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CC BY-NC-ND 4.0 · TH Open 2017; 01(01): e3-e10
DOI: 10.1055/s-0037-1603926
Original Article
Georg Thieme Verlag KG Stuttgart · New York

The Comparison of the Protective Effects of α- and β-Antithrombin against Vascular Endothelial Cell Damage Induced by Histone in Vitro

Toshiaki Iba
1   Department of Emergency and Disaster Medicine, Juntendo University Graduate School of Medicine, Tokyo, Japan
,
Tetsuya Sasaki
2   Nihon Pharmaceutical Co. Ltd., Research Laboratory, Narita, Japan
,
Kazutoshi Ohshima
2   Nihon Pharmaceutical Co. Ltd., Research Laboratory, Narita, Japan
,
Koichi Sato
3   Department of Surgery, Juntendo Shizuoka Hospital, Juntendo University Graduate School of Medicine, Tokyo, Japan
,
Isao Nagaoka
4   Department of Host Defense and Biochemical Research, Juntendo University Graduate School of Medicine, Tokyo, Japan
,
Jecko Thachil
5   Department of Haematology, Manchester Royal Infirmary, Manchester, United Kingdom
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Publikationsdatum:
28. Juni 2017 (online)

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Abstract

Antithrombin is a promising option for the treatment of sepsis, and vascular endothelium is an important target for this fatal condition. Here, we aimed to evaluate the protective effects of different glycoforms of antithrombin on histone-induced endothelial cell damage and explore the responsible mechanisms in an experimental model in vitro. Endothelial cells were treated in vitro using histone H4 to induce cellular damage. Various doses of either α- or β-antithrombin were used as treatment interventions, and both cell viability and the levels of lactate dehydrogenase (LDH) in the medium were assessed. Endothelial cell damage was also assessed using microscopic examination and immunofluorescent staining with anti-syndecan-4 and anti-antithrombin antibodies. As a result, both glycoforms of antithrombin significantly improved cell viability when administered at a physiological dose (150 μg/mL). Cellular injury as evaluated using the LDH level was significantly suppressed by β-antithrombin at a supranormal dose (600 μg/mL). Microscopic observation suggested that β-antithrombin suppressed the endothelial cell damage more efficiently than α-antithrombin. β-Antithrombin suppressed the intensity of syndecan-4 staining which became evident after treatment with histone H4, more prominently than α-antithrombin. The distribution of antithrombin was identical to that of syndecan-4. In conclusion, both α- and β-antithrombin can protect vascular endothelial cells from histone H4-induced damage, although the effect was stronger for β-antithrombin. The responsible mechanisms might involve the binding of antithrombin to the glycocalyx on the endothelial surface. These results provide a theoretical basis for the application of antithrombin to the prevention and treatment of sepsis-related endothelial damage.

Keywords

antithrombin - glycoform - vascular endothelial cell - glycocalyx - syndecan-4

Contributions of Authors

The study design was performed by T.I. and J.T. I.N. and K.S. were involved in the study conduct. Data collection was performed by T.S. and K.O. Data analysis was performed by T.I. and K.S., and I.N., T.I., and J.T. were involved in drafting and revising of the manuscript. All authors read and approved the final manuscript.


Supplementary Material

 

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