RESUMEN
Bone marrow mesenchymal stem cells (MSCs) are considered as a promising cell source to treat the acute myocardial infarction. However, over 90% of the stem cells usually die in the first three days of transplantation. Survival potential, migration ability and paracrine capacity have been considered as the most important three factors for cell transplantation in the ischemic cardiac treatment. We hypothesized that stromal-derived factor-1 (SDF-1)/CXCR4 axis plays a critical role in the regulation of these processes. In this study, apoptosis was induced by exposure of MSCs to H(2)O(2) for 2 h. After re-oxygenation, the SDF-1 pretreated MSCs demonstrated a significant increase in survival and proliferation. SDF-1 pretreatment also enhanced the migration and increased the secretion of pro-survival and angiogenic cytokines including basic fibroblast growth factor and vascular endothelial growth factor. Western blot and RT-PCR demonstrated that SDF-1 pretreatment significantly activated the pro-survival Akt and Erk signaling pathways and up-regulated Bcl-2/Bax ratio. These protective effects were partially inhibited by AMD3100, an antagonist of CXCR4.We conclude that the SDF-1/CXCR4 axis is critical for MSC survival, migration and cytokine secretion.
Asunto(s)
Animales , Masculino , Ratas , Apoptosis , Proteínas Reguladoras de la Apoptosis , Genética , Metabolismo , Células de la Médula Ósea , Metabolismo , Fisiología , Hipoxia de la Célula , Movimiento Celular , Quimiocina CXCL12 , Genética , Farmacología , Fisiología , Citocinas , Metabolismo , Expresión Génica , L-Lactato Deshidrogenasa , Metabolismo , Sistema de Señalización de MAP Quinasas , Células Madre Mesenquimatosas , Metabolismo , Fisiología , Proteínas Proto-Oncogénicas c-akt , Metabolismo , Ratas Sprague-Dawley , Receptores CXCR4 , MetabolismoRESUMEN
This study was amied to construct CXCR4 gene modified bone marrow mesenchymal stem cells (MSCs), and investigate the effect of CXCR4 expression on MSCs migration. The retrovirus vector pMSCV-CXCR4-IRES-GFP that expresses human CXCR4 gene was cloned,the MSCs were transduced by the virus, and the expression of OXCR4 was analyzed by FACS, RT-PCR and immunofluorescence staining. The migration assay was performed using Transwell method in the presence of SDF-1. FACS results showed that 46% of the transduced MSCs were CXCR4 positive, and 57% were GFP positive. The expression of CXCR4 in MSCs was also confirmed by RT-PCR and immunostaining. The migration of MSCs was induced by SDF-1 and was strongly dependent on CXCR4 expression. The concentration of SDF-1 had effect on the migration and the transmigration rate of CXCR4 modified; the amount of MSCs was 5-fold higher than that of untransduced MSCs when the optimal concentration rose to 50 ng/ml. These data indicate that SDF-1/CXCR4 plays an important role in MSCs migration ,and the CXCR4 genetic modification approach could be applied to enhance cell homing, and engraftment in MSCs therapy.
Asunto(s)
Humanos , Células de la Médula Ósea , Biología Celular , Movimiento Celular , Células Cultivadas , Quimiocina CXCL12 , Farmacología , Vectores Genéticos , Genética , Células Madre Mesenquimatosas , Biología Celular , Metabolismo , Receptores CXCR4 , Genética , Retroviridae , Genética , Metabolismo , Transducción Genética , TransfecciónRESUMEN
Objective To investigate the adsorption function of macroporous adsorption resins for the separation and purification of the effective components in Plumula Nelumbinis. Methods AKS-W, AB-8, and H214 resins were systematically studied for their adsorption capability, and the contents of the liensinine, isoliensinine, and neferine were determined by HPLC. Results AKS-W Resin had a preferable adsorptive and separateive effect for the effective components in Plumula Nelumbinis. The products containing 10.2% liensinine, 6.7% isoliensinine, and 11.9% neferine could be obtained. Conclusion Macro-porous-type and non-polar adsorption resin AKS-W is an ideal adsorbent for the extraction of the effective components in Plumula Nelumbinis.