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.

参考文献

[1]Clement ME, Hicks CB.Syphilis on the rise: what went wrong[J]. JAMA, 2016, 315:2281-2283.
[2]Lu P, Zheng DC, Fang C, et al. Cytokines in cerebrospinal fluid of neurosyphilis patients: identification of urokinase plasminogen activator using antibody microarrays[J]. J Neuroimmunol, 2016, 293:39-44.
[3]Radolf JD, Kumar S. The treponema pallidum outer membrane[J]. Curr Top Microbiol Immunol, 2018,415:1-38.
[4]Liu WN, Jiang XY, Ren J, et al. Tp47 induces cell death involving autophagy and mTOR in human microglial HMO6 cells[J].Int Immunopharmacol, 2019,74.doi: 10.1016/j.intimp.2019.04.013.
[5]Coureuil M, Jamet A, Bille E, et al.Molecular interac-tions between Neisseria meningitidis and its human host[J]. Cell Microbiol, 2019, 21:e13063.doi: 10.1111/cmi.13063.
[6]Dalal PJ, Muller WA, Sullivan DP. Endothelial cell calcium signaling during barrier function and inflammation[J]. Am J Pathol,2020, 190: 535-542.
[7]Lin SW, Gao ZX, Lin LR, et al. Treponema pallidum enhances human monocyte migration and invasion by dysregulating the MMP/TIMP balance[J]. Int Immunopharmacol, 2019, 75.doi: 10.1016/j.intimp.2019.105744.
[8]Radeva MY, Waschke J. Mind the gap: mechanisms regulating the endothelial barrier[J]. Acta Physiologica, 2017, 222.doi: 10.1111/apha.12860.
[9]Cong X, Kong W. Endothelial tight junctions and their regulatory signaling pathways in vascular homeostasis and disease[J]. Cell Signal, 2020, 66:109485.doi: 10.1016/j.cellsig.2019.109485.
[10]Greene C, Hanley N, Campbell M. Claudin-5: gate-keeper of neurological function[J]. Fluids Barriers CNS, 2019, 16:3. doi: 10.1186/s12987-019-0123-z.
[11]武勰,晏馥霞.紧密连接对肺通透性影响的研究进展[J].基础医学与临床,2021,41: 272-276.
[12]Yang XS, Liu MY, Zhang HM, et al. Protein kinase C-delta mediates sepsis-induced activation of complement 5a and urokinase-type plasminogen activator signaling in macrophages[J]. Inflamm Res, 2014, 63:581-589.
[13]Lai KC, Huang AC, Hsu SC, et al. Benzyl isothiocyanate (BITC) inhibits migration and invasion of human colon cancer HT29 cells by inhibiting matrix metalloproteinase-2/-9 and urokinase plasminogen (uPA) through PKC and MAPK signaling pathway[J]. J Agric Food Chem, 2010, 58:2935-2942.

[1]	陈笑天, 陈翀, 罗云萍. MAFB对肿瘤相关巨噬细胞极化及功能的影响[J]. 基础医学与临床, 2024, 44(7): 965-973.
[2]	宋博渊, 吴雪丽, 李雪媛, 王莉莎, 陈阳. 沙门氏菌感染巨噬细胞的动态转录物组分析[J]. 基础医学与临床, 2024, 44(6): 779-785.
[3]	张迪雅, 李云帆, 郭泓江, 仇佳星, 王钰铖, 鞠瑞, 郭磊. 胶质瘤条件培养基促进巨噬细胞糖酵解及表型转化[J]. 基础医学与临床, 2024, 44(5): 599-605.
[4]	赵东, 查世乾, 王易轩, 潘舟, 于文蓁, 胡克. 巨噬细胞向肌成纤维细胞转化促进LPS诱导的急性肺损伤模型小鼠肺纤维化[J]. 基础医学与临床, 2024, 44(3): 281-287.
[5]	余晗俏, 李超, 余育斌, 冯丽娜, 盛晓生, 叶晓霞, 王林燕. 抑制髓样细胞触发受体-1(TREM-1)减轻慢性间歇性低氧诱发的模型小鼠动脉粥样硬化[J]. 基础医学与临床, 2024, 44(3): 368-373.
[6]	陈卫卫, 廖煌, 史振鸿, 罗颖. FHL2通过NF-κB信号通路调节THP-1巨噬细胞泡沫化[J]. 基础医学与临床, 2024, 44(2): 204-209.
[7]	陈辛, 陈俊, 徐幸, 王旭. 组蛋白去甲基化酶抑制剂GSK-J4减轻高脂诱导的肥胖小鼠体质量[J]. 基础医学与临床, 2023, 43(9): 1353-1358.
[8]	杨雅倩, 潘艳芳, 屈梦扬, 方艳, 应小平, 张玫倩, 魏静. 外泌体在肝癌前病变发生发展中作用的研究进展[J]. 基础医学与临床, 2023, 43(9): 1453-1456.
[9]	张宝瑞, 马春晓, 秦历杰. STAT1诱导的LINC00987调节miR-223-3p/FZD4促进人胶质母细胞瘤相关巨噬细胞向M2型极化[J]. 基础医学与临床, 2023, 43(7): 1060-1068.
[10]	李晓娜, 齐先梅, 张田甜, 王婧. 沉默Fcgr3降低SiO₂诱导的小鼠肺泡巨噬细胞系MH-S炎性因子表达[J]. 基础医学与临床, 2023, 43(6): 904-908.
[11]	赵璐, 刘洋. γ-氨基丁酸促进水疱性口炎病毒感染小鼠巨噬细胞[J]. 基础医学与临床, 2023, 43(6): 936-940.
[12]	赵永晶, 仇佳星, 张迪雅, 郭泓江, 王钰铖, 鞠瑞, 郭磊. 稳定敲除Mcart-1下调RAW264.7巨噬细胞增殖能力和氧化磷酸化水平[J]. 基础医学与临床, 2023, 43(4): 568-575.
[13]	张凤云, 赵志浩, 张卓琦. miR-34a对高糖状态下小鼠巨噬细胞系RAW264.7极化及炎性因子分泌的影响[J]. 基础医学与临床, 2023, 43(12): 1808-1813.
[14]	张晨, 蒲翔, 苏进, 唐依莲, 曾宪法. 肺泡巨噬细胞诱导免疫应答对肺组织炎性损伤作用机制的研究进展[J]. 基础医学与临床, 2023, 43(10): 1585-1589.
[15]	杨雄, 靳潇潇, 何文强. 肾上皮细胞损伤与肾结石Randall斑块形成[J]. 基础医学与临床, 2022, 42(9): 1454-1458.

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

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价