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-PCR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(P<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。
[Objective]To observe the effects of pine pollen on the changes of p16INK4a and p21^WAF1 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, middledose pine pollen group（120mg/dl） and high-dose pine pollen group（240mg/dl）randomly. Regulate the phases of cell cycle by flow cytometry,and then the expression of p16INK4a, p21^WAF1 mRNA were determined by semi-quantitative RT-PCR. [Result]The amount of cells in G1 phase was depressed in middle-dose pine pollen group （72.73% ± 1.76%） and high-dose pine pollen group （64. 15±0.87%） than in old control group （81.33% ±3.43%）noticeablely （P〈0.05）. The expression of p16^INK4a mRNA 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,P〈0.05） ;The level of p21^WAF1 mRNA in middle-dose pine pollen group（0. 7692± 0. 1749） and high-dose pine pollen group （0. 4451±0. 0363） was significantly lower than in old control group（1. 1767±0. 2782,P 〈0.05）. [Conclusion]Pine pollen has the anti-aging effect on aging 2BS cells, this effect may be reacted via decreasing the ex- pression of p161NK4a and p21wg^WAF1 mRNA levels in aging cells, and then down regulating the amount of cells blocking in G1 phase of cell cycle.
出处
《浙江中医药大学学报》
CAS
2008年第1期89-92,共4页
Journal of Zhejiang Chinese Medical University
基金
国家自然科学基金资助项目(No:30271645)~~
