靶向p38丝裂素活化蛋白激酶对兔唇裂术后瘢痕增生的影响  

Effect of targeting p38MAPK on scar hyperplasia after rabbit cleft lip repair in vivo

作　　者：葛盈盈 岳金[1,2] 薛令法[1,2] 许尧祥[1,2] 赵浩然 崔明雪 肖文林[1,2] Yingying Ge;Jin Yue;Lingfa Xue;Yaoxiang Xu;Haoran Zhao;Mingxue Cui;Wenlin Xiao(Department of the Affiliated Hospital of Qingdao University,Qingdao 266555,China;School of Stomatology of Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学附属医院口腔颌面外科,青岛266555 [2]青岛大学口腔医学院,青岛266071

出　　处：《中华口腔医学研究杂志（电子版）》2023年第1期37-44,共8页Chinese Journal of Stomatological Research(Electronic Edition)

基　　金：山东省自然科学基金(ZR2015HM022)。

摘　　要：目的研究运用p38丝裂素活化蛋白激酶(p38MAPK)基因重组腺病毒靶向敲低目的基因的表达,通过检测p38MAPK信号通路受到抑制后不同时间段兔上唇瘢痕增生受到的影响,来探讨疗效最佳的基因治疗时间。方法对90只2.0~2.5 kg的上唇裂新西兰白兔进行唇裂修复术,选取术后第0、1、2周对其瘢痕正中注射重组病毒,将第3周的瘢痕组织制成标本。通过使用蛋白免疫印迹法(Western blot)、实时荧光定量反转录聚合酶链反应(RT-PCR)、免疫组织化学染色等方法分别定性、定量检测p38MAPK及瘢痕形成相关因子与Smad蛋白和mRNA的相对表达水平。使用SPSS 24.0软件对实验结果进行统计学分析。结果相较于术后第0、2周,术后第1周瘢痕组织中ColⅢ(1.373±0.073,F=8.027,P=0.002)、MMP1(1.715±0.028,F=9.262,P=0.001)表达明显升高,ColⅠ(0.424±0.015,F=7.794,P=0.003)和TIMP1(0.464±0.025,F=6.196,P=0.007)表达明显降低,差异均有统计学意义。术后第1周的Smad2蛋白表达水平与mRNA表达水平降低(F=14.123,P=0.029),Smad3蛋白表达水平与mRNA表达水平降低(F=3.796,P=0.037),与其他组相比差异均有统计学意义。结论抑制p38MAPK表达可能通过Smad依赖型信号通路发挥抑制瘢痕增生的作用,兔上唇唇裂术后第1周瘢痕形成过程中靶向敲低p38MAPK对瘢痕增生影响最大。Objective The recombinant adenovirus of p38 mitogen-activated protein kinases(p38MAPK)gene was used to knock down the expression of target genes.By detecting the influence of p38MAPK signal pathway obstruction on upper lip scar hyperplasia in different time periods,the optimal therapeutic time of gene therapy was determined.Methods A total of ninety New Zealand white rabbits with cleft upper lip of 2.0-2.5 kg were treated with cleft lip repair.Recombinant virus was injected into the scar center at week 0,1 and 2 after surgery,and the scar tissue was made into specimens at week 3 after surgery.Western blot,Real-time fluorescence quantitative RT-PCR and immunohistochemical staining were used to quantitatively and qualitatively detect the relative expression levels of proteins and mRNA of p38MAPK,scar formation related factors and Smad.The results of Western blot and RT-PCR were analyzed with software(SPSS v.24.0).Results The results of Western blot,RT-PCR and immunohistochemical staining showed that the expressions of ColⅢ(1.373±0.073,F=8.027,P=0.002)and MMP1(1.715±0.028,F=9.262,P=0.001)in scar tissue of the injection group at week 1 after surgery were significantly higher than those at week 0 and 2.The expressions of ColⅠ(0.424±0.015,F=7.794,P=0.003)and TIMP1(0.464±0.025,F=6.196,P=0.007)significantly decreased.The expression level of Smad protein and mRNA in the injection group at 1 week after surgery decreased,and the results were statistically significant compared with other groups(F_(p-smad2)=14.123,P_(p-smad2)=0.029;F_(p-smad3)=3.796,P_(p-smad3)=0.037).Conclusions Inhibition of p38MAPK expression may play a role in inhibiting scar hyperplasia through the Smad dependent signaling pathway.Targeted p38MAPK knockdown had the greatest effect on scar hyperplasia at one week after cleft-lip surgery in rabbit upper lip.

关 键 词：P38丝裂素活化蛋白激酶 基因沉默 腺病毒 增生性瘢痕 基因治疗 

分 类 号：R78[医药卫生—口腔医学]