松花粉对人胚肺二倍体成纤维细胞p16及p21基因表达的影响_药学论文.docVIP

松花粉对人胚肺二倍体成纤维细胞p16及p21基因表达的影响_药学论文 松花粉对人胚肺二倍体成纤维细胞p16及p21基因表达的影响_药学论文 【摘要】 ［目的］研究松花粉对衰老细胞(2BS)内细胞周期蛋白激酶抑制剂p16INK4a及p21WAF1基因mRNA表达的影响。［方法］　2BS细胞从28代开始培养，随机分成年轻对照组（30代）、衰老模型组（56代）、松花粉中剂量（120mg/dl）组（56代）、松花粉高剂量(240mg/dl)组（56代）。流式细胞术分析细胞周期，检测G1期比例；半定量RT法测定p16INK4a、p21WAF1mRNA的表达。［结果］衰老模型组细胞出现典型的细胞体积增大，形态不规则；松花粉处理组细胞，体积回缩，形态趋于规则。G1期细胞比例松花粉中剂量组(72.73%±1.76%)和高剂量组(64.15%±0.87%)较衰老模型组(81.33%±3.43%)显著下降(0.05)。细胞周期蛋白激酶抑制剂p16INK4amRNA的表达量在松花粉中剂量组(0.3483±0.0078)和高剂量组(0.3222±0.0055)较衰老模型组(0.3974±0.0078)表达灰阶明显下调。p21WAF1mRNA的表达量在松花粉中剂量组(0.7692±0.1749)和高剂量组(0.4451±0.0363)较衰老模型组(1.1767±0.2782)。［结论］松花粉具有改善细胞衰老的作用，其分子机制可能与下调衰老细胞内细胞周期蛋白激酶抑制剂p16INK4a及p21WAF11mRNA的表达从而改善衰老细胞G1期阻滞有关。 【关键词】 松花粉;细胞衰老;细胞周期;p16;p21 　　Abstract：［Objective］To observe the effects of pine pollen on the changes of p16INK4a and p21WAF1 in 2BS cells.［Method］The 2BS cells were cultured from 28 population doubling(PD) and divided equally into the young control group, old control group, middleses of cell cycle by flow cytometry，and then the expression of p16INK4a，p21WAF1 mRNA were determined by semi［Result］The amount of cells in G1 phase was depressed in middle group (64.15%±0.87%) than in old control group (81.33%±3.43%)noticeablely (0.05).The expression of p16INK4amRNA in middledose pine pollen group(0.3483±0.0078) and highdose pine pollen group (0.3222±0.0055) was significantly decreased than in old control group(0.3974±0.0078,0.05)；The level of p21WAF1mRNA in middle［Conclusion］Pine pollen has the antiging effect on aging 2BS cells, this effect may be reacted via decreasing the expression of p16INK4a and p21WAF1mRNA levels in aging cells, and then down regulating the amount of cells blocking in G1 phase of cell cycle. 　　Key words：pine pollen；cell aging；cell cycle；P16；P21 　　细胞复制性衰老是目前普遍接受的细胞衰老理论，其最显著的特征即细胞出现G1期阻滞。在细胞衰老过程中细胞周期蛋白依赖激酶抑制因子（CDKI）发生表达量的改变。其中p16INK4a及p21WAF1是细胞衰老诱导途径中的两个关键产物，被称为衰老相关基因［1］。其表达量与衰老的表型及衰老进程密切相关，作为检测细胞衰老的分子指标应用于衰老相关检测。松花粉是天然的营养宝库，已有的研究发现，松花粉对人胚肺二倍体成纤维细胞有促进增殖［2］，并提高端粒酶活性［3］的作用。本实验探讨松花粉对衰老2BS