多项研究证实六味地黄丸具有确切的预防癌前上皮恶变的作用，但具体机制尚未明确。新近研究发现衰老成纤维细胞分泌的SASP能促进癌前上皮恶性转化，延缓细胞衰老可预防癌的发生。祖国医学认为肾虚是衰老之本，现有研究证实中医的“肾虚”与细胞衰老密切相关，项目组最新研究提示六味地黄丸可通过抑制P21表达延缓成纤维细胞衰老，据此提出假说：补肾可干预衰老成纤维细胞诱导的癌前上皮恶变。本项目选用滋阴补肾、延寿抗衰的经方六味地黄丸，从延缓成纤维细胞衰老、干预衰老细胞SASP的新视角，采用蛋白芯片、蛋白印迹等分析衰老通路关键信号分子P16、P53等指标的变化，揭示该方抑制衰老成纤维细胞分泌SASP的分子靶点；运用细胞共培养技术、构建裸鼠肿瘤模型，体内外实验相结合，检测其对衰老成纤维细胞介导的癌前上皮恶性转化的拮抗作用。本研究将为中医“治未病”理论在防癌领域的应用提供新思路，为中医药预防癌的临床推广提供实验依据。

Studies have shown the positive role of Liu Wei Di Huang pills in preventing precancerous epithelial malignant, but its concrete mechanism is not yet clear. Recent studies have found that SASP secreted by aging fibroblasts can promote epithelial malignant transformation and slow down cell aging, thus preventing cancer. According to Chinese Medicine, kidney deficiency is the root cause of aging, and the existing researches have also proved that kidney deficiency is closely related to cell aging. This reserch group have resently found out that Liu Wei Di Huang pills can delay fibroblasts senescence by inhibiting P21. Hereby the group members have proposed that nourishing kidney can intervene precancerous epithelial malignant transformation caused by aging fibroblasts. In this research, Liu Wei Di Huang pill, a classical prescription of Chinese medicine nourishing Yin and strengthening kidney,is used to slow down the cell aging process and intervene SASP from a new perspective. The group will adopt Protein chip technology and Western blotting to analyze the changes of some key signal molecules such as P16 and P53 in order to find out the molecular targets of SASP secreted by aging fibroblast when the prescribtion is used . It invites cell cultural techniques and creats nude mice heterotransplanted models;the group members conduct experiments both in vivo and in vitro to detect drug antagonism of aging fibroblast’s effects on precancerous epithelial malignant transformation.The reseach intends to provide new methods for “preventive treatment of disease” in the application of Chinese medicine to prevent cancer,and lay an experimental foundation for the clinical application of Chinese medicine to prevent cancer.

多项研究证实滋阴补肾、延寿抗衰的经方六味地黄丸具有确切的预防癌前上皮恶变的作用，但具体机制尚未明确。新近研究发现衰老成纤维细胞分泌的SASP能促进癌前上皮恶性转化，延缓细胞衰老可预防癌的发生。祖国医学认为肾虚是衰老之本，现有研究证实中医的“肾虚”与细胞衰老密切相关，据此提出假说：补肾可干预衰老成纤维细胞诱导的癌前上皮恶变。本研究采用血清药理学方法制备含药血清，用叔丁基过氧化氢诱导成纤维细胞衰老，采用β-半乳糖苷酶染色、荧光定量RT-PCR等方法证实六味地黄丸能预防成纤维细胞衰老，其机制可能与抗氧化应激有关；运用细胞因子蛋白芯片技术检测六味地黄丸含药血清对衰老成纤维细胞分泌SASP的影响，筛选出其作用靶点为IL-6，并用ELISA、荧光定量RT-PCR等进行验证；通过蛋白印迹、平板克隆形成、划痕实验检测六味血清处理后的衰老HS68细胞培养上清对癌前细胞（Hacat、L02）上皮-间质转化、克隆形成、迁移等的影响，证实其对衰老成纤维细胞介导的癌前上皮恶性转化具有拮抗作用。另外我们还进行了与衰老相关的结肠癌、乳腺癌诊断标记物的研究：检索数据库进行肠癌相关目的基因的筛选，筛选出潜在肠癌诊断标记物基因Adenylyl cyclase 9 (ADCY9)、乳腺癌诊断标记物基因ROCK。采用组织芯片免疫组化方法证实ADCY9表达增强与结肠癌侵袭行为和预后密切相关，可成为潜在的评估肠癌患者预后的指标。本研究将为中医“治未病”理论在防癌领域的应用提供新思路，为中医药预防癌的临床推广提供实验依据。