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目的:构建含有雌激素受体配体结合域的哺乳动物细胞杂交基因报告系统,研究通络生骨胶囊对雌激素受体(ER)活性的影响。方法:PCR扩增雌激素受体配体结合域,将配体结合域克隆到真核表达质粒pCMVBD构建细胞杂交基因报告系统,研究通络生骨胶囊与雌激素受体的关系;MTT法检测通络生骨胶囊对细胞活力的影响。结果:成功构建含有雌激素受体配体结合域的哺乳动物细胞杂交基因报告系统,通络生骨胶囊可以显著提高细胞活性,并对雌激素受体具有激活作用。结论:通络生骨胶囊可能通过雌激素受体发挥疗效,细胞杂交基因报告系统可以用来研究中药的作用机理。
OBJECTIVE: To construct a mammalian cell hybrid gene reporter system containing the estrogen receptor ligand binding domain and study the effect of Tongluo Shenggu capsule on estrogen receptor (ER) activity. METHODS: The estrogen receptor ligand binding domain was amplified by PCR, and the ligand binding domain was cloned into the eukaryotic expression plasmid pCMVBD to construct a cell hybridization gene reporter system to study the relationship between Tongluo Shenggu capsule and estrogen receptors; MTT assay Effects of Tongluo Shenggu Capsule on Cell Viability. RESULTS: A mammalian cell hybrid gene reporter system containing the estrogen receptor ligand binding domain was successfully constructed. Tongluo Shenggu Capsule can significantly increase cell viability and activate estrogen receptors. Conclusion: Tongluo Shenggu Capsule may exert its therapeutic effect through estrogen receptor. Cell hybridization gene report system can be used to study the mechanism of action of traditional Chinese medicine.