氧化苦参碱抗角膜瘢痕作用机制的研究

The Mechanism of Corneal Scar Inhibition of Oxymatrine

目的:研究不同等级浓度的氧化苦参碱(oxymatrine,OM)对兔角膜成纤维细胞增殖和转化生长因子-β_1(transforming growth factor-β_1,TGF-β_1)及α-平滑肌肌动蛋白(α-smooth muscle actin,α-SMA)mRNA表达的影响,从而探讨OM抑制角膜瘢痕增生的机制。方法:选用试验用新西兰大白兔采集兔角膜成纤维细胞并进行培养,运用甲基噻唑基四唑(methyl thiazolyl tetrazolium,MTT)比色法观察不同OM浓度(20、40、60、80、100μg/mL)对兔角膜成纤维细胞增殖的影响,运用RT-PCR技术研究不同浓度OM对兔角膜成纤维细胞TGF-β_1 mRNA及α-SMA mRNA表达的影响。结果:MTT法检测结果显示,加入不同浓度OM的实验组与对照组(0.631±0.004)相比较,实验组兔角膜成纤维细胞增殖均显著受抑制(OD值分别为0.568±0.002、0.465±0.002、0.331±0.004、0.236±0.003、0.105±0.002),差异有统计学意义(P<0.01):不同浓度(20、40、60、80、100μg/mL)OM均可抑制兔角膜成纤维细胞TGF-β_1mRNA的表达,且随着浓度的增大,其抑制作用增强;从正常兔角膜成纤维细胞中未检测到α-SMA mRNA的表达。结论:OM对体外培养的兔角膜成纤维细胞增殖及TGF-β_1mRNA的表达具有抑制作用,并随着OM浓度的增加,其抑制作用也增强。

Objective： To study the influence of oxymatrine （OM） in different concentrations on fibroblast proliferation and expression of transforming growth factor-β1（TGF-β1） and a-smooth muscle actin （α-SMA） mRNA in rabbits＇ cornea, thereby to investigate the mechanism that OM inhibits corneal scar hyperplasia. Methods： New Zealand rabbits were used to collect corneal flbroblasts and culture in vitro, observed the effects of different OM concentrations （20, 40, 60, 80, 100pg/mL） on fibroblast proliferation in rabbits＇ cornea by methyl thiazolyl tetrazolium （MTT） colorimetric assay, and studied the influence of different OM concentrations on the expression of TGF-β, mRNA and α-SMA mRNA in rabbits＇ corneal fibroblasts. Results： The MTT detection results showed that, compared with control group （0.631 ±0.004）, the corneal fibroblasts proliferation of experimental groups which were in different OM concentrations had significantly inhibited （which OD were 0.568±0.002, 0.465±0.002, 0.331 ±0.004, 0.236± 0.003, 0.105±0.002 respectively）, and the differences had statistical significance （P〈0.01）; the expression of TGF-β, mRNA in rabbits＇ corneal flbroblasts could not only be inhibited by different OM concentrations, but its inhibition effect also was enhancing with the concentration increasing; α-SMA mRNA expression was not found in fibroblasts of normal rabbits＇ cornea. Conclusion： OM has inhibition effects on the proliferation of rabbits＇ corneal fibroblasts which cultured in vitro and the expression of TGF-β, mRNA, and this inhibition is enhancing with the OM concentration increasing.