汉防己甲素对翼状胬肉成纤维细胞抑制作用的实验研究

Inhibitory effect of tetrandrine on pterygium fibroblasts in culture

目的观察汉防己甲素对体外培养的翼状胬肉成纤维细胞的抑制作用,寻找治疗翼状胬肉和预防其复发的有效药物。方法用MTT比色法测定不同浓度的汉防己甲素对体外培养的翼状胬肉成纤维细胞增生的抑制作用,并与丝裂霉素C(MMC)进行比较;用荧光定量PCR检测10-5mol/L的汉防己甲素对翼状胬肉成纤维细胞基质金属蛋白酶(MMP)MMP-3 mRNA的表达的影响。结果不同浓度的汉防己甲素对体外培养的翼状胬肉成纤维细胞的增生均有明显的抑制作用(P<0.01);10-5mol/L的汉防己甲素作用24 h其MMP-3 mRNA的表达量下降了27.12%。结论汉防己甲素对翼状胬肉成纤维细胞具有明显的抑制作用,可望用于对翼状胬肉的治疗。

Objective The anti-inflammation, anti-proliferation, anti-neovascularization, anti-tumor and anti-oxyzation effects of tetrandrine（Tet） have been determined in previous studies. Aims of this paper was to investigate the inhibitory effect of tetrandrine on cultured pterygium fibroblasts in vitro and find a potentially effective agent for treatment of primary and prevention of its postoperative recurrence. Methods Specimen of pterygium were collected during the operation and fibroblasts were cultured in free-serum medium added different concentrations of Tet （ 0. 25 × 10 ^- 5 , 0. 5 × 10^ - 5 , 1 × 10 ^- 5 , 2 × 10^ - 5 and 4 × 10^ - 5 mol/L） and mitomycin C （MMC,0.01% ,0.02% ） solution for 48 hours in experimental group,and pterygium fibroblasts in control group were cultured in only free-serum medium. Cultured pterygium fibroblasts were identified by immunochemistry. The effect of different concentration of Tet on cultured fibroblasts was evaluated using MTT. Expression of matrix metallopro-teinases 3 mRNA in fibroblasts was studied by Real-Time Fluorescent Quantitative PCR method. Results Cultured pterygium fibroblasts were specifically stained into brown. Cultured fibroblasts were significantly decreased in various concentrations of Tet groups and MMC groups in comparison with control group（ P 〈0. 01 ） , showing a strong antiproliferative action of Tet and MMC on cultured pterygium fibroblasts. The morphological changes of cultured fibroblasts included shrinking or splitting of cell in high concentrations of Tet group. There was no statistical difference in absorbency between 0. 01% MMC and 0.02% MMC groups or 4 × 10^ -5 mol/L Tet and 0.02% MMC groups （ P 〉 0.05 ）. Expression of MMP-3 mRNA on cultured pterygium fibroblasts in 10 -5 mol/L Tet group decreased by 27. 12%. Conclusion Tet has an inhibitory effect on cultured pterygium fibroblasts,and it may be useful for the treatment and prevention of recurrence of the common ocular disease.