Wang Weidong, Li Guanglei, Yang Hongming
Dr. Weidong Wang, Chinese PLA Medical School, Beijing, China.
Dr. Guanglei Li, Chinese PLA Medical School, Beijing, China.
Pak J Med Sci. 2018 Jan-Feb;34(1):215-220. doi: 10.12669/pjms.341.13636.
Hypertrophic scar is common in burn patients, but treating result could not meet the expectation of the patients and doctors. We have found that certain concentration level of lipopolysaccharide (LPS) stimulated normal fibroblast cells have statistically similar with fibroblast cells from hypertrophic scar on the phenotype level, and with this work we are trying to figure out which Mitogen-Activated Protein Kinase (MAPK) is affected and how it is affected.
Experiments were conducted in May, 2017 at the first affiliated hospital of the Chinese PLA General Hospital, Beijing, China. We have cultured the cell line of human skin fibroblast cells and randomly divided cells into four groups: control group and three stimulation groups. We have rebuilt the LPS stimulated model of skin fibroblast cells in hypertrophic scar based on our previous work. Experimental groups were stimulated with 0.1ug/mL LPS concentration for 24 hours, 48 hours, and 72 hours, respectively. Then we performed western blot analysis of Erk, p-Erk, JNK, p-JNK, p38 and p-p38. We performed statistical analysis with SPSS 15.0.
LPS can up regulate the MAPK/p38 pathway (p<0.05) and down regulate the MAPK/Erk and MAPK/JNK pathways (p<0.05). The changes of phosphorylated protein are time-related, with longer stimulation duration, significant difference is increased (p<0.05).
MAPKs can play an important role in the formation of hypertrophic scar in the skin. Early intervention through the MAPKs could be a promising target in the prevention of the formation of hypertrophic scar.
肥厚性瘢痕在烧伤患者中很常见,但治疗效果未能达到患者和医生的期望。我们发现,一定浓度水平的脂多糖(LPS)刺激正常成纤维细胞在表型水平上与肥厚性瘢痕来源的成纤维细胞具有统计学相似性,通过这项研究我们试图弄清楚哪种丝裂原活化蛋白激酶(MAPK)受到影响以及如何受到影响。
实验于2017年5月在中国人民解放军总医院第一附属医院(北京)进行。我们培养了人皮肤成纤维细胞系,并将细胞随机分为四组:对照组和三个刺激组。基于我们之前的工作,我们重建了肥厚性瘢痕皮肤成纤维细胞的LPS刺激模型。实验组分别用0.1μg/mL LPS浓度刺激24小时、48小时和72小时。然后我们对Erk、p-Erk、JNK、p-JNK、p38和p-p38进行了蛋白质免疫印迹分析。我们使用SPSS 15.0进行统计分析。
LPS可上调MAPK/p38通路(p<0.05),下调MAPK/Erk和MAPK/JNK通路(p<0.05)。磷酸化蛋白的变化与时间相关,刺激持续时间越长,差异越显著(p<0.05)。
MAPKs在皮肤肥厚性瘢痕的形成中可发挥重要作用。通过MAPKs进行早期干预可能是预防肥厚性瘢痕形成的一个有前景的靶点。