藤茶活性成分二氢杨梅素对明胶酶的影响

Effect of active ingredient dihydromyricetin from Ampelopsis grossedentata on gelatinases

该文探讨藤茶活性成分二氢杨梅素（DMY）对明胶酶酶活性和酶蛋白表达量的影响。通过收集不同质量浓度DMY处理的人乳腺癌细胞的条件化培养液上清液,取等蛋白质含量各处理组样本液,进行相同条件的明胶酶谱实验,以此检测DMY对明胶酶蛋白表达量的影响;收集未经过DMY处理的细胞培养液上清液,等量体积上样进行明胶酶谱实验的SDS-PAGE电泳部分,移除凝胶中SDS后置于含有不同质量浓度DMY的孵育液中进行孵育,以此检测DMY对明胶酶活性的影响。结果表明,20-80μg/mL的DMY对明胶酶活性未见明显影响,但以剂量依赖方式抑制明胶酶蛋白的表达。这表明DMY抑制肿瘤转移的作用机制可能是通过抑制明胶酶酶蛋白的表达量,而不是通过抑制该酶活性来完成的。

The effect of dihydromyricetin（DMY） on gelatinase activities and gelatinase proteins expression was investigated. Human breast cancer cells were treated by different concentrations of DMY. The conditioned media were collected, and equal amount of proteins sample were analyzed by gelatin zymography, which helped to detect the effect of DMY on gelatinase proteins expression. Supernatant without DMY pretreatment were collected, and equal volume samples were loaded to conduct SDS-PAGE part of the gelatin enzyme spectrum experiment. After removal of SDS, the gels were incubated in incubation buffer containing different concentrations of DMY, by which to detect the effect of DMY on gelatinase activities.Result showed that 20-80 μg/ml DMY showed no obvious effect on gelatinases activities, but inhibited the gelatinases proteins expression in a dose-dependent manner. The DMY＇s anti-tumor mechanism could be by restraining the gelatinases proteins expression, rather than by inhibiting the gelatinase activities.