没药提取物对人成纤维细胞增殖与胶原mRNA表达影响的实验研究  

Effects of myrrh extract on proliferation and collagen mRNA expression of human fibroblasts in vitro

作　　者：白晓智[1] 胡大海[1] 白丽[1] 刘洋[1] 苏映军[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,烧伤与皮肤外科,西安710032

出　　处：《中华烧伤杂志》2012年第2期130-133,共4页Chinese Journal of Burns

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

摘　　要：目的观察没药提取物对人皮肤Fh生物学特性的影响，探讨其促进创面愈合的可能机制。 方法从人正常包皮组织中分离并培养Fb，取第3-5代细胞用于实验。（1）将Fh接种至96孔板，按随机数字表法分为对照组，1×10-4、1×10 1、1×10-3、1×10-21、1、10、1×102 g/L没药水提取物组以及上述7种浓度的没药醇提取物组。对照组用含体积分数0. 25%小牛血清的DMEM培养液（简称低浓度血清培养液）培养，各浓度没药水及没药醇提取物组分别用含相应终浓度2种没药提取物的低浓度血清培养液培养。培养48 h，用倒置相差显微镜观察细胞形态，噻唑蓝法测定各组Fh增殖活性（以吸光度值表示）。（2）将Fh分别接种于培养瓶和培养皿中，均按随机数字表法分为2组：1 g/L没药水提取物组，用含终浓度1 g/L没药水提取物的低浓度血清培养液培养；对照组，采用低浓度血清培养液培养。培养72 h，分别采用流式细胞仪、实时荧光定量PCR法检测Fb周期与I、Ⅲ型胶原mRNA的表达。对数据进行LSD-t检验。 结果 （1）各组细胞均呈长梭形生长，1 g/L没药水提取物组细胞融合较对照组更显著。1×10-3、1×10-2、1×10-11，1、10 g/L没药水提取物组Fb吸光度值分别为0. 378±0.032、0.402±0.007、0.390±0.038、0.453±0.036、0.390±0.037，均高于对照组的0. 332±0.044，t值分别为2.24、2.93、2.69、5.73、2.71，P值均小于0.05。1×10-3、1×10-2 g/L没药水提取物组吸光度值分别为0.312±0.048、0.154±0.009，前者与对照组比较，差异 无统计学意义（t=2.84，P〉0.05）；后者显著低于对照组（t=7. 17，P〈0.05）。1×10。3、1×10。1、1、10、1×102 g/L没药醇提取物组Fh吸光度值显著低于对照组（z值为2.30 24. 79，P值均小于0. 05）。（2）1 g/L没药水提取物组CO/G1期细胞百分比为（74.3±6.3）%，明显少于对照组的（82.2 ±7.9）010，z=6.77，P〈0.05；S�Objective To observe the effects of myrrh extract on biological characteristics of human dermal fibroblasts （Fb） , and to explore its possible mechanisms in promoting wound healing. Methods Normal Fb was isolated from human foreskin tissue and cuhured in vitro. The third to fifth passages of Fb were used in the experiment. （ 1 ） Fb were planted onto 96-well plate and divided into control group, and 1 x10-, 1 x10 3, 1 ×10^-2, 1 ×10^ -1, 1, 10, 1 ×10^2 g/L myrrh water extract groups and myrrh ethanol extract groups according to the random number table. Fb in control group were cultured with DMEM medium containing 0.25% calf serum （briefly called low-concentration serum medium）, and those in various concentrations of myrrh water extract and myrrh ethanol extract groups respectively with low-concentration serum medium containing corresponding concentration of 2 kinds of myrrh extract. After being cultured for 48 h, cell morphology was observed with inverted-phase contrast microscope, and Fb proliferation activity （ denoted as absorbance value） was determined with MTT method. （2） Fb were respectively planted into flasks and dishes and divided into two groups according to the random number table. Fb in control group were cultured with low-concentration serum medium, and that in 1 g/L myrrh water extract group with low-concentration serum medium containing 1 g/L myrrh water extract. After being cultured for 72 h, Fb cell cycle and the type Ⅰ and Ⅲ collagen mRNA expression were respectively determined by flow cytometry and real-time fluorescent quantitative PCR. Data were processed with LSD- t test. Results （ 1 ） Fb in all groups grew in long-spindle shape, but the cell fusion was much obvious in 1 g/L myrrh water extract group than in control group. Fb absorbanee value in 1 ×10-3, 1 ×10-2, 1 ×10-1, 1, 10 g/L myrrh water extract groups was respectively 0. 378 ± 0. 032, 0. 402 ± 0. 007, 0. 390 ± 0. 038, 0. 453 ± 0. 036, 0. 390 ± 0. 037, all higher than that in control group （0. 33

关 键 词：没药属 成纤维细胞 细胞增殖 胶原 伤口愈合 

分 类 号：R285.5[医药卫生—中药学]