巴马小型猪与人体瘢痕组织中PI3K/AKT和MEK/ERK信号通路相关蛋白的表达趋势相似性研究

Trend similarity of PI3K/AKT and MEK/ERK signaling pathways related protein expression in the hypertrophic scar of Bama pigs and human

背景:增生性瘢痕的发生原因、发生时间、发生的对象以及治疗方式是目前研究的重点与难点。研究表明,磷脂酰肌醇-3激酶(phosphatidylinositol 3-kinase,PI3K)/蛋白激酶B(protein kinase B,AKT)和丝氨酸/苏氨酸蛋白激酶(mitogen-activated protein kinase,MEK)/细胞外信号调节激酶(extracellular signal-regulated kinase1/2,ERK1/2)信号通路及其下游因子在细胞增殖与凋亡、侵袭与迁移、血管生成以及致纤维化等过程中发挥了重要作用。目的:分析巴马小型猪瘢痕动物模型与人体的瘢痕组织中PI3K/AKT及MEK/ERK信号通路相关蛋白的表达,探讨巴马小型猪作为动物瘢痕模型的可行性。方法:人皮肤标本来源于太钢总医院山西省烧伤救治中心整形科;猪皮肤标本来源于课题组前期建立的巴马小型猪瘢痕动物模型。所有患者均了解试验的过程并自愿参加。研究方案由太钢总医院伦理委员会讨论通过及并经动物实验伦理委员会批准。将人皮肤标本分为正常皮肤组、增殖期瘢痕组、成熟期瘢痕组;巴马小型猪瘢痕模型根据伤后取材时间分为0d组(对照组)、14d组、30d组、60d组、90d组和120d组。采用免疫组织化学法检测人体及猪模型不同皮肤组织中PI3K、AKT、MEK、ERK、血管内皮生长因子蛋白的表达。结果与结论:①PI3K、AKT、MEK、ERK、血管内皮生长因子蛋白在瘢痕动物模型的正常皮肤组织中呈阴性表达,在病理性瘢痕组织中呈阳性表达,且平均吸光度与正常皮肤组织比较,差异均有显著性意义(P<0.01);②5种蛋白的表达在伤后30-60d达到峰值,随后逐渐降低,回归正常皮肤中的表达水平,与人体皮肤瘢痕组织中相关蛋白的表达趋势相似;③在巴马小型猪皮肤与人体皮肤组织中PI3K/AKT和MEK/ERK信号通路相关蛋白的表达趋势具有相似性,为巴马小型猪是瘢痕形成机制研究的理想动物模型提供了又一个证据。

BACKGROUND:Etiology,time,subjects and treatment methods of hypertrophic scar are issues of concern and difficulties.Phosphatidylinositol 3-kinase(PI3K)/protein kinase B(AKT)and mitogen-activated protein kinase(MEK)/extracellular signal-regulated kinase1/2(ERK1/2)signaling pathways and downstream factors have been shown to play an important role in cell proliferation and apoptosis,invasion and migration,angiogenesis and fibrosis.OBJECTIVE:To explore the feasibility of Bama pig as an animal model of hypertrophic scar by analyzing the expression and distribution of PI3K/AKT and MEK/ERK signaling pathways related proteins in the hypertrophic scar model of human and Bama pigs.METHODS:Human samples were from the Department of Plastic of Tisco General Hospital Shanxi Burn Treatment Center,and the pig skin samples were form the previous Bama pig model of scar established by our group.All patients signed the informed consents,and the study was approved by the Laboratory Animal Ethics Committee of Tisco General Hospital.The human samples were divided into normal skin,hypertrophic scar and mature scar groups.The Bama pig models of scar were divided into 0 day(control group),14-,30-,60-,90-,and 120-day groups.Immunohistochemistry was used to detect the expressions of PI3K,AKT,MEK,ERK and vascular endothelial growth factor proteins in the human and pig skin tissues.RESULTS AND CONCLUSION:(1)The expression of PI3K,AKT,MEK,ERK,and VEGF protein in the normal skin tissue of animal model of scar was negative,and positive in pathological scar.The average absorbance value showed significant difference compared with the normal skin tissues(P<0.01).(2)The expression of the five proteins peaked at 30-60 days after injury,and then decreased gradually to the normal level,which was similar to the trend in human skin scar tissue.(3)To conclude,the protein expression level related to PI3K/AKT and MEK/ERK signaling pathways in Bama pig skin is similar to human skin,which provides evidence for using Bama pig as animal model to study the mechanism of scar formation.