摘要
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：The establishment of Gal4DBD-ERα or ERβ-LBD reporter system to study the effect of Chinese medicine on ERs. Methods：Gene reporter system was generated by amplifying the ligand binding domain（LBD） with Pfu polymerase and subcloning the product into the eukaryotic expression vector pCMV-BD,This system was employed to study the relation between Chinese medicine and ERs. cell activity were detected by MTT. Result：Gene reporter system using ER LBD is established, Some Chinese medicine can activate ERs and improve cell activation. Conclusion： The mechanisms of Chinese medicine of treating osteoporosis may be related to its effect of activating ERs,Gene reporter system can be used to study the mechanisms of Chinese Traditional Medicine.
出处
《中国中医骨伤科杂志》
CAS
2006年第6期43-45,共3页
Chinese Journal of Traditional Medical Traumatology & Orthopedics
基金
国家自然科学基金(基金号:30400606)
广东省自然科学基金(基金号:04010036)
广东省科技厅(基金号:粤科社字2004
139号)
广州中医药大学校创新基金(基金号:K004044)资助。
关键词
中药
基因报告系统
雌激素受体
HELA细胞
配体结合域
Chinese Medicine
Gene Reporter System
Estrogen Receptor
H eLa Cell
Ligand Binding Domain
