益气活血方药对老年大鼠肝脏衰老相关基因表达的影响

【关键词】 肝细胞

［摘要］目的：观察益气、活血方药对老年大鼠肝脏衰老相关基因表达的影响。方法：以自然衰老大鼠为衰老模型，采用RTPCR、Western blotting法研究益气、活血方药对老年大鼠肝脏p16、p21、增殖细胞核抗原（proliferating cell nuclear antigen, PCNA）、cyclin D1和cyclin E mRNA转录和蛋白表达的影响。结果：老年大鼠肝脏p16、p21、cyclin D1 mRNA转录和蛋白的表达增加；PCNA、cyclin E mRNA转录和蛋白的表达降低。与老年大鼠比较，益气、活血方药可下调老年大鼠肝脏cyclin D1 mRNA转录和蛋白表达，上调促进增殖基因cyclin E mRNA转录和蛋白表达，但对p16、p21和PCNA的调节作用不明显。结论：益气、活血方药主要通过调控cyclin D1和cyclin E mRNA转录和蛋白表达而促进细胞增殖。

　　［关键词］衰老大鼠; 肝细胞; p16; p21; 增殖细胞核抗原; cyclin D1; cyclin E; 益气方; 活血方

　　Effects of recipes for replenishing qi and activating blood on senescence related gene expressions in the liver of aging rats

　　ABSTRACT Objective: To observe the effects of recipes for replenishing qi and activating blood on p16, p21, proliferating cell nuclear antigen (PCNA), cyclin D1 and cyclin E gene expressions in the liver of aging rats. Methods: A recipe for replenishing qi and a recipe for activating blood were administered to aging rats respectively, and the effects of the above recipes on the expressions of senescence related genes (p16, p21, PCNA, cyclin D1 and cyclin E) were examined by RTPCR and Western blotting methods. Results: The expressions of p16, p21 and cyclin D1 mRNAs and proteins in the liver of the untreated aging rats were upregulated, while the expressions of PCNA and cyclin E mRNAs and proteins decreased. As compared with the untreated aging rats, both recipes could downregulate the expressions of cyclin D1 mRNA and protein and upregulate the expressions of cyclin E mRNA and protein, but had no obvious effects on the expressions of mRNAs and proteins of p16, p21 and PCNA. Conclusion: Recipes for replenishing qi and activating blood can improve the liver cell proliferation of aging rats via downregulating the expressions of cyclin D1 mRNA and protein and upregulating the expressions of cyclin E mRNA and protein.

　　KEY WORDS aging rats; hepatocytes; p16; p21; proliferating cell nuclear antigen; cyclin D1; cyclin E; qibenefiting recipe; bloodactivating recipe

　　随着细胞及分子生物学的发展，已经发现一些与细胞衰老密切相关的基因及转录因子（反式作用因子）［1～3］。纵观历代中医文献，延缓衰老的方药基本属于补肾、健脾、益气和活血范畴。近代研究显示：上述四类方药均有不同程度抗自由基、调节免疫功能、提高肝脏蛋白质含量、改善肝脏超微结构等延缓衰老的作用［4］。实验研究还表明：补肾方药能促进老年大鼠肝脏cHras、cmyc、cfos、cjun mRNA的表达，下调p53 mRNA的表达［5］。本实验从细胞增殖调控角度，根据中医理论，从益气活血的`抗衰经典方中分别选择几味有代表性的中药组成小复方，以自然衰老大鼠为衰老模型，应用RTPCR和Western blotting法研究益气、活血方药对老年大鼠肝脏衰老相关基因p16、p21、cyclin D1、增殖细胞核抗原(proliferating cell nuclear antigen, PCNA)和cyclin E表达的影响。