| The effect of hydroxycamptothecin on P-Smad3 expression of hypertrophic scar fibroblasts and hypertrophic scars in rabbit earsPrevention and treatment of pathological scars is a difficult research topic in the Burns and Plastic Surgery. There are many ways to treat pathological scars, as surgery, drugs, radiation, freezing, pressure, etc.The treatments have made some experimental or clinical treatment effects, but the long-term efficacy and satisfaction are far short of the requirements of the patients. Some treatments can even cause serious complications.There bright more greater resistance to the treatment of pathological scars. Therefore, founding a safe and effective treatment of pathological scar is still the difficult tasks which meet the majority of medical science and technology workers. So far, the treatment of pathological scars have not yet made a significant breakthrough. It is closely related to the development of mechanisms of the formation, development, and prognosis of pathological scar.Scar is the over-healing result which is response to the body's injury,including hypertrophic scars and keloids. They are characterized by a large number of fibroblasts proliferation and excessive deposition of extracellular matrix. Currently,the basis research of pathological scars mainly from the areas of fibroblasts, collagen, cell growth factors, genetic research, animal models,look forward to fully understand the exact mechanism of scar formation, So as to provide scientific theory of prevention and treatment of scar.Goal:To investigate the effect of hydroxycamptothecin (HCPT) on P-Smad3 mRNA and protein expression of human hypertrophic scar fibroblasts and the effect of HCPT on hypertrophic scars in rabbit ears.Methods:(1) Human hypertrophic scar was obtained from patients who were from plastic surgery department. Hypertrophic scar fibroblasts,which were cultured in vitro using tissue culture method, divided into experimental group and blank control group. Blank control group, only to join culture medium; experimental group:the cell with fibroblast culture medium and HCPT (125,250,500ng/ml) intervention 24 hours.The expression of P-Smad3mRNA was measured with RT-PCR, the expression of P-Smad3 protein was measured with Western-blot.(2) Twenty-five adult New Zealand white rabbits were given hypertrophic seals over the ventral surface of the two errs and were randomly divided into the four groups.Group A:NS, Group B:1.25mg/ml, Group C:2.5 mg/ml,Group D:5.0 mg/ml. They were subjected to the drugs once shree days for 4 weeks. Immunohistochemistry(PCNA and Bcl-2)and Hematoxylin-Eosin were used to determinate the scar tissues at 2 and 4 weeks after treatment.. Western-blot(Cx43) was used to determinate the scar tissuses at 4 weeks. Results:(1)The concentrations of(125,250,500ng/ml) HCPT can reduce theP-Smad3 mRNA and protein expression of human hypertrophic scar fibroblasts, and as the concentration increased, the reduction enhanced.The optical density ratio of the P-Smad3mRNA as following:500ng/ml group (157.66±14.39),250 ng/ml group (235.75±20.82),125 ng/ml group (321.64±29.26), blank group (398.23±35.45), there are significant differences in each group (P<0.05).there are also happen in the optical density ratio of theP-Smad3 protein (P<0.05).(2) The proliferative index and the numerical fibroblasts of the scar was significantly diferent betwen the groupC,groupD and groupB,control group(P<0.01).Expression of PCNA and bcl-2 was significantly diferent betwen 5.0 mg/ml group,2.5 mg/ml (C,D) and NS group,groupB at 2 weeks after treatment (P<0.01),and also at 4 weeks.Expression of PCNA and bcl-2 are significantly diferent betwen the 2 weeks and 4 weeks in groups of C,D (P<0.01), but there are no significant diference in groups of A and B (P>0.01). The optical density ratio of the Cx43 protein is significantly diferent between group C (157.46±17.55), groupD (193.68±18.35) and groupA (120.12±12.36),groupB (126.37±13.86) (P<0.01),but there is no significant diference in groups of A and B (P>0.01).Conclusion:(1) HCPT could significantly reduce the expression of p-smad3mRNA and protein of human hypertrophic scar fibroblasts,which has a role in the treatment of hypertrophic scars. (2) HCPT can heal hypertrophic scars of rabbit ear,which may be caused by the decrease of the expression of PCNA,Bcl-2 and the increase of the expression of Cx43,and reduce proliferation and promote apoptosis at fibroblasts, increased gap junction communication between cells.It provide an effective basis for clinical treatment of hypertrophic scars with HCPT. |
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