Gragnani Alfredo, Müller Bruno Rafael, Silva Ismael Dale Contrim Guerreiro da, Noronha Samuel Marcos Ribeiro de, Ferreira Lydia Masako
Division of Plastic Surgery, Department of Surgery, Federal University of Sao Paulo, Paulista School of Medicine, Brazil.
Acta Cir Bras. 2013 Aug;28(8):551-8. doi: 10.1590/s0102-86502013000800001.
To evaluate the gene expression of KGF, TNF-alpha and IL-1 beta in skin fibroblasts and keratinocytes cultured from burned patients.
Three patients with large burns and three patients with small burns, as well as two controls, were included. The cell culture was initiated by the enzymatic method. After extraction and purification of mRNA, qPCR was used to assess the gene expression of KGF, TNF-alpha and IL-1 beta.
The expression of KGF was increased on average 220-fold in large burns and 33.33-fold in small burns in fibroblasts, and 11.2-fold in large burns and 3.45-fold in small burns in keratinocytes compared to healthy patients (p<0.05). Expression of TNF-alpha was not observed. IL-1 beta is down-regulated in fibroblasts of burned patients, and much more repressed in small burns (687-fold, p<0.05). In keratinocytes, the repression of IL-1 beta expression occurs in patients with small burns (28-fold), while patients with large burns express this gene intensively (15-fold).
The study showed a quantitative pattern in the expression of KGF gene, which is more expressed according to the size of the burn. TNF-alpha was not expressed. A qualitative pattern in the expression of IL-1 beta gene was demonstrated.
评估烧伤患者培养的皮肤成纤维细胞和角质形成细胞中KGF、TNF-α和IL-1β的基因表达。
纳入3例大面积烧伤患者、3例小面积烧伤患者以及2例对照。采用酶法进行细胞培养。mRNA提取和纯化后,用qPCR评估KGF、TNF-α和IL-1β的基因表达。
与健康患者相比,成纤维细胞中KGF的表达在大面积烧伤中平均增加220倍,在小面积烧伤中增加33.33倍;角质形成细胞中KGF的表达在大面积烧伤中增加11.2倍,在小面积烧伤中增加3.45倍(p<0.05)。未观察到TNF-α的表达。烧伤患者成纤维细胞中IL-1β表达下调,在小面积烧伤中更受抑制(687倍,p<0.05)。在角质形成细胞中,小面积烧伤患者中IL-1β表达受到抑制(28倍),而大面积烧伤患者中该基因表达强烈(15倍)。
该研究显示了KGF基因表达的定量模式,其表达根据烧伤面积而增加。未表达TNF-α。证明了IL-1β基因表达的定性模式。