【關鍵詞】 肝細胞
［摘要］目的：觀察益氣、活血方藥對老年大鼠肝臟衰老相關基因表達的影響。方法：以自然衰老大鼠為衰老模型，採用RTPCR、Western blotting法研究益氣、活血方藥對老年大鼠肝臟p16、p21、增殖細胞核抗原（proliferating cell nuclear antigen, PCNA）、cyclin D1和cyclin E mRNA轉錄和蛋白表達的影響。結果：老年大鼠肝臟p16、p21、cyclin D1 mRNA轉錄和蛋白的表達增加；PCNA、cyclin E mRNA轉錄和蛋白的表達降低。與老年大鼠比較，益氣、活血方藥可下調老年大鼠肝臟cyclin D1 mRNA轉錄和蛋白表達，上調促進增殖基因cyclin E mRNA轉錄和蛋白表達，但對p16、p21和PCNA的調節作用不明顯。結論：益氣、活血方藥主要通過調控cyclin D1和cyclin E mRNA轉錄和蛋白表達而促進細胞增殖。
［關鍵詞］衰老大鼠; 肝細胞; p16; p21; 增殖細胞核抗原; cyclin D1; cyclin E; 益氣方; 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
隨著細胞及分子生物學的發展，已經發現一些與細胞衰老密切相關的基因及轉錄因子（反式作用因子）［1～3］。縱觀歷代中醫文獻，延緩衰老的方藥基本屬於補腎、健脾、益氣和活血範疇。近代研究顯示：上述四類方藥均有不同程度抗自由基、調節免疫功能、提高肝臟蛋白質含量、改善肝臟超微結構等延緩衰老的作用［4］。實驗研究還表明：補腎方藥能促進老年大鼠肝臟cHras、cmyc、cfos、cjun mRNA的表達，下調p53 mRNA的表達［5］。本實驗從細胞增殖調控角度，根據中醫理論，從益氣活血的`抗衰經典方中分別選擇幾味有代表性的中藥組成小複方，以自然衰老大鼠為衰老模型，應用RTPCR和Western blotting法研究益氣、活血方藥對老年大鼠肝臟衰老相關基因p16、p21、cyclin D1、增殖細胞核抗原(proliferating cell nuclear antigen, PCNA)和cyclin E表達的影響。