猪脱细胞真皮基质影响蛋白激酶B/β-连环蛋白信号通路小鼠创面的研究

Effects of porcine acellular dermal matrix on the expression ofβ-catenin and protein kinase B during mouse wounds healing at different time points

目的观察猪脱细胞真皮基质(pADM)在小鼠创面修复过程中对蛋白激酶B(Akt)及β-连环蛋白(β-catenin)表达水平的影响。方法猪皮经脱细胞处理后制成微粒皮,15只C57BL/L小鼠在脊柱左右各制作直径为6 mm的圆形皮肤缺损,左侧以纱布覆盖,常规护理,右侧以pADM覆盖,不作特别处理,按处理方法分为pADM组(实验组)及纱布组(对照组),建模后第1、2、3、4、6周每次取3只小鼠处死,并取创面组织,使用免疫组织化学方法检测Akt及β-catenin在组织中的表达。组间比较采用t检验。结果在模型建立后的第1、2、3、4、6周,实验组各时间点AKT蛋白表达量两两比较差异均有统计学意义(4022.02±652.42、1160.12±225.60、3422.17±675.91、324.15±64.67、27.26±4.49,t1-2=9.458、t2-3=-4.839、t3-4=8.776、t4-6=7.436,P<0.05)。同时对照组蛋白表达在各时间点间有统计学意义(1110.98±219.14、337.13±61.37、118.80±18.08、834.98±64.19、395.70±76.79,t1-2=8.802、t2-3=4.951、t3-4=-19.584,t4-6=6.835,P<0.05)。第2、3、4、6周时实验组(1160.12±225.60、3422.17±675.91、324.15±64.67、27.26±4.49)及对照组(337.13±61.37、118.80±18.08、834.98±64.19、395.70±76.79)的蛋白表达差异有统计学意义(t2=8.574、t3=8.694、t4=-10.183、t6=-7.880,P<0.05),其他组差异无统计学意义(4022.02±652.42、1160.12±225.60,t1=-0.363,P>0.05)。实验组各时间点β-catenin蛋白表达量两两比较差异均有统计学意义(3646.32±687.04、8447.29±1181.79、9488.65±1439.52、6158.74±1083.52,t1-2=-6.626、t3-4=12.101,P<0.05),对照组蛋白表达在各时间点间有统计学意义(4364.09±811.29、2187.85±424.84、2752.27±500.01、7522.95±1109.30、2592.14±474.23,t1-2=-4.334、t3-4=-5.594、t4-6=12.154,P<0.05),实验组(8447.29±1181.79、9488.65±1439.52)及对照组(2187.85±424.84、2752.27±500.01)两组间的β-catenin的蛋白表达在第2、3周时差异有统计学意义(t2=9.351、t3=-9.225,P<0.05),其他组差异无统计学意义(3646.32±687.04、6158.74±1083.52、5487.60±1068.96;4364.09±811.29、7522.95±1109.30、2592.14±474.23,t1=0.377、t4=0.386、t6=0.051,P>0.05)。实验组及对照组两组间的Akt及β-catenin蛋白在创面组织中的表达,均于第2周开始升高,于第3周达高峰,具有相似的变化趋势。结论pADM可能是通过影响Akt/β-catenin信号通路的表达从而影响创面愈合。

Objective To investigate the effect of porcine acellular dermal matrix(pADM)on the expression ofβ-catenin and protein kinase B(Akt)during mouse wounds healing at different time points after skin injury.Methods The skin injury models were established in 15 C57BL/6 mice.The spine was set as the midline,and on its both sides a full-thickness skin defect wound with 6 mm diameter was made.The left wound was covered with gauze(control group),and the right wound was covered with particulate pADM(pADM group),and the tissues of wound edge were collected at the 1st,2nd,3rd,4th,and 6th week after the establishment of the model.The expression ofβ-catenin and Akt proteins was detected by immunohistochemical staining,and that of the two kind proteins in tissues and cells was observed pathologically.T test was used for comparison between groups.Results Immunohistochemical detection of protein expression in mouse wound tissue showed that the protein expression levels ofβ-catenin and Akt were different in the two groups,and all of them had statistical significance(respectively).For the expression of Akt protein,there was significant difference in pADM between the two groups,significant differences between 1st week and 2nd week,2nd week and 3rd week,3rd week and 4th week,4th week and 6th week(4022.02±652.42,1160.12±225.60,3422.17±675.91,324.15±64.67,27.26±4.49,t1-2=9.458,t2-3=-4.839,t3-4=8.776,t4-6=7.436,P<0.05),no significant differences between the other groups,in control group,significant differences between 1st week and 2nd week,2nd week and 3rd week,3rd week and 4th week,4th week and 6th week(1110.98±219.14,337.13±61.37,118.80±18.08,834.98±64.19,395.70±76.79,t1-2=8.802,t2-3=4.951,t3-4=-19.584,t4-6=6.835,P<0.05),significant difference existed at the 2nd,3rd and 6th week between pADM group(1160.12±225.60,3422.17±675.91,324.15±64.67,27.26±4.49)and control group(337.13±61.37,118.80±18.08,834.98±64.19,395.70±76.79,t2=8.574,t3=8.694,t4=-10.183,t6=-7.880,P<0.05),but no significant difference was found at the 1st week.A similar change trend of Akt andβ-catenin existed in pADM group,both of which began to increase at the 2nd week and reached the peak at the 3rd week,when the expression difference between the two factors was the largest in pADM group and control group,and decreased from the 3rd week.For the expression ofβ-catenin protein,there was significant difference in pADM between the two groups,significant differences between 1st week and 2nd week,3rd week and 4th week(3646.32±687.04,8447.29±1181.79,9488.65±1439.52,6158.74±1083.52,t1-2=-6.626,t3-4=12.101,P<0.05),no significant differences between the other groups,in control group,significant differences between 1st week and 2nd week,3rd week and 4th week,4th week and 6th week(4364.09±811.29,2187.85±424.84,2752.27±500.01,7522.95±1109.30,2592.14±474.23,t1-2=-4.334,t3-4=-5.594,t4-6=12.154,P<0.05),significant difference existed at 2nd,3rd and 6th week between pADM group(8447.29±1181.79,9488.65±1439.52)and control group(2187.85±424.84,2752.27±500.01,t2=9.351,t3=9.225,P<0.05),and no significant difference was found at the rest period.Histochemical observation showed thatβ-catenin and Akt were expressed in wound tissues.Conclusion Akt/β-catenin signal pathway is involved in wound healing.The pADM can promote wound healing probably by promoting the expression of Akt/β-catenin signal pathway.