ABSTRACT
[Objective] To observe the effect of extract of Carapax et Plastrum Testudinis (ECPT) on nuclear receptor in the proliferation of rat mesenchymal stem cell (MSC) in vitro. [Methods] The rat MSC dissociated from bone marrow by density gradient method were cultured and identified by marking of bromodeoxyuridine (Brdu) and staining of CD44. Then different doses of ECPT (3333, 333.3 and 33.33 ?g/mL) were respectively added into in-vitro cultured MSC for 12, 24, 72 and 120 hours. The expression of retinoic acid receptor-?(RAR?), vitamin D receptor (VDR), estrogen receptor (ER), glucocorticoid receptor (GR), thyroid hormone receptor-?(TR?) and peroxisome proliferator-activated receptor ?(PPAR?) in MSC was detected by immunohistochemistry and immunofluorescence methods. [Results] The number of RAR?-and VDR-positive cells in ECPT groups was higher than that in the control group (P
ABSTRACT
[Objective] To observe the in-vitro inductive effect of ginsenosides (GS) on differentiation of rat bone marrow mesenchymal stem cells (MSC) into myocardial cells. [Methods] Marrow stromal cells were isolated from adult SD rats, and then were cultured and cloned by density gradient method and adhesive cultivation to prepare the primary MSC. The cell surface antigens of CD34 and CD44 were detected with flow cytometer to identify MSC. After subculture, the 8th generation of continuous MSC were induced by GS (250 mg/L) and 5-aza (5-azacytidine, 10 ?mol/L) and GS + 5-aza (250mg/L and 10 ?mol/L respectively) for 24, 48 and 72 hours to differentiate to myogenic cells. Meanwhile, a blank control group was set to compare the effect. After the induction, myocardial cell percentage was worked out after examining MSC count and positive cells count under the phase contrast microscope. The specific proteins of Desmin and cardiac troponin I (cTnI) in myocardial cells were detected by immunohistochemistry method, and the cardiac-specific gene expression of ?-MHC (myosin heavy chain) and ?-MHC in myocardiocytes was examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis. [Results] The cultured marrow stromal cells grew well, spindle in shape, and their CD44 expression was positive, indicating that marrow stromal cells differentiated to MSC. After induction with GS and 5-aza, MSC differentiated to myocardial cells, and the myocardial cell percentage was 38% in GS group, 35% in 5-aza group and 39% in GS + 5-aza group, the difference being insignificant. The expression of Desmin, cTnI and MHC in MSC was positive, and MSC were spindle-shaped and looked like fibroblast, indicating that MSC differentiated to myocardial cells. [Conclusion] Ginsenosides can induce MSC to differentiate to myogenic cells in-vitro, and this will supply evidence for cell transplantation.