RESUMO
BACKGROUND: The cel density is one of the factors involved in the state of cel differentiation, and the effect of cel density on transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells is stil unclear. OBJECTIVE: To observe the effect of cel density on transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells. METHODS: HaCaT cells was seeded in 6-wel plates at low density of 103/cm2 and high density of 105/cm2 then treated by 2 μg/L transforming growth factor-β1 for 48 hours, thereafter observed the changes in cel morphology. The transcription levels of epithelial cadherin, tight junction protein-1, vimentin, neuronal-cadherin were detected by real-time PCR, and expression levels of epithelial cadherin and vimentin were detected by Western blot. RESULTS AND CONCLUSION: Cel gap of HaCaT cells grew larger after treated with transforming growth factor-β1 for 48 hours, and cel morphology was long spindle rather than polygonal in low-density group, while in high-density group without obvious morphological changes. The real-time PCR showed that the transcriptions of epithelial cel marker epithelial cadherin and tight junction protein-1 were suppressed when compared with the control group (P < 0.05), and the decreasing deree in the high-density group was higher than that in the low-density group (P <0.05), mesenchymal cel marker neuronal-cadherin and vimentin were upregulated in the high-density group and the low-density group when compared with the control group (P < 0.05), and there were no significant differences between high-density group and low-density group. Western blot results verified the changes of neuronal-cadherin and vimentin expression level. These results suggest that the high seeding density can inhibit transforming growth factor-β1-induced epithelial-mesenchymal transition of HaCaT cells.
RESUMO
Objective To study the effects of bone marrow mesenchymal stem cells transplantation on functional recovery of the injured rats spinal cord.Methods MSCs labeled with Brdu were transplanted into rats model of spinal cord half-transection injury.The open-field BBB scoring system was employed to evaluate behavioral changes.MSCs' survival after transplantation was identified by BrdU immunohisto chemistry.We observed the reconstruction of neuronal circuits by HRP coloration.The recovery of transduction function after spinal injury was examined by cortex somatosensory evoked potential (CSEP).Results Treated rats generally showed better functional recovery than control rats after operation.BrdU-positive cells could be found in the spinal cord injury site one week after transplantation.At two months after transplantation, HRP-positive cells could be found at rostral of the spinal cord injury site of treated rats, but not be found in control.CSEP could be evoked at treated rats from two months after transplantation,but not in controls.Conclusion MSCs may survive in the spinal cord injury site via local injection immediately after spinal cord injury, and may promote regeneration of the injured axons.