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

[超低周波電磁界で処置されたヒト強膜線維芽細胞におけるI型コラーゲンの抑制] med./bio.

Suppression of type I collagen in human scleral fibroblasts treated with extremely low-frequency electromagnetic fields.

掲載誌: Mol Vis 2013; 19: 885-893

この研究は、超低周波電磁界(ELF-EMF)下で、網膜色素上皮(RPE)細胞培養上清で処理されたヒト胎児強膜線維芽細胞(HFSF)におけるI型コラーゲン(COL1A1)の発現を調べた。処理されたHFSFの増殖は、細胞カウントキット-8アッセイで評価した。処理されたHFSFにおけるCOL1A1およびマトリックスメタロプロテイナーゼ-2(MMP-2)の発現は逆転写PCR(RT-PCR)およびウエスタンブロットで検出した。また、EMFばく露RPE細胞における形質転換成長因子-beta2(TGF-beta2)および塩基線維芽細胞成長因子-2(FGF-2)の発現はRT-PCRで検出した。さらに、HFSFにおけるCOL1A1およびMMP-2の発現は、免疫蛍光染色によっても確認した。HFSFにおける細胞シグナル調節キナーゼ1/2(ERK1/2)およびp38の活性化をウエスタンブロットで測定した。その結果、ELF-EMFばく露によりHFSFの増殖率が低下すること、さらにRPE培養上清の添加により、この効果は増強される可能性が示された;対照細胞と比較して、ELF-EMF下のHFSFではコラーゲン合成の有意な減少が検出された;しかし、MMP-2の発現は増加すること、この増加は、RPE培養上清の添加によりさらに増強されることが示された;ERK1/2およびp38の活性は、ELF-EMFばく露を受けたHFSFで有意に増加し、この増加はRPE培養上清の添加によりさらに増強される可能性が示された、と報告している。

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研究目的(著者による)

To examine the effects of an exposure to extremely low frequency magnetic fields on the collagen synthesis in human fetal scleral fibroblasts.

詳細情報

The sclera is a part of the human eye ("white of the eye"), mainly composed of collagen fibrils. Several factors can influence the correct formation and structure of these collagen fibrils. Therefore, the expression of several genes related to the collagen synthesis was examined.
The retina is the source of ocular growth-regulating signals and retinal pigment epithelial cells are intimately involved in eye growth regulation. For this reason, supernatant from separately pre-exposed retinal pigment epithelial cells was added to the scleral fibroblasts during the exposure.

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: continuous for 24 hours

ばく露1

主たる特性
周波数 50 Hz
タイプ
  • magnetic field
波形
  • sinusoidal
ばく露時間 continuous for 24 hours
ばく露装置
ばく露の発生源/構造
ばく露装置の詳細 magnetic fields were generated in a device constructed by winding 150 turns of insulated soft copper wire with a diameter of 1.5 mm; three horizontally parallel Helmholtz coils (16 cm length, 14 cm width, 8 cm height) and one slidac; coils were placed vertically, facing one another; exposure system was put in a humidified incubator at 37 °C, which contained 5% CO2; cells were placed between the coils
Sham exposure A sham exposure was conducted.
パラメータ
測定量 種別 Method Mass 備考
磁束密度 0.2 mT - 測定値 - -

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:
調査の時期:
  • ばく露後

研究の主なアウトカム(著者による)

Magnetic field-exposed cell cultures of scleral fibroblasts showed a significant reduced proliferation rate compared to not-exposed ones. An addition of the supernatant of retinal pigment epithelial cells led again to a significant decrease in the proliferation rate compared to those exposed to the magnetic field only.
In exposed scleral fibroblasts, the expression of collagen I was significantly decreased compared to the control cell cultures, while the expression of matrix-metalloproteinase-2 was significantly increased. This effect could be significantly enhanced through the addition of retinal pigment epithelial cell supernatant. In comparison to the control cell culture of scleral fibroblasts, the levels of ERK1/2 and p38 MAP kinase were significantly increased in the exposed cell cultures, while an addition of retinal pigment epithelial cell supernatant led again to a significant increase compared to those exposed to magnetic fields alone.
The authors conclude that exposure to extremely low frequency magnetic fields can influence the collagen synthesis in human scleral fibroblasts and hence could disturb the formation of the sclera.

研究の種別:

研究助成

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