Study on the mechanism of PALM3 expression on human umbilical vein endothelial cells induced by Pseudomonas aeruginosa

doi:10.3877/cma.j.issn.1674-6902.2025.05.003

Abstract

Abstract:

Objective

To explore the mechanism of PALM3 on vascular endothelial cell injury induced by Pseudomonas aeruginosa (PA).

Methods

Human umbilical vein endothelial cells (HUVEC) were randomly divided into the control group, the control+ PA group, the siRNA-control+ PA group, and the siRNA-PALM3+ PA group. Control group: HUVEC cells + normal saline; Control+ PA group: HUVEC cells exposed to fluorescently labeled 5×10⁸ CFU/ml PA bacterial solution .The siRNA-Control+ PA group and the siRNA-PALM3+ PA group transfected cells with siRNA-Control and siRNA-PALM3 plasmids respectively, and PA was added 48 hours after transfection. It was detected that vascular endothelial fibronectin, vascular endothelial growth factor c (VEGF-C), and angiopoietin-2 (ANGPT2) in the cell supernatant; The expressions of PALM3 and nuclear factor NF-κB in cells were detected. The cell survival rate was compared by MTT, and cell apoptosis was analyzed by flow cytometry.

Result

Compared with the Control group, the expression of PALM3 protein in the cells of the Control+ PA group and the siRNA-Control+ PA group increased (P<0.05); Compared with the Control+ PA group and the siRNA-Control+ PA group, the expression level of PALM3 protein in the cells of the siRNA-PALM3+ PA group decreased, and the expression level of NF-κB protein decreased. The fibronectin (235.7±17.5), VEGF-C(0.72±0.05), and ANGPT2 (807.8±24.6) in the siRNA-PALM3+ PA group were higher than those in the Control+ PA group (109.9±5.8) and VEGF-C(0.49±0.06), ANGPT2 (464.16±26.27), and fibronectin (188.9±8.6), VEGF-C (0.49±0.02), ANGPT2 (510.57±27.6) in the siRNA-Control+ PA group (P<0.05); The cell survival rate of the siRNA-PALM3+ PA group (1.102±0.125) was higher than that of the Control+ PA group (0.870±0.052) and the siRNA-Control+ PA group (0.887±0.054) (P<0.05); The cell survival rates of the Control+ PA group and the siRNA-Control+ PA group after 6 hours were (2.760 ± 0.744) and (2.777 ± 0.604), respectively, which were higher than that of the Control group (1.662±0.150). Compared with the Control+ PA group and the siRNA-Control+ PA group, the cell survival rate in the siRNA-PALM3+ PA group (3.195±1.342) increased.

Conclusion

Down-regulation of PALM3 has a protective effect on the inflammatory response of pulmonary vascular endothelial cell injury caused by Pseudomonas aeruginosa, and is negatively correlated with vascular permeability.

Key words: Acute lung injury, PALM3, Pseudomonas aeruginosa, Vascular endothelial cells, Proinflammatory cytokines

Xiulin Wu, Liang Guo, Jun Cai, Zhiqiang He, Yue Wang, Tianyi Yang, Min Tang, Mingxia Li, Zhiyong Yang, Bin Yi, Wei Xiong, Jiangrong Liao, Xueling Wu. Study on the mechanism of PALM3 expression on human umbilical vein endothelial cells induced by Pseudomonas aeruginosa[J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(05): 679-684.

/ / Recommend

Add to citation manager EndNote|Ris|BibTeX

URL: https://zhfbjbzz.cma-cmc.com.cn/EN/10.3877/cma.j.issn.1674-6902.2025.05.003

https://zhfbjbzz.cma-cmc.com.cn/EN/Y2025/V18/I05/679

Figures/Tables 4

References 32

1	马李杰，李王平，金发光，等. 急性肺损伤/急性呼吸窘迫综合征发病机制的研究进展[J/OL]. 中华肺部疾病杂志(电子版), 2013, 6(1): 65-68.
2	张鹏，史慢慢，马辉，等. 急性呼吸窘迫综合征患者机械通气动脉血二氧化碳分压变异率与预后风险相关性[J/OL]. 中华肺部疾病杂志(电子版), 2025, 18(2): 226-230.
3	Fatma Yıldırım, Írem Karaman, Akın Kaya. Current situation in ARDS in the light of recent studies: Classification, epidemiology and pharmacotherapeutics[J]. Tuberk Toraks, 2021, 69(4): 535-546.
4	Huang X, Xiu H, Zhang S, et al. The role of macrophages in the pathogenesis of ALI/ARDS[J]. Mediators Inflamm, 2018, 2018: 1264913.
5	ZhouJ, Peng Z, Wang J. Trelagliptin alleviates lipopolysaccharide (LPS)-induced inflammation and oxidative stress in acute lung injury mice[J]. Inflammation, 2021, 44(4): 1507-1517.
6	Huikang Xu, Shiying Sheng, Weiwei Luo, et al. Acute respiratory distress syndrome heterogeneity and the septic ARDS subgroup[J]. Front Immunol, 2023, 14: 1277161.
7	Cao J, Liu M, Feng S, et al. Glaucocalyxin A alleviates lipo-polysaccharide-induced inflammation and apoptosis in pulmo-nary microvascular endothelial cells and permeability injury by inhibiting STAT3 signaling[J]. Exp Ther Med, 2022, 23(4): 313.
8	Matthay MA, Zemans RL, Zimmerman GA, et al. Acute respiratory distress syndrome[J]. Nat Rev Dis Primers, 2019, 5(1): 18.
9	Máca J, Jor O, Holub M, et al. Past and present ARDS mortality rates: a systematic review[J]. Respir Care, 2017, 62(1): 113-22.
10	Haute GV, Luft C, Antunes GL, et al. Anti-inflammatory effect of octyl gallate in alveolar macrophages cells and mice with acute lung injury[J]. J Cell Physiol, 2020, 235(9): 6073-6084.
11	Wei Wang, Shengtian Mu, Dongli Yan, et al. Comprehending toll-like receptors: pivotal element in the pathogenesis of sepsis and its complications[J]. Front Immunol, 2025, 16: 1591011.
12	Hellmann J, Tang Y, Zhang MJ, et al. Atf3 negatively regulates Ptgs2/Cox2 expression during acute inflammation[J]. Prostaglandins Other Lipid Mediat, 2015, 116-117: 49-56.
13	Wang Z, Xu D, Ding HF, et al. Loss of ATF3 promotes Akt activation and prostate cancer development in a Pten knockout mouse model[J]. Oncogene, 2015, 34(38): 4975-4984.
14	Pu H, Wang X, Su L, et al. Heroin activates ATF3 and CytC via c-Jun N-terminal kinase pathways to mediate neuronal apoptosis[J]. Med Sci Monit Basic Res, 2015, 21: 53-62.
15	Zhang C, Wu X, Zhao Y, et al. SIGIRR inhibits toll-like receptor 4，5，9-mediated immune responses in human air-way epithelial cells[J]. Mol Biol Rep, 2011, 38(1): 601-609.
16	Bodaszewska-Lubas M, Liao Y, Zegar A, et al. Dominant-negative form of SIGIRR: SIGIRRΔE8 promotes tumor growth through regulation of metabolic pathways[J]. J Interferon Cytokine Res, 2022, 42(9): 482-492.
17	Chen X, Zhao Y, Wu X, et al. Enhanced expression of sin-gle immunoglobulin IL-1 recepter-related molecule amel-iorates Lipopolysaccharide-induced acute lung injury in mice[J]. Shock, 2011, 35(2): 198-204.
18	陈旭昕，孟激光，韩志海，等. 干扰paralemmin-3表达抑制脂多糖诱导的肺泡上皮细胞的炎性反应[J]. 西部医学，2015, 27(7): 964-966, 970.
19	Li S, Guo L, Zhao Y, et al. Silencing of Paralemmin-3 Protects Mice from lipopolysaccharide-induced acute lung injury[J]. Peptides, 2016, 76: 65-72.
20	Chen X, Wu X, Zhao Y, et al. A novel binding protein of sin-gle imnmnoglobulin IL-1 receptor-related nmlecule: Paralenmfin -3[J]. Biochem Biophys Res Commun, 2011, 404(4): 1029-1033.
21	Mougeot JC, Beckman MF, Hovan AJ, et al. Identification of single nucleotide polymorphisms (SNPs) associated with chronic graft-versus-host disease in patients undergoing allogeneic hematopoietic cell transplantation[J]. Support Care Cancer, 2023, 31(10): 587.
22	陈旭昕，孟激光，刘振千，等. 下调Paralemmin-3表达对脂多糖诱导的肺泡上皮细胞核转录因子-κB活性的影响[J]. 广东医学，2014, 35(19): 2969-2971.
23	Chen X, Tang L, Feng J, et al. Downregulation of paralemmin-3 ameliorates Lipopolysaccharide-induced acute lung injury in rats by regulating inflammatory response and inhibiting forma-tion of TLR4/MyD88 and TLR4 /TRIF complexes[J]. Inflammation, 2017, 40(6): 1983-1999.
24	Chen XX, Tang L, Fu YM, et al. Paralemmin-3 contributes to lipopolysaccharide-induced inflammatory response and is involved in lipopolysaccharide-Toll-like receptor-4 signaling in alveolar macrophages[J]. Int J Mol Med, 2017, 40(6): 1921-1931.
25	陈旭昕，孟激光，马白鸽，等. 姜黄素对脂多糖诱导的巨噬细胞炎症与氧化应激的影响及机制[J]. 广东医学，2018, 39(18): 2713-2717.
26	Cornish JA, Kloc M, Decker GL, et al. Xlcaax-1 is localized to the basolateral membrane of kidney tubule and other polarized epithelia during xenopus development[J]. Dev Biol, 1992, 150: 108-120.
27	Xu S, Pan X, Mao L, et al. Phospho-Tyr705 of STAT3 is a therapeutic target for sepsis through regulating inflammation and coagulation[J]. Cell Commun Signal, 2020, 18(1): 104.
28	Yunfeng Zhao, Xiulin Wu, Lanlan Qian, et al. Activating transcription factor 3 protects mice against pseudomonas aeruginosa-induced acute lung injury by interacting with lipopolysaccharide binding protein[J]. Mol Immunol, 2017, 90: 27-32.
29	Xuxin Chen, Lu Tang, Jian Feng, et al. Downregulation of paralemmin-3 ameliorates lipopolysaccharide-induced acute lung injury in rats by regulating inflammatory response and inhibiting formation of TLR4/MyD88 and TLR4/TRIF complexes[J]. Inflammation, 2017, 40(6): 1983-1999.
30	Zhaosheng Jin, Ka Chun Suen, Daqing Ma. Perioperative "remote" acute lung injury: recent update[J]. J Biomed Res, 2017, 31(2): 197-212.
31	Singer Mervyn, Deutschman Clifford S, Seymour Christopher Warren, et al. The third International consensus definitions for sepsis and septic shock (Sepsis-3)[J]. JAMA, 2016, 315(8): 801-810.
32	Di A, Mehta D, Malik AB. ROS-activated calcium signaling mechanisms regulating endothelial barrier function[J].Cell Calcium, 2016, 60(3): 2071-2075.

组　别	纤维连接蛋白	VEGF-C	ANGPT2
Control	125.6±4.33	0.28±0.03	570.3±29.4
Control＋PA	109.9±5.8^a	0.49±0.06^a	464.16±26.27^a
SiRNA-Control＋PA	188.9±8.6^ab	0.49±0.02^a	510.57±27.6^ab
SiRNA-Plam3＋PA	235.7±17.5^abc	0.72±0.05^abc	807.8±24.6^abc
F值	124.7	53.66	127.6
P值	0.0001	0.0001	0.0001

组　别	纤维连接蛋白	VEGF-C	ANGPT2
Control	125.6±4.33	0.28±0.03	570.3±29.4
Control＋PA	109.9±5.8^a	0.49±0.06^a	464.16±26.27^a
SiRNA-Control＋PA	188.9±8.6^ab	0.49±0.02^a	510.57±27.6^ab
SiRNA-Plam3＋PA	235.7±17.5^abc	0.72±0.05^abc	807.8±24.6^abc
F值	124.7	53.66	127.6
P值	0.0001	0.0001	0.0001

[1]	Lulu Li, Lihong Ma, Jiajia Jin, Wei Gu. Stimulator of interferon genes affects acute lung injury repair in mice through pulmonary macrophage efferocytosis [J]. Chinese Journal of Critical Care Medicine(Electronic Edition), 2024, 17(02): 97-103.
[2]	Xiaobo Zhang, Te Ba, Ruijuan Huang, Hongyu Wang. Research progress on the mechanism of mesenchymal stem cell-derived exosomes on improving acute lung injury [J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2025, 20(01): 81-85.
[3]	Tuo Shen, Feng Zhu. Clinical significance of heteroresistance in nonfermenting gram-negative bacilli infection in burn patients [J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2025, 20(01): 75-80.
[4]	Yanhui Jia, Yixuan Yuan, Hao Guan, Dahai Hu. Mechanistic analyses on the role of senescent cell clearance in alleviating acute lung injury in septic mice [J]. Chinese Journal of Injury Repair and Wound Healing(Electronic Edition), 2025, 20(01): 55-60.
[5]	Yi Tan, Xincai Xu, Qiufeng Wan, Bowei Cao, Chunxing Li, Yangchao Guo. Protective effects of ophiopogonin D on acute lung injury and intestinal mucosal barrier injury [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(04): 527-533.
[6]	Cailin Zhao, Qing Xiang, Hang Qian, Wen Shi, Lingxiao Qiu, Bin Wang. Identification of ferroptosis-related hub genes and immune subtypes in acute lung injury/acute respiratory distress syndrome based on bioinformatics analysis [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(04): 503-509.
[7]	Chun Yang, Yangwei You, Yu Zhou, Zechao Lin, Liang Liu, Kun Yuan. Study on the effects of interleukin-10 in mice with transfusion-related acute lung injury by regulating the helper T cell 17/regulatory T cell [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(03): 355-361.
[8]	Pengsen Wang, Huifeng Wu, Jianfang Wen, Yang Wei, Longhao Dong, Boqiang Liu, Zhan Li, Feng Shichun, Xiaodong Lei, Xiongxiong Wu. Expression of serum miR-146a in sepsis-associated acute lung injury and its prognostic relevance [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(02): 241-245.
[9]	Xiaolong Zong, Yuanxi Lin, Tianyi Zhang, Yaru Liu, Duanyang Li, Zhenyu Li. Pterostilbene attenuates acute lung injury in septic mice by inhibiting inflammatory responses and NETs formation [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2025, 18(01): 29-35.
[10]	Minlong Zhang, Cuiping Yang, Bo Wang, Yunjie Cui, Faguang Jin. miR-200b-3p suppressed the edema in seawater aspiration-induced ALI via inhibition the expression of HIF-1α [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(05): 696-700.
[11]	Yujuan Li, Fang Ai, Huanqing Xiong, Jian Chen, Gang Liu, Zhichao Li, Faguang Jin. Therapeutic effect of "Tanshinone ⅡA+ Osthole" formula on acute lung injury in mice [J]. Chinese Journal of Lung Diseases(Electronic Edition), 2024, 17(02): 171-177.
[12]	Ling Lin, Jingru Li, Ruihua Shen, Hui Lin, Xi Qiao. Bioinformatics analysis of hub genes of acute kidney injury and acute lung injury in mice [J]. Chinese Journal of Kidney Disease Investigation(Electronic Edition), 2024, 13(03): 134-144.
[13]	Haonan Li, Yupeng Zhang, Yan Fu, Jiwei Feng, Kai Liu, Wenkai Zhang. Advances in the study of Connexin 43 in lung diseases [J]. Chinese Journal of Critical Care & Intensive Care Medicine(Electronic Edition), 2024, 10(01): 60-65.
[14]	Wan Liu, Tingting Zhao, Minghui Wei. Pseudomonas aeruginosa injection for treatment of refractory high-output chyle leakage after cervical lymph node dissection: report of two cases with a literature review [J]. Chinese Journal of Clinicians(Electronic Edition), 2024, 18(10): 926-930.
[15]	Ke Yang, Zihe Cheng, Shengyu Wang. Research progress on application of multi-omics technologies in acute lung injury [J]. Chinese Journal of Diagnostics(Electronic Edition), 2025, 13(03): 159-164.

Please choose a citation manager

Content to export