Magnetic Induction Hyperthermia Effects On Vascular Endothelial Cells

Objective:In this investigation, we proposed to study the effect of magnetic induction hypermia on vascular endothelial cell,including proliferation,apoptosis,cell cycle,migration and gene expression.And explored the possible effect of magnetic induction hypermia on vascular repairing and stent endothelialization after vascular stents implanting. Then we can speculate the feasibility of magnetic inductive hypethermia used for prevention and treatmengt of restenosis after PTCA.Methods:(1)The study of effect of magnetic inductive hyperthermia on cell proliferation and activity of HUVEC:Generally cultured HUVEC and heated them with 316L stainless steel stent in the alternating magnetic flied.Then detected cell proliferation rate and activity after continuing to foster for hours. (2) The study of effects of magnetic inductive hyperthermia on apoptosis and cell cycle changing of HUVEC:Doubly stained cells using Annexin-V and PI after hyperthermia, and detected cell apoptosis rate by flow cytometry.Stained cells after hyperthermia with PI,and detectde cell cycle changing by flow cytometry. (3) The study of effects of magnetic inductive hyperthermia on HUVEC migration:scratched cells,then observed healing of the scratch.Counted the number of migrated cells using Transwell migration method to study cell migration changing. (4)The study of effects of magnetic inductive hyperthermia on HUVEC gene expression:Detected the relative expression level of heat shock protein 70 (HSP70) by ELISA.And used Westernbloting to study HSP70 expression after heating with different temperature.Results:(1)Cell survival rate of cells after 43℃magnetic inductive hyperthermia was 96.12±4.58%,and after 47℃magnetic inductive hyperthermia,the rate is 2.33±2.91%.The former had no significant difference compared with the control group,and there was significant differences between the latter and the control group.(2) The apoptosis rate of cells after 43℃magnetic inductive hyperthermia was 3.93%±1.03%, having no significant difference compared with the control group. The apoptosis rate of cells was significantly increased by 47℃magnetic inductive hyperthermia compared with the control group,having significant differences.And the apoptosis rate was 50.44%±3.12%.43℃magnetic inductive hyperthermia didn't significantly change the cell cycle,but the cell cycle was inhibited by 47℃magnetic inductive hyperthermia,and arrested in GO/G1 phase.(3)After scratches, the number of migrating cells gradually increased in the control group and 43℃magnetic inductive hyperthermia group, and the scratch gradually healed.Cells gradually becamed round and shruken cells increased in the 47℃magnetic inductive hyperthermia group.Also the scratch didn't heal.The number of migrated cells of 43℃magnetic inductive hyperthermia group was 43±2, having no significant difference compared with the control group.But after 47℃magnetic inductive hypermia, the number of migrated cells decreased to 21±4, having significant differences compared with the control group.(4)ELISA test results showed expression of HSP70 in HUVEC was significantly increased after magnetic inductive hyperthermia.And the relative expression was 429.2pg/mL in 43℃magnetic inductive hyperthermia group, as well as it was 536.9pg/mL in 47℃magnetic induction hyperthermia group.Conclusion:(1) 43℃magnetic inductive hyperthermia had no effect on the proliferation and viability of HUVEC,and 47℃magnetic inductive hyperthermia significantly inhibited growth of HUVEC.(2)43℃magnetic inductive hyperthermia could not cause apoptosis and cycle changing of vascular endothelial cells.And 47℃magnetic inductive hyperthermia could increase apoptosis of vascular endothelial cells, also make cell cycle of them arrest in GO/G1 phase.(3)43℃magnetic inductive hyperthermia had no significant effect on migration of vascular endothelial cells,but 47℃magnetic inductive hyperthermia significantly inhibited cell migration. And that suggested 43℃could not hinder vascular repair and stent endothelialization compared with 47℃(4)Expression of HSP70 in HUVEC increased as the heating temperature increased.And that may be one of the mechanisms by which cells protect themselves from being injured by high temperature.