than the control group (P＜0.05 for 12 hours and P＜0.000 1 for 24 hours). Amiloride, the synthetic inhibitor of uPA, significantly inhibited the uPA expression and secretion of THP-1 cells stimulated by rTpp47 (P＜0.0001), alleviated the inhibition effect of rTpp47 on claudin-5 expression of HUVECs (P＜0.05), and significantly suppressed PKC phosphorylation(P＜0.05) in THP-1 cells induced by rTpp47. Conclusions The rTpp47 can stimulate the synthesis and secretion of uPA in THP-1 cells through a potential mechanism of activating PKC signaling pathway and uPA may increase the permeability of vascular endothelial cells.

Key words: Treponema pallidum, recombinant Tp protein Tp47(rTpp47), urokinase-type plasminogen activator(uPA), macrophages, human umbilical vein/vascular endothelial cells(HUVECs)

中图分类号:

于宝丹, 陈聪, 柯吴坚, 吕萍. 重组梅毒螺旋体蛋白Tp47通过诱导uPA增加血管内皮细胞通透性[J]. 基础医学与临床, 2022, 42(1): 120-125.

YU Bao-dan, CHEN Cong, KE Wu-jian, LYU Ping. Recombinant Treponema pallidum protein Tp47 increases permeability of vascular endothelial cells through inducing uPA[J]. Basic & Clinical Medicine, 2022, 42(1): 120-125.

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重组梅毒螺旋体蛋白Tp47通过诱导uPA增加血管内皮细胞通透性

Recombinant Treponema pallidum protein Tp47 increases permeability of vascular endothelial cells through inducing uPA

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