硫酸镁对人脐静脉血管内皮细胞照射后NF-κB/ICAM-1的影响

辐射防护 ›› 2019, Vol. 39 ›› Issue (6): 522-527.

硫酸镁对人脐静脉血管内皮细胞照射后NF-κB/ICAM-1的影响

封士成¹, 徐华林², 孙阳³, 徐瑞君⁴, 崔凤梅⁴, 涂彧⁴

1.东南大学附属中大医院,南京 210000;
2.徐州医科大学附属淮安医院, 江苏淮安 223002;
3.张家港市第一人民医院,江苏张家港 215600;
4.苏州大学放射医学与防护学院,江苏苏州 215213

收稿日期:2019-03-31 出版日期:2019-11-20 发布日期:2020-05-15
通讯作者: 涂彧。E-mail:tuyu@suda.edu.com
作者简介:封士成(1984—),男,2014年毕业于苏州大学放射医学专业,住院医师。E-mail: 2465041441@qq.com

The influence of MgSO₄ on the NF-κB/ICAM-1 of human umbilical vein endothelial cells after irradiation

FENG Shicheng¹, XU Hualin², SUN Yang³, XU Ruijun⁴, CUI Fengmei⁴, TU Yu⁴

1. Zhongda Hospital Affiliated to Southeast University, Nanjing 210000;
2. The Affiliated Huai’an Hospital of Xuzhou Medical University, Jiangsu Huaian 223002;
3. Zhangjiagang First People’s Hospital, Jiangsu Zhangjiagang 215600;
4. School of Radiation Medicine and Protection, Medical School of Soochow University, Jiangsu Suzhou 215213

Received:2019-03-31 Online:2019-11-20 Published:2020-05-15

摘要/Abstract

摘要： 为研究硫酸镁对粘附蛋白ICAM-1及其调控基因NF-κB表达的影响,进而探讨硫酸镁对人脐静脉血管内皮细胞(human umbilical vein endothelial,HUVEC)放射损伤的防护机制。取对数生长期细胞分为空白对照组、单纯照射组、照射加药组,单次4 Gy X射线照射单纯照射组及照射加药组,其中照射加药组于照射前0.5 h加入1.25 mg／mL硫酸镁,收集样本,利用流式细胞术和RT-PCR法检测NF-κB、ICAM-1 mRNA的表达情况,Western blot、激光共聚焦检测NF-κB、ICAM-1蛋白的表达情况。结果表明,照射后72 h内,NF-κB、ICAM-1 mRNA表达均高于空白对照组,其中照后3 h表达最为显著;在照射后48、72 h,1.25 mg/mL硫酸镁处理组中NF-κB / ICAM-1蛋白均低于单纯照射组。以上结果说明,电离辐射能够诱导NF-κB /ICAM-1高表达,而硫酸镁能抑制其表达,且可能是通过抑制NF-κB核转位来调控ICAM-1表达的途径参与辐射损伤防护。

关键词: 硫酸镁, 人脐静脉血管内皮细胞, 照射, ICAM-1, NF-κB

Abstract: To study the effect of magnesium sulfate on the expression of NF - κ B and ICAM-1, and to explore the protective mechanism of magnesium sulfate on radiation injury of HUVECs. The cells under exponential growth phase were divided into control group, irradiation group and irradiation+MgSO₄ group, and treated with a single 4 Gy X-ray except the control group, while 1.25 mg/mL was added to the irradiation+MgSO₄ group 0.5 h before the irradiation. Flow cytometry analysis and RT-PCR were used to study the expression of NF-κB/ICAM-1 mRNA, and NF-κB/ICAM-1 protein expression was detected by Western blot and the laser confocal microscope. The results showed that within 72 hours after irradiation, the expression levels of NF-κB、ICAM-1 mRNA were higher than that of control group, especially for 3 h;48 h and 72 h post-irradiation. The protein level of NF-κB/ ICAM-1 Proteins expression in the group of treated by 1.25 mg/mL MgSO₄ was significantly lower than that of the controls. These results indicate that ionizing radiation can induce the over expression of NF-κB/ICAM-1, and MgSO₄ down-regulation the expression of ICAM-1 by inhibited the trans-location of NF-κB, which may be the mechanisms of radiation protection.

Key words: MgSO₄, human umbilical vein endothelial, radiation, ICAM-1, NF-κB

中图分类号:

R142+.4

封士成, 徐华林, 孙阳, 徐瑞君, 崔凤梅, 涂彧. 硫酸镁对人脐静脉血管内皮细胞照射后NF-κB/ICAM-1的影响[J]. 辐射防护, 2019, 39(6): 522-527.

FENG Shicheng, XU Hualin, SUN Yang, XU Ruijun, CUI Fengmei, TU Yu. The influence of MgSO₄ on the NF-κB/ICAM-1 of human umbilical vein endothelial cells after irradiation[J]. RADIATION PROTECTION, 2019, 39(6): 522-527.

参考文献 20

[1]	贾庆明, 罗海清, 余忠华. 放射性脑损伤发病机制与治疗方法研究进展[J]. 中华实用诊断与治疗杂志, 2018, 32(12):1236-1239.JIA Q M, LUO H Q, YU Z H. Pathogenesis and treatment of radiation-induced brain injury[J]. Journal of Chinese Practical Diagnosis and Therapy, 2018, 32(12):1236-1239.
[2]	MA Z C, HONG Q, WANG Y G, et al. Ferulic acid attenuates adhesion molecule expression in gamma-radiated human umbilical vascular endothelial cells[J]. Biological & Pharmaceutical Bulletin, 2010, 33(5):752-758.
[3]	CHEN N, XU R J, WANG L L, et al. Protective effects of magnesium sulfate on radiation induced brain injury in rats[J]. Curr Drug Deliv, 2018,15(8):1159-1166.
[4]	徐华林. 硫酸镁防治人脐静脉血管内皮细胞放射损伤机制的初步探索[D]. 苏州大学, 2015.XU H L. The exploration of the mechanism MgSO4 prevent and treat the radiation-induced HUVEC injury[D]. Soochow University, 2015.
[5]	Pena L A, Fuks Z, Kolesnick R N. Radiation-induced apoptosis of endothelial cells in the murine central nervous system: protection by fibroblast growth factor and sphingomyelinase deficiency[J]. Cancer Research, 2000, 60(2):321-327.
[6]	LI Y Q, CHEN P, Jain V, et al. Early radiation-induced endothelial cell loss and blood-spinal cord barrier breakdown in the rat spinal cord[J]. Radiation Research, 2004, 161(2):143-152.
[7]	Rousseau M, Marie-Hélène Gaugler, Rodallec A, et al. RhoA GTPase regulates radiation-induced alterations in endothelial cell adhesion and migration[J]. Biochemical & Biophysical Research Communications, 2011, 414(4):750-755.
[8]	Benjaminsen I C, Graff B A, Brurberg K G, et al. Assessment of tumor blood perfusion by high-resolution dynamic contrast-enhanced MRI: A preclinical study of human melanoma xenografts[J]. Magnetic Resonance in Medicine, 2004, 52(2):269-276.
[9]	Tanino T, Kanasaki Y, Tahara T, et al. Radiation-induced microbleeds after cranial irradiation: evaluation by phase-sensitive magnetic resonance imaging with 3.0 tesla[J]. Yonago Acta Medica, 2013, 56(1):7-12.
[10]	MING W, HONG L, HUANG H, et al. Increased expression of EMMPRIN and VEGF in the rat brain after gamma irradiation[J]. Journal of Korean Medical Science, 2012, 27(3):291-299.
[11]	Wilson C M, Gaber M W, Sabek O M, et al. Radiation-induced astrogliosis and blood-brain barrier damage can be abrogated using anti-TNF treatment[J]. International Journal of Radiation Oncology Biology Physics, 2009, 74(3):934-941.
[12]	Brown W R, Blair R M, Moody D M, et al. Capillary loss precedes the cognitive impairment induced by fractionated whole-brain irradiation: A potential rat model of vascular dementia[J]. Journal of the Neurological Sciences, 2007, 257(1-2):67-71.
[13]	Benekli M, Güllü I H, Tekuzman G, et al. Circulating intercellular adhesion molecule-1 and E-selectin levels in gastric cancer[J]. Br J Cancer, 1998, 78(2):267-271.
[14]	Kesanakurti D, Chetty C, Rajasekhar Maddirela D, et al. Essential role of cooperative NF-κB and Stat3 recruitment to ICAM-1 intronic consensus elements in the regulation of radiation-induced invasion and migration in glioma[J]. Oncogene, 2013, 32(43):5144-5155.
[15]	Behrends U, Peter R U, Hintermeier-Knabe R, et al. Ionizing radiation induces human intercellular adhesion molecule-1 in vitro[J]. Journal of Investigative Dermatology, 1994, 103(5):726-730.
[16]	Kim S, Joo Y E. Theaflavin inhibits LPS-induced IL-6, MCP-1, and ICAM-1 expression in bone marrow-derived macrophages through the blockade of NF-κB and MAPK signaling pathways[J]. Chonnam Medical Journal, 2011, 47(2): 104-110.
[17]	LIU X H, PAN L L, YANG H B, et al. Leonurine attenuates lipopolysaccharide-induced inflammatory responses in human endothelial cells: Involvement of reactive oxygen species and NF-κB pathways[J]. European Journal of Pharmacology, 2012, 680(1-3):108-114.
[18]	Baeuml H, Behrends U, Peter R U, et al. Ionizing radiation induces, via generation of reactive oxygen intermediates, intercellular adhesion molecule-1 (ICAM-1) gene transcription and NFκB-like binding activity in the ICAM-1 transcriptional regulatory region[J]. Free Radical Research Communications, 1997, 27(2): 127-142.
[19]	Hoane M R. Assessment of cognitive function following magnesium therapy in the traumatically injured brain[J]. Magnesium Research, 2007, 20(4): 229-236.
[20]	Rochelson B, Dowling O, Schwartz N, et al. Magnesium sulfate suppresses inflammatory responses by human umbilical vein endothelial cells (HUVECs) through the NFkappaB pathway[J]. Journal of Reproductive Immunology, 2007, 73(2):101-107.