研究のタイプ: 医学/生物学の研究 (experimental study, observational study)

[血管内皮の抗凝固性および凝固促進性に対する低周波電界の影響:高電圧電撃傷への新しい洞察] med./bio.

Influence of low frequency electric fields on anti- and pro-coagulability of the vascular endothelium: new insights into high-voltage electrical injury.

掲載誌: Thromb Haemost 2004; 91 (5): 1000-1008

この研究は、高電圧電撃傷から96時間後の患者(n = 3)において、インビボで、TAT、F1+2、PAI-1、およびt-PAの血漿レベルを測定した。またインビトロでは、50 Hz、60 V/ cm、持続時間20 msの電界に1、10、30、および50周期のばく露を受けたHUVECsの培養細胞の上清で、ばく露後の72時間、タンパク質S、TF、ET-1、PGI2NOt-PA、およびPAI-1を測定した。これは、内皮細胞損傷が関連する止血への影響を分析するためである。さらに、トロンボモジュリン発現免疫組織化学的に分析した。その結果、F1+2とTATのレベル上昇で示される凝固の活性化が損傷後12〜72時間の患者で観察されたが、その一方、PAI-1が高いために線維素溶解は妨げられた;1人の患者は、3日目までに血管の血栓症を呈した;インビトロでは、ばく露から2〜48時間において、30〜50周期のばく露を受けたHUVECsの培地で、PAI-1が有意に上昇した(p <0.05);4〜72時間においては、10、30、および50回ばく露を受けたHUVECsの培地で、t-PAの濃度が有意に低下した(p <0.05)が、その一方、30および50周期のばく露を受けた細胞群のTFの濃度が上昇した;ばく露から24、48、および72時間後、30および50周期のばく露を受けたHUVECsでトロンボモジュリンの染色は弱いか無しであった;インビトロ実験でばく露後に見られる凝固系と線維素溶解のバランスの乱れは、組織損失につながる電撃傷後の血管の進行性血栓症の臨床的観察を説明するかもしれない、と報告している。

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研究目的(著者による)

After high-voltage electric injury, patients often show tissue necrosis and thrombosis of blood vessels even remote from entry and exit site of electrical current.
In this study, blood coagulation markers and fibrinolytic avtivity (plasma levels of thrombin-antithrombin (TAT), prothrombin fragment 1+2 (F1+2), tissue-type plasminogen activator (t-PA) and its inhibitor PAI-1 (type-1 plasminogen activator inhibitor)) were determined in vivo in three patients with high-voltage injury for 96 hours after trauma.
In order to examine a possible effect on haemostasis related to endothelial cell damage, protein S, tissue factor (TF), endothelin-1 (ET-1), prostacyclin (PGI2), nitric oxide (NO), t-PA, and PAI-1 were determined for 72 hours in vitro in cell culture supernatant of human umbilical vein endothelium cells (HUVECs) that had been exposed to 1, 10, 30, and 50 electric field periods of 50 Hz with field strength of 60 V/cm and duration of 20 ms. Furthermore, expression of thrombomodulin was determined.

影響評価項目

ばく露

ばく露 パラメータ
ばく露1: 50 Hz
ばく露時間: 20 ms periods interupted by 10 s intervals, 1, 10, 30 and 50 periods

ばく露1

主たる特性
周波数 50 Hz
タイプ
  • electric current
波形
  • sinusoidal
ばく露時間 20 ms periods interupted by 10 s intervals, 1, 10, 30 and 50 periods
ばく露装置
ばく露の発生源/構造
  • electrode
チャンバの詳細 culture chamber (9 x 9 x 2 cm lxwxh) with 4 mm of culture medium maintained at 37°C
ばく露装置の詳細 Electrodes were placed on the inside of the cover of the culture plate at two opposite sides
Sham exposure A sham exposure was conducted.
パラメータ
測定量 種別 Method Mass 備考
電界強度 60 V/cm unspecified 指定なし - -

ばく露を受けた生物:

方法 影響評価項目/測定パラメータ/方法

研究対象とした生物試料:
研究対象とした臓器系:
  • vascular system
調査の時期:
  • ばく露後

研究の主なアウトカム(著者による)

Blood clotting activation could be found in the patients by increased levels of F1+2 and TAT between 12 and 72 hours after injury, whereas fibrinolysis was disturbed due to high PAI-1. One patient presented thrombosis of vessels by day 3.
In vitro, PAI-1 increased significantly in medium of endothelium cells with an application of 30 and 50 periods between 2 and 48 hours after exposure. Between 4 and 72 hours, the concentration of t-PA was significantly lower in the medium of endothelium cells exposed to 10, 30, and 50 periods, whereas there was a significant increase in the concentration of TF in the cell groups with an application of 30 and 50 periods. 24, 48, and 72 hours after injury (exposure), there was just weak or no staining for thrombomodulin in the cells with an application of 30 and 50 periods.
The disturbed balance between blood clotting system and fibrinolysis observed in vitro after electric injury might explain the clinical observation of a progressive thrombosis of blood vessels after electric injury leading to tissue loss.

研究の種別:

研究助成