小鼠皮肤切创愈合过程中血清游离DNA的改变

Changes of cf-DNA during wound healing in mouse skin

①目的检测皮肤切创愈合过程中血清游离DNA(cf-DNA)变化的时间规律性。②方法将健康小鼠40只随机分为8组:对照组、0.5小时组、1小时组、2小时组、24小时组、48小时组、4天组和16天组,每组5只小鼠。除对照组外,其余各组建立皮肤切创模型。通过内眦静脉采血后提取血清cf-DNA,随机选取3份DNA样本验证性扩增保守基因(β-actin)。采用荧光核酸蛋白测定仪对cf-DNA进行检测与定量,SPSS13.0统计分析软件对各组结果进行分析处理。③结果3份DNA样本均检测到清晰条带,且与β-actin阳性样本位置一致。与对照组相比,0.5小时、1小时和16天时相组cf-DNA含量无明显差异(P>0.05)。2小时、24小时、48小时和4天时相组血清cf-DNA含量明显升高(P<0.05),其中48小时组最高。④结论在皮肤创伤愈合过程中血清cf-DNA浓度变化具有一定的规律性及时间相关性,有望成为推断创口形成时间的生物学指标。

Objective To investigate the cf-DNA in serum and its time-dependent changes during the skin incised wound healing.Methods A total of 40 healthy mice were randomly divided into control group,0.5h group,1h group,2h group,24h group,48h group,4d group and 16d group,5 mice in each group.Except foRthe control group,the skin incision models of mice were established in the otheRgroups.Serum cf-DNA was extracted from the inneRcanthus vein afteRblood collection,three DNA samples were randomly selected to verify the amplification of conservative gene beta-actin.Serum cf-DNA was detected and quantified by fluorescent nucleic acid protein analyzer.The results of each group were analyzed and processed by SPSS statistical analysis software.Results CleaRbands were detected in all three DNA samples,and the location was consistent with that of beta-actin positive samples.Compared with the control group,there was no significant difference in cf-DNA concentration between 0.5h,1h and 16d phase groups( P > 0.05 ).Serum cf-DNA concentration increased significantly in 2h,24h,48h and 4d phase groups( P < 0.05 ).The highest cf-DNA concentration was found in 48h phase group.Conclusion The concentration of cf-DNA inserum is time-dependently expressed in skin wound healing,which can be a useful markeRfoRwound age determination.