补肾、健脾、益气、活血法对衰老细胞周期基因表达的调控作用

目的：比较补肾、健脾、益气、活血法对衰老细胞周期基因表达的调控作用。方法：采用上述4种不同的含药血清对衰老的人胚肺二倍体成纤维细胞2Bs细胞株进行处理，通过细胞寿命实验、流式细胞仪、RT—PCR、Westemblot方法研究不同中药对衰老细胞周期及其相关基因(P16^INK4、Cyclin D1及PCNA)的mRNA转录和蛋白表达的影响。结果：补肾、益气中药对衰老细胞周期有一定的促进作用，并可下调衰老细胞增殖抑制基因P16和周期蛋白Cyclin D1的mRNA／蛋白表达；上调细胞周期促进增殖基因PCNAmRNA／蛋白的表达。而健脾、活血中药对细胞周期及其相关基因和蛋白表达的作用不明显。结论：补肾、益气方药对细胞周期有促进作用，其中的作用可能是通过促进P16途径的基因表达来实现。

Comparative Study on the Regulatory Effects on Senescence Related Cell Cycle Gene Expression by TCM Prin- ciples of Tonifying Shen, Invigorating Pi Benefiting Qi, and Activating Blood Circulation

Objective: To compare the effects of TCM therapeutic principles of tonifying Shen (TS), benefiting Qi (BQ), invigorating Pi (IP) and activating blood circulation (ABC) herbs in regulating the gene expression in senescence related cell cycle. Methods: Drug sera containing TCM herbs of the above-mentioned principles were used to treat the aged human diploid fibroblast cell line 2BS. The effect of TCM on the senescence related cell cycle and its related gene expression (P16INK4 , Cyclin D1 and PCNA) were examined by means of cell prolifera-tive doublings, flow cytometry, RT-PCR and Western blot analysis. Results: TCM herbs of TS and BQ could improve the cell cycle, down-regulate the P16 and Cyclin Dl mRNA/protein expression, up-regulate PCNA mRNA/protein expression, while TCM herbs of IP and ABC showed insignificant effect on these indexes. Conclusion : TCM herbs of TS and BQ have effect in improving cell cycle, it may be achieved through promoting the P16 pathway of gene expression.