Influence Of Acrylamide On NFκB Pathway In BMMSCs From Porcine

It was reported that BMMSCs have the capability of immunoregulation, and participate in process of multiple immune response. It was attracted the attention of researchers that these cells have priority to move to the damaged tissue, so as a kind of potential therapeutic tool for tissue repair. At present, the formation of BMMSCs-isolated method are base on Friedenstein'researches. There is no clear, uniform evaluation standard in the idantification of BMMSCs, it can but depends on the comprehensive analysis of several aspects. Some reports demonstrated ACR was as a probable human carcinogen, neurotoxic, and mutagenic. In present study, we established a BMMSCs culture system, and studied on the influence of ACR disposed on BMMSCs. It hopes that the conclusions could provide a certain clues to research the influence mechanism of ACR.In this experiment, BMMSCs were isolated from 0-6 month-old male Wuzhishan miniature pig, and its replicative capacity waws detected using CCK-8. The identification of BMMSCs in several parts:cell morphology, detections of several aurface antigens of CD34, CD45, CD90, CD29 and CD44. After BMMSCs disposed with 0.5mM ACR for 72h, the ROS, Hsp27 and NFκB using ROS(?) Assay Kit, ELISA and Dual-Luciferase(?) Reporter Assay System, apoptosis and cell cycle were determined respectively.The resultd showed:the shape of BMMSCs isolated from WZSP was spindle or polygon, look like the shape of mechanocyte. The results of staining with Oil red O and alizarin red S are both positive after 21 days adipose and osteogenesis inducing culture. Flow cytometric analysis result showed that CD90, CD29 and CD44 were expressed on BMMSCs, but surface antigens of hematopoietic stem cells CD34 and leucocyte antigen CD45 are not expressed. After BMMSCs disposed with 0.5mM ACR for 72h, we discovered that the expression of ROS and Hsp27 increased (P< 0.05), and NFκB was activated. However, the cell cycle and apoptosis are not obvious alteration.In this paper, we established a optimum culture system for BMMSCs from WZSP. Cells that isolated from bone marrow are indentified by analysising of cell morphology, differentiation, several aurface antigens. When BMMSCs stimulated with low concentrations of ACR (0.5mM), the expression of ROS, Hsp27 and IL-8 increased, and NFκB activated, while the apoptosis and cell cycle have no obvious alteration.