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

doi:10.5246/jcps.2014.09.079

Abstract

Abstract:

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 R₃is involved in the effect. More studies in other cells are needed to confirm our results.

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

CLC Number:

R963

Supporting:

Yunong Pang, Jun Hu, Yushuang Chai, Hao Wu, Yugang Wang, Fan Lei, Dongming Xing, Lijun Du. Comparison of berberine and its five analogues on cell viability and COX-2 expression during glucose-oxygen deprivation and reperfusion in PC12 cells[J]. Journal of Chinese Pharmaceutical Sciences, 2014, 23(9): 617-625.

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