Journal of Chinese Pharmaceutical Sciences ›› 2014, Vol. 23 ›› Issue (9): 617-625.DOI: 10.5246/jcps.2014.09.079

• Original articles • Previous Articles     Next Articles

Comparison of berberine and its five analogues on cell viability and COX-2 expression during glucose-oxygen deprivation and reperfusion in PC12 cells

Yunong Pang1, Jun Hu1, Yushuang Chai1, Hao Wu2, Yugang Wang1, Fan Lei1, Dongming Xing1, Lijun Du1*   

  1. 1. MOE Key Laboratory of Protein Sciences, Laboratory of Molecular Pharmacology and Pharmaceutical Sciences, School of Life Sciences and School of Medicine, Tsinghua University, Beijing 100084, China
    2. NGM Biopharmaceuticals, Inc., South San Francisco, California 94080, United States
  • Received:2014-04-14 Revised:2014-05-15 Online:2014-09-23 Published:2014-05-20
  • Contact: Tel./Fax: 86-10-62773630
  • Supported by:
    National Natural Science Foundation of China (Grant No. 81374006, 81073092 and 90713043) and the National S&T Major Special Project for New Drug R&D Program of China (Grant No. 2012ZX09103-201-041, 2012ZX09102-201-008 and 2011ZX09101-002-11).


Berberine, an isoquinoline alkaloid component of Rhizoma Coptidishas been demonstrated to be the key active ingredient involved in its protective effect against cerebral ischemia-reperfusion. However, the comparison among the analogues to the protective effect against oxygen and glucose deprivation/reoxygenation (OGD-R) was mediated by inhibition of cyclooxygenase-2 (COX-2) has never been reported. The aim of this study is to investigate the protective effect of berberine and its five analogues against OGD-R in PC12 cells, as well as to determine whether the protective effect was regulated through COX-2. An established in vitro OGD-R model of PC12 cells by oxygen glucose deprivation of 4 h and reperfusion of 24 h was used in our study. After cells were treated with berberine or its five analogues, we examined the cell viability assay by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay. Cells were also collected to determine the levels of mRNA and protein of COX-2 by real time PCR and Western blot. We found that berberine and its analogues improved the viability of PC12 cells against OGD-R. Whereas berberine and berberrubine presented stronger activity with the most effective dose of 0.31 μg/mL and the minimum effective doses of 0.02 and 0.04 μg/mL. Palmatine possessed potentially weaker protective effect. The mRNA level of COX-2 in cells treated with berberine, coptisine and epiberberine was decreased significantly. The protein level of COX-2 was significantly down-regulated in cells treated with berberine. Studies suggested the important role of methylenedioxy groups (R2 and R3) of berberine analogues in COX-2 inhibitory effect, and methylenedioxy groups (R2, R3, R9 and R10) in berberine analogues in binding affinity with COX-2. Substituted hydroxyl group at R9 did not affect the activity of berberine. In summary, our study illustrated the protective effects of berberine and its analogues in PC12 cells against OGD-R and to elucidate the structure-activity relationships. Docking analysis indicates that methylenedioxys at R2 and R3 is involved in the effect. More studies in other cells are needed to confirm our results.

Key words: Berberine, Analogues, COX-2, Structure-activity relationships

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