摘要
目的观察鸦胆子油乳（brucea javanica oil emulsion，BJOE）对增生性瘢痕成纤维细胞（hypertrophic scar fibroblasts，HSFs）增殖、胶原合成和ERK1/2 MAPK信号通路的影响。方法收集人来源增生性瘢痕组织，行HSFs体外培养及鉴定；终浓度为0.5、1、5、10、20、50 mg/ml浓度的BJOE作用HSFs 24 h后，计算出24 h半数抑制浓度（IC50）。以近IC50浓度的BJOE作用HSFs 24、48、72 h为实验组，对照组为常规培养的HSFs，CCK-8法检测细胞的增殖活力，流式细胞术（FCM）进行细胞的周期检测。观察近IC50浓度的BJOE作用HSFs 24 h后细胞的形态学变化。Western blot法检测Ⅰ型胶原、波形蛋白、细胞外信号调节激酶1/2（ERK1/2）及其磷酸化蛋白p-ERK1/2的表达。结果BJOE的IC50为1.176 mg/ml，故选择1 mg/ml浓度进行后续实验。以1 mg/ml浓度的BJOE作用24、48、72 h后，BJOE对HSFs的抑制率分别为（53.13±2.40）%、（75.07±2.67）%、（88.65±0.32）%，差异具有统计学意义（P〈0.05）。BJOE作用下HSFs数目明显减少，细胞间隔增宽，细胞突起明显缩短，甚至消失，细胞变圆。实验组的S期细胞百分数逐渐减少。HSFs中Ⅰ型胶原、波形蛋白和p-ERK1/2的表达明显受到抑制（P〈0.05），而ERK1/2的表达无明显变化（P〉0.05）。结论鸦胆子油乳对HSFs的增殖及Ⅰ型胶原的合成具有抑制作用，其机制与抑制ERK1/2磷酸化及波形蛋白的表达有关。
Objective To investigate the effect of brucea javanica oil emulsion （BJOE） on the proliferation, collagen synthesis and ERK1/2 MAPK singaling pathway of hypertrophic scar fibroblasts （ HSFs）. Methods The human hypertrophic scar tissues were collected, and HSFs were isolated, cultured and identified in vitro. Cells were treated with BJOE under the concentration of 0.5, 1 , 5, 10, 20 and 50 mg/ml for 24 h respectively; the half inhibitory concentration （IC50） of 24 h was calculated. The experimental groups were treated for 24,48,72 h by B JOE at the IC50, the control group was conventionally cultured without B JOE. Cell proliferation was detected by CCK-8 assay and cell cycle was determined by flow cytometry （ FCM ）. The morphological changes of cells treated with BJOE at a concentration of IC50 for 24 h were observed. The protein expression levels of collagen I , vimentin, ERKl/2 and p-ERK1/2 were measured by Western blot. Results The IC50 of BJOE was 1. 176 mg/ml, so the concentration of 1 mg/ml was selected for the subsequent experiment. After 24,48,72 h treated with B JOE at a concentration of 1 mg/ml, the inhibition rates of BJOE on HSFs were （53.13 ± 2.40） % , （75.07 ±2.67 ） % and （ 88.65±0.32 ） % , respectively. The difference was significant （ P 〈 0.05 ）. Under the effect of B JOE, the number of HSFs significantly decreased, the interval between cells widened, cell protrusions were significantly shortened or even disappeared, and the cells became round. The percentage of cells decreased in S phase. Expression of type I collagen, vimentin and p-ERK1/2 in HSF was significantly inhibited （P 〈 0. 05 ） , while there was no significant difference in the expression of ERK1/2 （ P 〉 0.05 ）. Conclusions BJOE inhibites the proliferation of HSF and the synthesis of type I collagen, and its mechanism is related to the inhibition of ERK1/2 phosphorylation and vimentin expression.
作者
刘松健
章宏伟
张春丽
Liu Songjian;Zhang Hongwei;Zhang Chunli(Department of Burn and Plastic Surgery, First Affiliated Hospital of Nanjing Medical University, Nanjing 210029, China)
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2018年第5期397-402,共6页
Chinese Journal of Plastic Surgery
