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

doi:10.3969/j.issn.2095-4344.1312

Abstract

Abstract:

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.

Key words: hypertrophic scar, PI3K/AKT signaling pathway, MEK/ERK signaling pathway, animal model, vascular endothelial growth factor

CLC Number:

R446

Li Chenchen1, 2, Liu Baimei1, 2, Liu Yang1, 2, Lü Ying1, 2, Cui Pudong1, 2, An Meiwen1, 2 . Trend similarity of PI3K/AKT and MEK/ERK signaling pathways related protein expression in the hypertrophic scar of Bama pigs and human [J]. Chinese Journal of Tissue Engineering Research, 2019, 23(23): 3673-3679.

Figures/Tables 1

2.1 巴马小型猪皮肤组织和人正常皮肤组织免疫组织化学观察目标蛋白阳性表达信号呈棕黄色颗粒状，并且主要表达于细胞质中。在人体正常皮肤组织中，PI3K/AKT蛋白在表皮层呈阳性表达，在真皮层呈弱阳性表达；MEK蛋白在真皮层中呈阴性表达，ERK蛋白在真皮层中呈弱阳性表达，两者在表皮组织中表达较低；在病理组织中，AKT蛋白呈强阳性表达；MEK蛋白在真皮层中表达较正常组增强，MEK/ERK蛋白在表皮层表达较正常组也明显增强。血管内皮生长因子蛋白在正常皮肤组织及病理组织中均呈阳性表达。在巴马小型猪正常皮肤组织中，血管内皮生长因子蛋白呈弱阳性表达，其余各蛋白的表达定位与人体正常皮肤组织相似；在病理瘢痕组织中，ERK蛋白呈强阳性表达，其余表达情况与人体病理组织大致相同。见图1。 2.2 巴马小型猪模型和人体皮肤组织PI3K/AKT蛋白表达见图2。 2.2.1 巴马小型猪模型 PI3K的平均A值在取皮后30 d和60 d的皮肤组织中略高于0 d组(P30=0.007；P60=0.112)，在取皮后90 d及120 d则逐渐下降回归至0 d组水平(P90 > 0.05；P120 > 0.05)；对于其下游因子AKT，取皮后14 d，平均A值显著低于0 d组(P14=0.01)；取皮后30 d和60 d的组织中平均A值则显著高于0 d组和14 d组(P30=0.000，P60=0.000)，60 d组略高于30 d组；取皮后90 d及120 d，AKT的平均A值变化规律与PI3K一致(P90 > 0.05；P120 > 0.05)。 2.2.2 人体皮肤组织病理增殖期瘢痕组、成熟期瘢痕组PI3K的平均A值均显著高于正常组(P=0.003，P=0.007)，但增殖期瘢痕组、成熟期瘢痕组2组间差异无显著性意义 (P > 0.05)。病理增殖期瘢痕组、成熟期瘢痕组的AKT的平均A值同样高于正常组(P=0.033，P=0.013)。 2.3 巴马小型猪模型和人体皮肤组织MEK/ERK蛋白表达见图2。 2.3.1 巴马小型猪模型 MEK的平均A值在取皮后60 d皮肤组织中高于0 d组及其余各时间点组(P=0.000)，其余几组与0 d组相比较差异无显著性意义(P > 0.05)；ERK的平均A值随时间依次增高，并在取皮后30 d达到最高(P30=0.000)，直到取皮后60 d时ERK的平均A值依然显著高于其余各组(P60=0.003)，但略低于30 d组；取皮后60 d后ERK的平均A值与对照组比较差异无显著性意义(P90 > 0.05；P120 > 0.05)。 2.3.2 人体皮肤组织成熟期瘢痕组MEK/ERK的平均A值明显高于其余2组(PMC=0.000；PEC=0.019)，正常皮肤组、增殖期瘢痕组间2种蛋白的平均A值比较差异无显著性意义(P > 0.05)。 2.4 血管内皮生长因子蛋白表达 2.4.1 巴马小型猪模型血管内皮生长因子的平均A值在取皮后60 d显著高于对照组(P=0.000)，其余时间点平均A值相比差异无显著性意义(P > 0.05)。 2.4.2 人体皮肤组织增殖期瘢痕组血管内皮生长因子的平均A值较正常皮肤组显著升高(P=0.003)，成熟期瘢痕组则显著低于正常皮肤组(P=0.000)。见图2。"

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