Study on high throughput screening method of identifying agonist for neuromedin U 2 receptor

ZHENG Xu-xu;OUYNG Ke-qing;GO Hong;XU Zhi-lung;HU Ying-he;CI Sho-xi;

Chinese Pharmaceutical Journal ›› 2004, Vol. 39 ›› Issue (03) : 185-188.

Chinese Pharmaceutical Journal ›› 2004, Vol. 39 ›› Issue (03) : 185-188.
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Study on high throughput screening method of identifying agonist for neuromedin U 2 receptor

  • ZHENG Xu-xu1;2;OUYANG Ke-qing1;GAO Hong1;XU Zhi-lung1;HU Ying-he1;CAI Shao-xi1;
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Abstract

OBJECTIVE To establish a high throughput screening method for finding agonist for NMU2R receptor METHODS The human NMU2R plasmid (hNMU2R/pCDNA3.1) and reporter gene plasmid (MRE/CRE/LUC) were cotransfected HEK293 to establish a cell line for agonist screening To identify and optimize the assay condition, the effects of some factors on the assay were examined by using forskolin and neuromedin U, such as incubation time, cell number in each well and the concentration of luciferase′s substrate The extracts of some traditional Chinese medicine were screened RESULTS A steady cell line and a reliable method for NMU2R agonist screening were established. The Z′-factor value was 0.75. The agonist incubation time was 6 to 8 hours. The cell number per well was 4×104to 8×104 CONCLUSION This technology was successfully applied to identify agonist for NMU2R receptor from the traditional Chinese medicines.

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NMU2R / gene / agonist / high throughput screening (HTS)

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ZHENG Xu-xu;OUYNG Ke-qing;GO Hong;XU Zhi-lung;HU Ying-he;CI Sho-xi;. Study on high throughput screening method of identifying agonist for neuromedin U 2 receptor[J]. Chinese Pharmaceutical Journal, 2004, 39(03): 185-188

References

［1］Shan LX,Qiao XD,Crona JH,et al. Identification of a novel neuromedin U receptor subtype expressed in the central nervous system［J］.J Bilo Chem,2000,275(50):39482. ［2］Howard AD,Wang RP,Pong SS,et al. Identification of receptors for neuromedin U and its role in feeding［J］.Nature,2000,406:70. ［3］郑旭煦，胡应和，欧阳克清,等. 神经介素U（neuromedin U）研究概况［J］. 生物化学与生物物理进展，2002，29（5）：765. ［4］Broach JR,Thorner J. Highthroughput screening for drug discovery［J］.Nature,1996,384(6604 Suppl):14. ［5］黄家学，胡娟娟，杜冠华. 化合物药物活性的高通量筛选［J］. 中国药理学通报,1999,16(5): 401. ［6］Major J. What is the future of high throughput screening?［J］.J Biomol Screen，1999,4(3):119. ［7］梁乾德，王升启. 药物高通量筛选分析技术［J］. 国外医学·药学分册，2002,29(1):22. ［8］Terstappen GC,Giacometti A,Ballini E,et al. Development of a functional reporter gene HTS assay for the identification of mGluR7 modulators［J］.J Biomol Screen, 2000,5(4):255. ［9］Jiang C,Chen G,Zeng X,et al. Generation of a bioactive neuropeptide in a cellfree system［J］.Anal Biochem,2003,316(1):34. ［10］Zhang JH,Chung TD,Oldenburg KR. A simple statistical parameter for use in evaluation and validation of high throughput screening assays［J］.J Biomol Screen,1999,4(2):67. ［11］Rosenberg D,Groussin L,Jullian E,et al. Role of the PKAregulated transcription factor CREB in development and tumorigenesis of endocrine tissues［J］.Ann N Y Acad Sci,2002,968:65. ［12］Dendorfer U,Oettgen P,Libermann TA. Multiple regulatory elements in the interleukin6 gene mediate induction by prostaglandins,cyclic AMP,and lipopolys accharide［J］.Mol Cell Biol,1994,14(7):4443.

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