この研究は、ヒト羊膜上皮細胞のミトコンドリアの膜透過性に対する50Hz磁界ばく露（MF：0.4mT、ばく露時間は変化させた）の影響を調べた。その結果、60分間ばく露により、ミトコンドリア膜透過性遷移（MPT）とシトクロムcの放出が一過性に誘導されたが、ミトコンドリア膜電位への有意な影響はなかった；総Bcl-2結合Xタンパク質レベルを除き、ミトコンドリアのBaxおよびBcl-2のレベルへの影響は見られなかった；MFばく露を受けた細胞では、細胞内活性酸素種（ROS）レベルおよびグリコーゲン合成キナーゼ-3β（GSK-3β）のセリン残基での脱リン酸化が増加した；ROSスカベンジャまたはGSK-3β阻害剤により、MF誘導性MPTは減少した、と報告している。
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The effects of exposure to a 50 Hz magnetic field on mitochondrial permeability and reactive oxygen species should be investigated in human amniotic epithelial cells.
The mitochondrial permeability transition (MPT), which is defined as an increase in the permeability of the mitochondrial membranes for small molecules via opening of so called MPT pores, was examined among other parameters.
Cells were either exposed or sham exposed and some samples were also treated with acetylcysteine (a ROS scavenger), SB216763 (an inhibitor of glycogen synthase kinase (GSK)-3-beta, which might play a role in MPT), and with cyclosporine A (a MPT pore inhibitor).
| 周波数 | 50 Hz |
|---|---|
| タイプ |
|
| 波形 |
|
| ばく露時間 | continuous for 5, 15, 30, 60 or 120 minutes |
| ばく露の発生源／構造 | |
|---|---|
| チャンバの詳細 | two exposure chambers in incubator with 95% air humidity and 5% CO2; the temperature was kept at 37.0 ± 0.2°C |
| ばく露装置の詳細 | each chamber contained a set of square Helmholtz coils (20 cm Œ 20 cm), which were double-wrapped with two lines of copper wire and encased by µ-metal to shield cells from stray fields; a fan in the wall made air and temperature uniform between chambers and incubator; cell dishes were put in the center of the coils; the magnetic field was perpendicular to the dishes and heterogeneity of MF distribution within the 12 x 12 x 20 cm3 space in the exposure chamber was less than 1% |
| Sham exposure | A sham exposure was conducted. |
| Additional information | one chamber was used for sham exposure with opposite direction currents fed into the coils; exposure and the corresponding sham exposure was conducted simultaneously; the difference of temperature between exposure and sham exposure did not exceed 0.1°C |
| 測定量 | 値 | 種別 | Method | Mass | 備考 |
|---|---|---|---|---|---|
| 磁束密度 | 4 mT | - | 測定値 | - | - |
Exposure to the magnetic field for 60 min led to a significant increase of MPT compared to sham exposed cells while a simultaneous treatment with cyclosporine A prevented this effect. The mitochondrial membrane potential was not affected by exposure to the magnetic field.
The cytosolic protein expression of cytochrome-c was significantly increased and the whole cell protein expression of Bax was significantly decreased after exposure for 60 minutes compared to sham exposed cells. Protein expression of phosphorylated GSK-3-beta was significantly increased after 15 minutes and significantly decreased after 60 minutes of exposure compared to sham exposed cells. A treatment with the GSK-3-beta inhibitor SB216763 prevented MPT after 60 minutes of exposure. The level of reactive oxygen species was significantly increased after 5, 15 and 30 minutes of exposure compared to sham-exposed cells, with acetylcystein preventing this increase and preventing MPT. Moreover, acetylcystein prevented the changes in protein expression of phosphorylated GSK-3-beta, indicating that magnetic field induced MPT depends on GSK-3-beta activation via reactive oxygen species.
The authors conclude that exposure to a 50 Hz magnetic field might induce mitochondrial permeability transition in human amniotic epithelial cells through the ROS/GSK-3-beta signaling pathway.
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