【關(guān)鍵詞】 肝細(xì)胞

［摘要］目的：觀察益氣、活血方藥對老年大鼠肝臟衰老相關(guān)基因表達(dá)的影響。方法：以自然衰老大鼠為衰老模型，采用RTPCR、Western blotting法研究益氣、活血方藥對老年大鼠肝臟p16、p21、增殖細(xì)胞核抗原（proliferating cell nuclear antigen, PCNA）、cyclin D1和cyclin E mRNA轉(zhuǎn)錄和蛋白表達(dá)的影響。結(jié)果：老年大鼠肝臟p16、p21、cyclin D1 mRNA轉(zhuǎn)錄和蛋白的表達(dá)增加；PCNA、cyclin E mRNA轉(zhuǎn)錄和蛋白的表達(dá)降低。與老年大鼠比較，益氣、活血方藥可下調(diào)老年大鼠肝臟cyclin D1 mRNA轉(zhuǎn)錄和蛋白表達(dá)，上調(diào)促進(jìn)增殖基因cyclin E mRNA轉(zhuǎn)錄和蛋白表達(dá)，但對p16、p21和PCNA的調(diào)節(jié)作用不明顯。結(jié)論：益氣、活血方藥主要通過調(diào)控cyclin D1和cyclin E mRNA轉(zhuǎn)錄和蛋白表達(dá)而促進(jìn)細(xì)胞增殖。

　　［關(guān)鍵詞］衰老大鼠; 肝細(xì)胞; p16; p21; 增殖細(xì)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(xì)胞及分子生物學(xué)的發(fā)展，已經(jīng)發(fā)現(xiàn)一些與細(xì)胞衰老密切相關(guān)的基因及轉(zhuǎn)錄因子（反式作用因子）［1～3］�？v觀歷代中醫(yī)文獻(xiàn)，延緩衰老的方藥基本屬于補(bǔ)腎、健脾、益氣和活血范疇。近代研究顯示：上述四類方藥均有不同程度抗自由基、調(diào)節(jié)免疫功能、提高肝臟蛋白質(zhì)含量、改善肝臟超微結(jié)構(gòu)等延緩衰老的作用［4］。實(shí)驗(yàn)研究還表明：補(bǔ)腎方藥能促進(jìn)老年大鼠肝臟cHras、cmyc、cfos、cjun mRNA的表達(dá)，下調(diào)p53 mRNA的表達(dá)［5］。本實(shí)驗(yàn)從細(xì)胞增殖調(diào)控角度，根據(jù)中醫(yī)理論，從益氣活血的`抗衰經(jīng)典方中分別選擇幾味有代表性的中藥組成小復(fù)方，以自然衰老大鼠為衰老模型，應(yīng)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(guān)基因p16、p21、cyclin D1、增殖細(xì)胞核抗原(proliferating cell nuclear antigen, PCNA)和cyclin E表達(dá)的影響。