Anti-Oxidation Mechanism of Andrographolide on HaCaT Cells Via Nrf2/ARE Pathway
HE Si-rong1,2, YU Huang-he1, YANG Zhen1, ZENG Rong1,2*
1. Department of Pharmacy, Hunan University of Chinese Medicine, Changsha 410208, China; 2. Hunan Key Laboratory of Druggability and Preparation Modification for Traditional Chinese Medicine, Hu��nan Province, Changsha 410208, China
Abstract��OBJECTIVE To investigate the anti-oxidant mechanism of andrographolide on HaCaT cells via Nrf2/ARE signal pathway. METHODS The effect of andrographolide on the viability of HaCaT cells and the effect of H2O2-induced cell viability were measured by CCK-8. HaCaT cells were pretreated with andrographolide of different concentration for 24 h. The protein and mRNA expression levels of Nrf2, HO-1, AKR1C1 and NQO1 in HaCaT cells were detected by Western blot and RT-qPCR, respectively. The expression of Nrf2 protein in the nucleus was analyzed by nuclear cytoplasmic separation and immunofluorescence. RESULTS Andrographolide had no significant effect on cell viability and dose-dependently decreased H2O2-induced cell death, the difference was statistically significant. Andrographolide significantly enhanced the expression of protein and mRNA of antioxidant enzymes Nrf2, HO-1, AKR1C1, NQO1, increased the distribution of Nrf2 in the nucleus, and up-regulated the expression of ARE. Besides, andrographolide upregulated the phosphorylation level of the upstream protein kinase AMPK�� (p-AMPK��). CONCLUSION Andrographolide could decrease H2O2-induced cell death, and its mechanism may be through the activation of Nrf2/ARE signaling pathway, thereby regulating the expression levels of HO-1, AKR1C1, and NQO1.
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2019, Vol. 54 
