Establishment and Application of High-throughput Screening Model for Active Ingredients of Traditional Chinese Medicine Based on CXCR4 Promoter
10.3969/j.issn.1005-5304.2018.11.012
- VernacularTitle:基于CXCR4启动子的中药有效成分高通量筛选细胞模型构建和应用
- Author:
Qiang-Li WANG
1
;
Hai-Dong GUO
;
Yi WANG
;
Xin-Yu WU
;
Liang ZHAO
;
Ying WANG
Author Information
1. 上海中医药大学基础医学院
- Keywords:
myocardial infarction;
stem cell transplantation;
CXCR4/SDF-1 axis;
luciferase reporter gene;
cell model
- From:
Chinese Journal of Information on Traditional Chinese Medicine
2018;25(11):52-56
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish a cell model targeting the CXCR4 gene promoter to screen of drugs potentially promoting the stem cell homing, and to screen the monomers of traditional Chinese medicine. Methods The human CXCR4 gene promoter sequence (279 bp) was inserted into the luciferase reporter vector pGL4.17-Basic to construct the recombinant plasmid pGL4.17-CXCR4, which was co-transfected with the internal reference plasmid pRL-TK into 293 cells. After verifying transcriptional activity of CXCR4 promoter, single cell clones were cultured to obtain a stable transfectant cell line and used for screening of candidate Chinese herbal monomer (1 to 10). Finally, Western blot was used to verify the effect of the selected Chinese herbal monomer on the expression of CXCR4 in MSCs. Results The drug screening cell model was successfully established and the candidate monomer 6 was screened out because of the strongest effect on enhancing CXCR4 promoter activity. Western blot results verified that the candidate monomer 6 could promoted the expression of CXCR4 of MSCs. Conclusion The drug screening cell model established in this study was able to achieve high-throughput screening for potentially promoting the stem cell homing Chinese herbal monomer.
