【摘要】 目的探讨光敏剂血卟啉单甲醚(hematoporphyrinmonomethylether,HMME)介导的光动力学疗法(photodynamictherapy,PDT)对体外培养的人翼状胬肉成纤维细胞增殖的作用及其影响因素。方法用MTT比色法观察HMME对体外培养人初发、复发翼状胬肉成纤维细胞(以下简称为初发、复发细胞)增殖的影响(毒性作用);用MTT法测定不同的光敏剂孵育浓度(0、2.5、5.0、10、20和40μg/ml)、孵育时间(0、5、10、20、30和60min)和激光能量密度(0.62和2.835J/cm2)条件下行PDT对初发、复发细胞增殖的影响;用免疫组化法检测PDT对初发、复发细胞的增殖细胞核抗原(PCNA)表达的影响。结果初发、复发细胞分别在浓度≤20、40μg/ml的HMME中孵育60min对细胞无明显毒性。高剂量(2.835J/cm2)照光条件下,HMME孵育浓度分别≥2.5、10μg/ml的初发、复发细胞PDT实验组和对照组相比差异有显著意义(P<0.05),经40μg/ml的HMME孵育1h后用高剂量He-Ne激光照射初发、复发细胞增殖的抑制率分别为61.1%、57.6%;HMME孵育时间≥5min的PDT实验组和对照组相比差异有显著意义(P<0.01)。PDT对初发、复发细胞的PCNA表达具有明显的抑制作用。结论PDT对体外培养的初发、复发翼状胬肉成纤维细胞增殖有抑制作用。照光前光敏剂浓度越高、孵育时间越长,照光剂量越大,PDT效应越强。更多还原

【Abstract】 Objective To investigate the inhibition effect of photodynamic therapy with photosensitizer HMME on pterygium fibroblasts in vitro. Methods Cultured 3-5 passage primary and recurrent pterygium fibroblasts were treated with different concentration of HMME for different time, and the inhibitory rate of cells was measured by MTT colorimetric assay. The cells were divided into control group and experimental groups, after treated with different concentrations of photosensitizer(0,2.5,5.0,10,20,40μg/ml) under different doses of He-Ne laser(0.62,2.835J/cm~2)for different time(0,5,10,20,30,60min), all the cells were kept cultured for another 24 hours and the inhibitory rate and expression of proliferating cell nuclear antigen(PCNA) were examined by MTT colorimetric assay and immunohistochemical method, respectively. Results There were no significant effects on proliferation of primary and recurrent pterygium fibroblasts after they had been incubated for 60min in HMME less than 20,40μg/ml,respectively. He-Ne laser alone had no effect on pterygium fibroblasts. Under 2.835J/cm~2 He-Ne laser, there was statistic significance between PDT experimental groups and control group when the concentration of photosensitizer was not less than 2.5μg/ml and 10μg/ml in primary and recurrent pterygium fibroblasts respectively, and the inhibitory rate of primary pterygium fibroblasts was 61.1% in group of concentration photosensitizer and light dose with 40μg/ml and 2.835J/cm~2, respectively. The expression of PCNA in fibroblasts was decreased because of photodynamic therapy too. Conclusions Photodynamic therapy can inhibit the proliferation of pterygium fibroblasts in vitro. The inhibitory rate appears to be dependent on the effective time, concentration of photosensitizer and light dose in PDT.更多还原