抑制作用及机制研究芍药苷对增殖性瘢痕成纤维细胞增殖

摘要

目的 探索芍药苷对增殖性瘢痕(HS)成纤维细胞增殖抑制作用及机制研究.方法 0(对照组), 200,400,800 μmol/L的芍药苷作用HS成纤维细胞24、48、72 h后,MTT法检测细胞活力,Hoechst染色法检测细胞凋亡,流式细胞术检测细胞周期,酶联免疫吸附试验(ELISA)检测细胞Ⅰ型胶原(COL Ⅰ)和Ⅲ型胶原(COL Ⅲ)含量,Western blot检测转化生长因子-β(TGF-β)/Smad信号通路相关蛋白及基质金属蛋白酶1(MMP1)、MMP13表达.结果 200,400,800 μmol/L芍药苷能显著降低HS成纤维细胞活力(P< 0.01),使细胞核发白,皱染,使细胞周期阻滞在G1期(P< 0.01),能显著降低细胞中COLⅠ、COLⅢ含量(P< 0.01),下调MMP1、MMP13、TGF-β1、p-Smad2及p-Smad3表达(P< 0.01).结论 芍药苷能明显抑制HS成纤维细胞增殖及胶原合成,可能是通过抑制TGF-β1/Smad信号通路实现的.%Objective To explore the inhibitory effect and related mechanism of paeoniflorin on proliferation of hypertrophic scar(HS)fibroblasts. Methods HS fibroblasts were cultured with 0(control),200,400,800 μmol/L of paeoniflorin for 24,48,and 72 h. Cell viability was detected by MTT assay. Cell apoptosis was detected using Hoechst staining. Cell cycle was measured by flow cytometry. The levels of type I collagen(COL I)and type III collagen(COL III)were detected by enzyme linked immunosorbent assay(ELISA). The expressions of transforming growth factor-β (TGF-β)/Smad signaling pathway related proteins, as well as matrix metalloproteinase 1(MMP1)and MMP13 were detected by Western blot. Results 200, 400, 800 μmol/L of paeoniflorin reduced the cell viability of HS fibroblasts significantly(P < 0.01), with the nuclei turning pale and shrunken, and caused cell cycle arrest at G1 phase(P <0.01). Moreover, the levels of COL I and COL III in the cells were decreased significantly(P < 0.01), and the expressions of MMP1, MMP13, TGF-β1, p-Smad2 and p-Smad3 were down-regulated significantly(P < 0.01). Conclusions Paeoniflorin can obviously inhibit the proliferation and collagen synthesis of hypertrophic scar fibroblasts,probably through inhibition of the TGF-β1/Smad signaling pathway.