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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,研究骨科相关中药对雌激素受体(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pCMV-BD构建细胞杂交基因报告系统,研究骨科相关中药与雌激素受体的关系;MTT法检测中药对细胞活力的影响。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,部分中药可以显著提高细胞活性,并对雌激素受体具有激活作用。结论中药治疗骨质疏松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 hybridization gene reporter system containing estrogen receptor ligand binding domain and study the effect of orthopaedic related Chinese medicine 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 pCMV-BD to construct a cell hybridization gene reporting system to study the relationship between orthopedic traditional Chinese medicine and estrogen receptors; MTT assay was used to detect traditional Chinese medicines. Effect on cell viability. Results A mammalian cell hybrid gene reporter system containing the estrogen receptor ligand binding domain was successfully constructed. Some Chinese herbs can significantly increase cell viability and activate estrogen receptors. Conclusion Traditional Chinese medicine may play an important role in the treatment of osteoporosis through estrogen receptors. Cell hybridization gene reporter system can be used to study the mechanism of action of traditional Chinese medicine.