瘢痕疙瘩中肿瘤坏死因子α刺激基因-6及相关上下游分子表达水平及相关性的初步探究

Preliminary study on the expression level and correlation of TSG-6 and related upstream and downstream molecules inkeloids

目的:研究瘢痕疙瘩中肿瘤坏死因子α刺激基因(TSG)-6及相关上下游分子的表达水平及相关性。方法:收集78例瘢痕疙瘩样本,另取50例正常皮肤组织样本作为对照。采用免疫组化检测TSG-6的阳性表达率,采用Western blot检测TSG-6、磷酸化磷酸肌醇-3-激酶(p-PI3K)、磷酸化蛋白激酶B(p-AKT)、自杀相关因子(Fas)、Fas配体(FasL)、转化生长因子(TGF)-β1及磷酸化Smad3(p-Smad3)的蛋白表达水平,采用荧光定量PCR法检测组织样本中微小RNA(miR)-23b、miR-181a及miR-376b的表达水平。结果:瘢痕疙瘩组织中TSG-6的阳性表达率、TSG-6、Fas及FasL的蛋白表达水平均低于正常皮肤组织,p-PI3K、pAKT、TGF-β1、p-Smad3、miR-23b、miR-181a及miR-376b的表达水平高于正常皮肤组织(P<0.05);瘢痕疙瘩组织中TSG-6的表达水平与p-PI3K、p-AKT、TGF-β1、p-Smad3、miR-23b、miR-181a及miR-376b的表达水平呈负相关,与Fas及FasL的表达水平呈正相关(P<0.05)。结论:TSG-6表达降低与瘢痕疙瘩形成有关,其机制可能涉及上游miR-23b、miR-181a、miR-376b及下游PI3K/AKT、Fas/FasL、TGF-β1/Smad通路。

Objective:To study the expression level and correlation of tumor necrosis factor alpha stimulated gene-6(TSG-6)and related upstream and downstream molecules in keloids.Methods:A total of 78 cases of keloid samples surgically removed in the First Affiliated Hospital of Hebei North University from January 2020 to October 2021 were collected,and 50 cases of normal skin tissue removed due to open trauma of limbs were taken as control.The positive expression rate of TSG-6 was detected by immunohistochemistry.The protein expression levels of TSG-6,phosphorylated phosphatidylinositol 3 kinase(p-PI3K),phosphorylated protein kinase B(p-AKT),suicide associated factor(Fas),Fas ligand(FasL)and transforming growth factor-β1(TGF-β1),p-Smad3 in tissue samples were detected by Western blot.The expression levels of microRNA(miR)-23b,miR-18la and miR-376b in tissue samples were detected by fluorescence quantitative PCR.Results:The positive expression rate of TSG-6,the protein expression levels of TSC-6,Fas and FasL in keloid tissue were lower than those in normal skin tissue.The expression levels of p-PI3K,p-AKT,TGF-β1,p-Smad3,miR-23b,miR-18la and miR-376b were higher than those in normal skin tissues(P<0.05).The expression level of TSG-6 in keloid tissue was negatively correlated with those of p-PI3K,p-AKT,TGF-β1,p-Smad3,miR-23b,miR-18la and miR-376b,and positively correlated with those of Fas and FasL(P<0.05).Conclusion:The expression of TSG-6 decreased in keloids.The related upstream molecules include miR-23b,miR-18la and miR-376b,and the related downstream molecules include PI3K/Akt,Fas/FasL and TGF-β1/Smad pathway.