Proteomics Analysis Of Effect Of High Frequency Pulsed Electromagnetic Fields On The Proliferation Of Bone Marrow Mesenchymal Stem Cells

Bone marrow mesenchymal stem cells (MSCs) with multilineage differentiation potential、immune tolerance、drawing a convenient、self-renewal characteristics become the important seed cells in tissue engineering and a hot research in recent years. A preliminary study of this subject shows that the high-frequency pulsed electromagnetic fields promote apoptosis of marrow mesenchymal stem cells, but its mechanism is unclear.Proteomics is the process to study and identify the total protein expressed by a cell, organ, organization at a particular time.Proteomics research is arisen, and this has laid a foundation for us to study the mechanism of high-frequency pulsed electromagnetic fields promote myeloid mesenchymal stem cell apoptosis from proteomics.OBJECTIVE:1. To create the electrophoresis maps of fully protein extracted from high-frequency pulse electromagnetic field interfered and normal cultured mesenchymal stem cell in vitro by using two-dimensional electrophoresis (2-DE).2. To find the differentially expressed proteins by using image analysis software to analyze electrophoresis maps;To identify the functional proteins associated with that high-frequency pulsed electromagnetic fields which promote mesenchymal stem cell apoptosis, and to explore the mechanism of action of the functional protein.METHODS:1. bone marrow mesenchymal stem cells isolated、cultured、 passaged and purified by whole bone marrow adherent In vitro;To take the third generation as the target cells.2. With a1×106/cm2density inoculated into a culture bottle bottom covers an area of100cm2,and then divide into two groups (A, B) to culture,group A for the high-frequency pulsed electromagnetic fields in the intervention group, group B for the control group.3. after fully protein extraction from group A and group B, purify protein samples, measure the protein concentration by Bradford Method, Analysis the electrophoresis maps obtained by two-dimensional electrophoresis, find and cut off differentially expressed proteins to enzymatic hydrolysis, find out the functional proteins from the results of mass spectrometry.RESULTS:Ideal images of fully protein extraction from group A and group B can be obtain,19differentially expressed protein spots were obtain,17proteins up-regulate in group A,meanwhile2proteins down-regulate.11different proteins were identifed by mass spectrometry, and one of them which known as the EF-hand domain-containing protein D2(also known as Swiprosin-1) promotes cell apoptosis. CONCLUSION:Ideal images of fully protein extraction from group A and group B can be obtain by two-dimensional electrophoresis (2-DE); The EF-hand domain-containing protein D2may a functional protein associated with that high-frequency pulsed electromagnetic fields promote mesenchymal stem cell apoptosis.