白芷活性提取物对成纤维细胞生物学特性的影响  

Effects of Angefica dahurica extract on biological behavior of dermal fibroblasts

作　　者：白晓智[1] 胡大海[1] 王耘川[1] 刘佳琦[1] 石继红[1] 汤朝武[1] 

机构地区：[1]第四军医大学西京医院全军烧伤中心,西安710032

出　　处：《中华外科杂志》2012年第4期357-360,共4页Chinese Journal of Surgery

基　　金：陕西省“13115”科技创新工程重大科技专项资助项目（2009ZDKG-87）

摘　　要：目的体外观察白芷活性提取物对人皮肤成纤维细胞（Fh）生物学特性的影响，探讨其促进创面愈合的可能机制。方法分离培养人正常Fb，按照随机数字表法分白芷活性提取物不同浓度组，以体积分数0．25％的新生牛血清DMEM为对照组。采用噻唑蓝比色法（MTT）检测不同浓度白芷活性提取物对体外培养的Fb增殖作用的影响。通过流式细胞仪、real-time PCR分别检测白芷活性提取物最适浓度培养条件下Fb的细胞周期变化以及Ⅰ、Ⅲ型胶原mRNA的表达情况。结果白芷活性提取物在5×10^-4-5×10^-2g／L浓度范围内可显著促进Fb增殖，呈剂量依赖性，不同浓度白芷活性提物培养液组测得吸光度（A）值分别为0．346±0．055、0．387±0．043和0．310±0．026，与对照组（0．2734-0．042）比较，差异有统计学意义（t=3．36、5．79和2．49，P〈0．05），其中在5×10^-3g／L浓度时测得A值最高，促增殖作用最为显著，为白芷活性提取物促进Fh增殖的最适合浓度。此浓度条件下白芷活性提取物可明显促进Fh通过G1／S及S／G2期限制点，S期（26．6±2．6）及G2／M期（7．4±1．3）细胞与对照组相比明显增多，G0／G1期细胞（66．3±4．5）与对照组相比明显减少（t=11．2、5．69、2．44，P〈0．05）。5×10^-3g/L白芷活性提取物组与对照组比较I、Ⅲ型胶原mRNA表达明显上调（分别为1．61±0．26比1．00±0．16和3．36±0．40比1．00±0．14），差异有统计学意义（t=6．69、7．64，P〈0．叭）。结论白芷活性提取物能显著促进Fh增殖，加快Fb细胞周期进程，同时可上调Ⅰ、Ⅲ型胶原mRNA的表达，可能与白芷促进创面愈合的机制有关。Objective To observe the effects of Angelica dahurica extracts on the biological characteristics of human dermal fibroblasts in vitro and to preliminary explore its possible therapeutic mechanism for wound healing. Methods The optimal concentration of Angelica dahurica extracts was identified by analysing of proliferation activity of human normal fibroblasts （Fb） that treated with different concentration of Angelica dahurica extracts through thiazole blue （MTF） colorimetric assay. Cell cycle, collagen Ⅰ and collagen Ⅲ mRNA levels of the optimal Angelica dahurica extracts treated Fb were detected by flow cytometry （FCM） and real-time PCR techniques. Results At concentrations of 5×10^-4 to 5×10^-2 g/L, the Angelica dahurica extracts significantly enhanced the proliferation of Fb. The most significant concentration was 5×10^-3 g/L （t=5.79, P 〈0. 01 ）, at which an increased percentage of G1 to S and S to G2 phase cells （t = 11.2, 5.69, 2. 44, P 〈 0.05） as well as an increased level of collagen Ⅰ （ 1.61±0. 26 vs. 1.00±0. 16） and collagen Ⅲ mRNA （3.36±0. 40 vs. 1.00±0. 14） were obtained compared to the control group （ t = 6.69, 7.64, P 〈 0.01 ）. Conclusions Angelica daharica extracts can notably promote the proliferation of Fb and accelerating the cell cycle of Fb as well as up-regulating the expression of collagen Ⅰ and collagen Ⅲ, which may enhance the process of wound healing.

关 键 词：白芷 成纤维细胞 伤口愈合 胶原Ⅰ型 胶原Ⅲ型 

分 类 号：R285[医药卫生—中药学]