粉防己碱对增生性瘢痕成纤维细胞增殖及ERK/NF-κB通路的影响

Effects of tetrandrine on the proliferation of hypertrophic scar fibroblasts in vitro and ERK /NF-κB signaling pathway

目的:观察粉防己碱(tetrandrine,Tet)对增生性瘢痕成纤维细胞增殖及ERK/NF-κB信号通路的影响。方法:培养增生性瘢痕成纤维细胞,四甲基偶氮唑蓝(MTT)比色法、[3H]-TdR掺入率测定不同浓度Tet对细胞增殖的抑制作用;免疫沉淀法纯化蛋白并ERK1/2试剂盒测定ERK活性;Western-blot测定p-ERK1/2、NF-κB(p65)及NF-κB抑制物(I-κBα)的表达。结果:Tet作用下,细胞胞体收缩、变圆,间隙增宽,药物浓度高时,细胞碎裂、浮起。MTT实验、[3H]-TdR掺入率均显示Tet明显抑制细胞增殖,Western-blot显示Tet明显抑制p-ERK1/2及NF-κB(p65)表达及增强I-κBα表达。结论:Tet可抑制增生性瘢痕成纤维细胞增殖,其途径与下调ERK/NF-κB信号通路有关。

Objective To investigate the effects of tetrandrine （Tet） on the proliferation of hypertrophic scar fibroblasts in vitro and ERK/NF-κB signaling pathway. Methods The effects of Tet on proliferation of fibroblasts were observed by MTT assay, [ ^3H ]-TdR incorporation; ERK activity was measured by immuno-precipitation; p-ERK1/2, NF-κB （p65）and I-κBα expressions were assessed by Western blot. Results Fibroblasts exhibited morphological changes： shrinked to round or split into pieces when treated with high concentration of Tet solution. Tet could depress the proliferation of scar fibroblasts. The ERK activity and expressions of p-ERK1/2 and NF-κB （p65） were inhibited and I-κBα expression was increased by different concentrations of Tet. Conclusions Tet can depress the proliferation of fibroblasts in the dose dependent manner. The anti-proliferative effect of Tet may be due to down-regulating ERK/NF-κB signaling pathway.