不同浓度川芎嗪对人翼状胬肉成纤维细胞的作用
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【摘要】目的 观察川芎嗪对体外培养的人翼状胬肉成纤维细胞的抑制作用。方法收集眼科临床手术切除的人翼状胬肉标本，消化分离成纤维细胞，进行体外细胞培养。分别在pH为7.40和pH为6.00的条件进行，以无血清RPMI 1640培养液配制终浓度为4.0mg/ml、2.0mg/ ml、1.0mg/ ml、0.5mg/ ml和0.25mg/ ml的川芎嗪药液，0.02%的丝裂霉素C(MMC)作为阳性对照组，pH为7.40和/或pH为6.00的RPMI 1640培养液作正常对照组，对体外培养的胬肉成纤维细胞进行作用。通过光镜观察对细胞进行计数；四甲基偶氮唑盐(MTT)技术检测显示细胞存活率，间接免疫荧光技术检测细胞波形蛋白(Vimentin)的表达。结果 细胞计数和MTT法检测结果显示，在pH为7.40的条件下，与正常1640培养液组比较，4.0mg/ml的川芎嗪对体外培养的人翼状胬肉成纤维细胞均有明显的抑制作用(P<0.001)。在pH为6.00的条件下，与正常1640培养液组比较，不同浓度的川芎嗪和pH为6.00的1640培养液对人翼状胬肉成纤维细胞均有明显的抑制作用，与pH为6.00的1640培养液组比较，4.0mg/ml的川芎嗪对人翼状胬肉成纤维细胞的抑制作用明显强于其他浓度的川芎嗪，0.5mg/ml和0.25mg/ml的川芎嗪可明显促进人翼状胬肉成纤维细胞的生长(P<0.001)。结论川芎嗪对人翼状胬肉成纤维细胞的生长具有双重作用，较高浓度的川芎嗪(4.0mg/ml)对人翼状胬肉成纤维细胞的生长有抑制作用。较低浓度的川芎嗪(0.5mg/ml、0.25mg/ml)在pH为6.00的条件下对人翼状胬肉成纤维细胞的生长有保护作用。
【关键词】川芎嗪翼状胬肉成纤维细胞作用
【Abstract】Objective To investigate the effect of tetramethylpyrazine in different concentration on pterygium fibroblasts in vitro. Methods Human pterygium samples were collected and then digested and separated and cultured in vitro. Cultured human pterygium fibroblasts of the 3rd passage were treated by different concentration of tetramethylpyrazine under the condition of pH7.40 and pH6.00, 0.02% mitomycin ( MMC ) and normal 1640 cultured solution . The tetramethylpyrazine was diluted in different final concentration 4.0mg/ml, 2.0mg/ml, 1.0mg/ml, 0.5mg/ml and 0.25mg/ml using RPMI 1640 culture solution without fetal calf serum under the condition of pH 7.40 or 6.00, respectively. The cells were observed and counted under light microscope. The biological activities of human pterygium fibroblasts were evaluated by methyl thiazoly tetrazolium (MTT) assay. The identification of human pterygium fibroblasts was detected by indirect-immunofluorescence method. ResultsBy cell counting and MTT assay, compared with normal control group under the condition of pH 7 ......
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