汉防己甲素抑制翼状胬肉成纤维细胞增殖的研究

Inhibitory effects of tetrandrine on pterygium fibroblasts

目的:观察汉防己甲素对体外培养的翼状胬肉成纤维细胞增殖抑制作用并探讨其机制,寻找治疗翼状胬肉和预防其复发的有效药物。方法:(1)倒置相差显微镜下观察不同浓度的Tet作用下,体外培养的翼状胬肉成纤维细胞形态的变化,并与丝裂霉素(Mitomycin,MMC)进行比较。(2)MTT比色法测定不同浓度的Tet作用48h对体外培养的翼状胬肉成纤维细胞增殖的抑制作用,并与MMC进行比较。(3)流式细胞仪检测不同浓度的Tet作用48h,体外培养的翼状胬肉成纤维细胞细胞周期的变化。结果:(1)Tet作用下,细胞胞体收缩,变圆,间隙增宽,药物浓度高时,细胞碎裂,浮起。0.1,0.2g/LMMC作用下大量细胞碎裂,溶解。(2)不同浓度的Tet作用48h对体外培养的翼状胬肉成纤维细胞的增殖均有明显的抑制作用(P<0.01);0.1g/L与0.2g/LMMC、4×10-5mol/LTet与0.2g/LMMC对翼状胬肉成纤维细胞活性的影响无显著性差异(P>0.05)。Tet的IC50接近10-5mol/L(为0.848×10-5mol/L)。(3)Tet作用下,翼状胬肉成纤维细胞周期中S期细胞增多,浓度越高,作用越明显。结论:汉防己甲素对翼状胬肉成纤维细胞的增殖具有明显的抑制作用,呈剂量依赖性,细胞分裂被阻止在S期(DNA合成期)。可望用于对翼状胬肉的治疗。

AIM： To investigate the inhibitory effects of tetrandrine （Tet） on the proliferation of cultured pterygium fibroblasts in vitro and its possible mechanism and to find a potentially effective agent for treatment of primary pterygium and prevention of its postoperative recurrence. METHODS： The cultured pterygium fibroblasts were exposed to different concentrations of Tat and MMC solution for 48 hours and investigated by MTT. The morphological changes of fibroblasts were observed through microscope. The changes of cell cycle were measured by flow cytometer （FCM）. RESULTS： Various concentrations of Tet solution have significant antiproliferative action on cultured pterygium fibroblasts （P〈0.01）. Fibroblasts exhibited morphological changes： shrinked to round or split into pieces when treated with high concentration of Tet solution. There was no statistical difference in absorbency between 0.1g/L MMC and 0.2g/L MMC tested groups, as well as 4×10^-5mol/L Tet and 0.2g/L MMC tested group （P 〉0.05）. The cell numbers distinctively increased in S phase when treated with high concentration of Tat solution. CONCLUSTON： Tet demonstrates an inhibitory effect in vitro on pterygium fibroblasts in culture and shows a dose-dependent manner. It may be used for treating and preventing postoperative recurrence of the common ocular disease.