To study the effects and the mechanism of a pulsed electric field on insulin and its subsequent mediation of proliferative changes in human hepatocytes in vitro.
Insulin solution was exposed to pulsed electromagnetic field and added to the culture medium of hepatocytes. Insulin is usually added into the culture medium to accelerate cell growth.
| Exposure | Parameters |
|---|---|
|
Exposure 1:
50 kHz
Modulation type:
pulsed
Exposure duration:
continuous for 10 min, 20 min and 40 min
|
|
| Frequency | 50 kHz |
|---|---|
| Type | |
| Waveform | |
| Exposure duration | continuous for 10 min, 20 min and 40 min |
| Modulation type | pulsed |
|---|---|
| Pulse width | 20 µs |
| Rise time | 12 µs |
| Repetition frequency | 50 Hz |
| Exposure source |
|
|---|---|
| Setup | 96-well micro-plate containing insulin stock solution was placed between the two plates, which were 16 mm apart. |
| Sham exposure | A sham exposure was conducted. |
| Measurand | Value | Type | Method | Mass | Remarks |
|---|---|---|---|---|---|
| electric field strength | 7.1 V/m | maximum | estimated | - | - |
Pulsed electric fields produced a conformational change of insulin molecule. The binding capacity of insulin to its receptors was reduced to 87% of the control level after pulsed electric field exposure. The average intracellular tyrosine phosphorylation level decreased by 11%. The expression of 55 of 12000 genes investigated was modified, including an increase in the expression of human tyrosine phosphatase and the small GTP-binding protein.
Based on these data, a mechanism is proposed to explain the biological effects of pulsed electric fields on hepatocyte proliferation through the insulin signaling pathway.
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