Min Yang1, Xin-Hua Qian, Dan-Hua Zhao, Su-Zhen Fu. 1. Department of Neonatology, Nanfang Hospital Affiliated Nanfang Medical University, 1838 Guang Zhou Da Dao North, Guangzhou 510515, Guangdong, PR China.
Abstract
ETHNOPHARMACOLOGICAL RELEVANCE: Astragalus polysaccharide (APS), obtained from Astragalus membranaceus, displays a range of activities in many systems, including the promotion of immune responses, anti-inflammation, and the protection of vessels. It possesses potent pharmacological activity on differentiation to the erythroid lineage. AIM OF THE STUDY: To investigate the effects of APS on the erythroid differentiation and the mechanism of action by microarray analysis in K562 cells. MATERIALS AND METHODS: Benzidine staining, semi-quantitative RT-PCR, Western blot and microarray methods were used to survey the effects of APS on inducing erythroid differentiation and the changes of gene expression profile in K562 cells. RESULTS: Of the 13.2% positive cells detected by benzidine staining, the induction was the highest with 200 microg/ml APS on 72h. Ggamma-mRNA expression and fetal hemoglobin synthesis were significantly up-regulated. Microarray analysis showed that 31 genes were up-regulated and 108 genes were down-regulated. These differential expression genes generally regulate protein binding, cellular metabolic process, the cell proliferation, and transcriptional activator activity. The gamma-globin gene was up-regulated, the genes related with erythroid differentiation such as LMO2, Runx1 and GTF2I were up-regulated, while Bklf, Eklf, EPHB4 and Sp1 were down-regulated. CONCLUSIONS: Our studies indicate that APS indicate potent activities on the erythroid differentiation by modulating genes of LMO2, Klf1, Klf3, Runx1, EphB4 and Sp1, increasing gamma-globin mRNA expression and fetal hemoglobin synthesis in K562 cells. Crown Copyright 2009. Published by Elsevier Ireland Ltd. All rights reserved.
ETHNOPHARMACOLOGICAL RELEVANCE: Astragalus polysaccharide (APS), obtained from Astragalus membranaceus, displays a range of activities in many systems, including the promotion of immune responses, anti-inflammation, and the protection of vessels. It possesses potent pharmacological activity on differentiation to the erythroid lineage. AIM OF THE STUDY: To investigate the effects of APS on the erythroid differentiation and the mechanism of action by microarray analysis in K562 cells. MATERIALS AND METHODS:Benzidine staining, semi-quantitative RT-PCR, Western blot and microarray methods were used to survey the effects of APS on inducing erythroid differentiation and the changes of gene expression profile in K562 cells. RESULTS: Of the 13.2% positive cells detected by benzidine staining, the induction was the highest with 200 microg/ml APS on 72h. Ggamma-mRNA expression and fetal hemoglobin synthesis were significantly up-regulated. Microarray analysis showed that 31 genes were up-regulated and 108 genes were down-regulated. These differential expression genes generally regulate protein binding, cellular metabolic process, the cell proliferation, and transcriptional activator activity. The gamma-globin gene was up-regulated, the genes related with erythroid differentiation such as LMO2, Runx1 and GTF2I were up-regulated, while Bklf, Eklf, EPHB4 and Sp1 were down-regulated. CONCLUSIONS: Our studies indicate that APS indicate potent activities on the erythroid differentiation by modulating genes of LMO2, Klf1, Klf3, Runx1, EphB4 and Sp1, increasing gamma-globin mRNA expression and fetal hemoglobin synthesis in K562 cells. Crown Copyright 2009. Published by Elsevier Ireland Ltd. All rights reserved.