Medical science monitor basic research · 2013
Effects of verapamil on the immediate-early gene expression of bone marrow mesenchymal stem cells stimulated by mechanical strain in vitro.
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Li Runguang, Wei Mingfa, Shao Jingfan
Аннотация
BACKGROUND: To study the effects of verapamil on the immediate-early genes (IEGs) expression of bone marrow mesenchymal stem cells (MSCs) stimulated by cyclic mechanical strain, in order to deduce the role of calcium ion channel in the cell signaling responses of MSCs to mechanical strain.
MATERIAL AND METHODS: MSCs were isolated and cultured, and the passage of 3-6 MSCs were stimulated by mechanical strain and pretreated with or without verapamil. After that, flow cytometry was used to measure the fluorescence intensity of intracellular Ca(2+) immediately. The expression of early-response genes/proteins (c-fos, c-jun and c-myc) were examined by RT-PCR, immunohistochemistry and Western blot.
RESULTS: Intracellular Ca(2+) concentration of MSCs significantly changed when stimulated by cyclic strain, and the expression of c-fos, c-jun and c-myc remarkably increased in both mRNA and protein levels, while verapamil pre-treatment partially inhibited these effects (P<0.01).
CONCLUSIONS: The changes of the intracellular calcium concentration of MSCs induced by mechanical strain, dependent on the regulation of calcium channel activation, might play a role in the early response of MSCs to cyclic strain.
MATERIAL AND METHODS: MSCs were isolated and cultured, and the passage of 3-6 MSCs were stimulated by mechanical strain and pretreated with or without verapamil. After that, flow cytometry was used to measure the fluorescence intensity of intracellular Ca(2+) immediately. The expression of early-response genes/proteins (c-fos, c-jun and c-myc) were examined by RT-PCR, immunohistochemistry and Western blot.
RESULTS: Intracellular Ca(2+) concentration of MSCs significantly changed when stimulated by cyclic strain, and the expression of c-fos, c-jun and c-myc remarkably increased in both mRNA and protein levels, while verapamil pre-treatment partially inhibited these effects (P<0.01).
CONCLUSIONS: The changes of the intracellular calcium concentration of MSCs induced by mechanical strain, dependent on the regulation of calcium channel activation, might play a role in the early response of MSCs to cyclic strain.
Предметные рубрики MeSH
CalciumCalcium Channel BlockersCells, CulturedChildChild, PreschoolFlow CytometryGene Expression RegulationGenes, Immediate-EarlyHumansIon ChannelsIonsMesenchymal Stem CellsProto-Oncogene Proteins c-fosProto-Oncogene Proteins c-junProto-Oncogene Proteins c-mycSignal TransductionStress, MechanicalVerapamil