松花粉抗成纤维细胞复制性衰老的机制

目的 研究松花粉对衰老成纤维单细胞面积、β-半乳糖苷酶染色(SA-β-gal)阳性率、p16INK4A和p21CIP-1表达以及细胞周期的影响.方法 以二倍体成纤维(2BS)细胞建立衰老细胞模型.生物体视学法分析单细胞面积变化;免疫组化法测SA-β-gal阳性率;流式细胞术分析细胞周期;RT-PCR法测定p16INK4A、p21CIP-1基因mRNA表达量.结果 56代细胞出现典型的衰老细胞形态改变,同时SA-β-gal染色阳性率与G1期细胞比例均增高.经松花粉处理后,衰老细胞的单细胞面积、SA-β-gal染色阳性率与G1期细胞比例均显著减少(P＜0.05);p16INK4A与p21CIP-1mRNA的表达量均较衰老模型组明显下调(P＜0.05). 结论松花粉具有改善细胞复制性衰老的作用,其分子机制可能与下调p16INK4A及p21CIP-1基因mRNA的表达从而改善衰老细胞G1期阻滞有关.

Study on mechanism of pine pollen resisting cell replicative senescence in fibroblast

Objective To observe the effect of pine pollen on the surface area of single cell, the positive rate of senescence-associated (SA)-β-galactosidase stain, the expressions of p16INK4A and p21CIP-1 and the phase of cell cycle in 2BS cells. Methods The model of aging cell were made by 2BS cells. The surface area of single cell in each groups were analyzed by the method of biological stereology, the positive rate of SA-β-galactosidase stain was determined by immunohistochemistry,the phase of cell cycle was measured by flow cytometry, and the expressions of p16INK4A,p21CIP-1mRNA were detected by RT-PCR. Results The 56PD cell showed the typical shape changes of aging, the positive rate of SA-β-gal and the rate of the phase of G1 were both greatly increased (P＜0.05). After the treatment of pine pollen, the surface area of single cell, positive rate of SA-β-gal and the rate of the phase of G1 were significantly decreased (P＜0.05), the expressions of p16INK4A and p21CIP-1mRNA were both significantly down-regulated (P＜0.05). Conclusions Pine pollen has the anti-aging effect on aging 2BS cells, and its probable molecular mechanism may be related with down-regulating the expressions of p16INK4A and p21CIP-1mRNA and then improving the G1 phase blocking.