| Objective: In this experiment, recombinant adeno-associated virus mediated into basic fibroblast growth factor(bFGF), mesenchymal stem cells(MSCs), gene modified mesenchymal stem cells were introduced to the skeletal muscle blunt trauma rats seperately.Then we explored the change of bFGF,Collagen-Ⅰ,Collagen-ⅢmRNA and erum creatine kinase expression level histologically and biochemically. Finally, we compared the effects of different treatments and enriched the research data.Methods:100 male ICR rats were randomly divided into 5 groups. Group A was set to the control group. For the rest of the group, set the skeletal muscle in rats after acute contusion : the natural healing group(group B), cell group(group C), genome group(group D), transgenetic cell group(group E).Each group were set up for 3 days,7 days, 14 days and 21 days(5 rats in each group) respectively. Except for the control group, the rest of the group received local injections immediately,such as PBS(phosphate buffer saline), MSCs, rAAV2- bFGF and transfection rAAV2- bFGF MSCs after the injury in the muscle damage. Tracking test groups at different time points. Following indicators were skeletal muscle tissue morphology, tissue damage bFGF expression, collagen I, IIImRNA, and the changes of serum creatine kinase.Results: 1) HE staining light microscope and the changes of plasma CK activity indicated that mesenchymal stem cells and basic fibroblast growth can promote muscle regeneration after the rats with acute skeletal muscle contusion, reduce scar formation,accelerated injury muscle healing processes, improved muscle healing.2) The expression of bFGF was significantly higher than that in group A(P<0.05)after injury. With the repairing time increased, the bFGF of each group showed a downward trend. The expression of bFGF in group E was significantly higher than theexpression level in group A at the 21 st day. The amounts of bFGF from high to low were demomstrated as group E, group C, group D, group B and group A. 3) The expression of collagen I mRNA was increased in each group after injury. After skeletal muscle injury, the expression of collagen I mRNA increased slightly. After seven days the injury continues to rise, at the fourteenth day, the expression of collagen I mRNA increased rapidly, and reached the peak in the 21 st day. The expression of collagen ImRNA was the lowest in group E. Compared with the normal control group, except for the third day, significant differences were found among each group(P<0.01).4) The expression of collagen ⅢmRNA was increased and then decreased, and reached the peak in 7th day. The expression of collagen protein ⅢmRNA from high to low was group B, group D, group C, group E and group A. The phenomemon above demonstrates that fibroblast growth factor(bFGF) and mesenchymal stem cells(MSCs)in skeletal muscle accelerated the healing of the injury by meaning of inhibiting the expression of Collagen-Ⅰ,Collagen-ⅢmRNA in rats’ tissue.Conclusions : 1) Successfully packaged rAAV r AAV2-bFGF, after transfection of mesenchymal stem cells express a reporter gene, the gene of bFGF also highly expressed.2) Mesenchymal stem cells and basic fibroblast growth factor are capable of promoting the expressin of bFGF in skeletal muscle, which is able to inhibit the expression of Collagen-Ⅰ,Collagen-ⅢmRNA in local tissues as injured skeletal muscle,and reduce the formation of scar. 3) Over time, the effect of treatment in each group the same general trend.4) Transgenetic mesenchymal stem cells therapy can promote the repair of injured skeletal muscle, and it is more efficient than that of pure stem cell therapy or gene therapy. |
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