Journal of Ethnopharmacology 138 (2011) 668–675 Contents lists available at SciVerse ScienceDirect Journal of Ethnopharmacology jo ur nal homep age : www.elsevier.com/locate/jethpharm Transcriptional profiling of human skin fibroblast cell line Hs27 induced by herbal formula Astragali Radix and Rehmanniae Radix Qi Zhang a , Fan Wei a , Chi Chun Fong a,b , Wai Kin Yu a , Yao Chen a,b , Chi Man Koon c,d , Kit Man Lau c,d , Ping Chung Leung c,d , Clara Bik San Lau c,d , Kwok Pui Fung c,d,e , Mengsu Yang a,b,∗ a Department of Biology and Chemistry, City University of Hong Kong, Kowloon, Hong Kong b Shenzhen Key Laboratory of Biochip Research, City University of Hong Kong, Shenzhen 518057, PR China c Institute of Chinese Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong d State Key Laboratory of Phytochemistry & Plant Resources in West China, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong e School of Biomedical Sciences, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong a r t i c l e i n f o Article history: Received 25 March 2011 Received in revised form 17 August 2011 Accepted 25 August 2011 Available online 30 October 2011 Keywords: Chinese herbal formula Astragali Radix Rehmanniae Radix cDNA microarry Wound healing a b s t r a c t Ethnopharmacological relevance: The herbs Astragali Radix (AR) and Rehmanniae Radix (RR) have long been used in traditional Chinese Medicine and serve as the principal herbs in treating diabetic foot ulcer. Aim of the study: Chinese herbal formulus comprising Astragali Radix (AR) and Rehmanniae Radix (RR) have been shown to improve the healing of diabetic foot ulcer through enhancing the viability of primary fibroblasts in diabetic patients suffering insulin resistance. Our previous study demonstrated that the herbal formula NF3 comprising of AR and RR in the ratio of 2:1 was effective in promoting wound healing in diabetic rats, and in vitro data indicated that the wound healing effects of NF3 might be due to the regulation and coordination of inflammation, angiogenesis and tissue regeneration. However, the under- lying molecular mechanism has not been well investigated. In this study, we investigated the cellular and molecular effects of the herbal formula NF3 on human skin fibroblast cells. Materials and methods: Human skin fibroblast cells Hs27 were treated with NF3 ranging from 0 to 8 mg/ml for 24 h, and the cells without NF3 treatment were used as control. Cell proliferation assay and cell cycle analysis were performed. Transcriptional profiles of Hs27 cells upon NF3 treatment were acquired by using a human cDNA microarray containing 10,000 genes, and the signaling pathways differentially regulated by NF3 were identified and analyzed. Results: NF3 promoted Hs27 cell proliferation and cell cycle progression. Microarray analysis revealed that 116 genes were differentially expressed upon NF3 treatment. Functional analysis of the genes indicated that NF3 mainly activated Wnt and angiogenesis related pathways, which are directly related to cell proliferation, angiogenesis, extracellular matrix (ECM) formation and inflammation during the process of wound healing. Conclusion: This study provides insight into the molecular mechanism of how the herbal formula Astragali Radix and Rehmanniae Radix may serve as potential therapeutics for wound healing. © 2011 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Chinese herbal formulus containing Astragali Radix (AR) and Rehmanniae Radix (RR) were found to promote the formation of Abbreviations: GLUT4, glucose transporter type 4; PDGFA, platelet-derived growth factor subunit A; BMP6, bone morphogenetic protein 6; COL1A1, collagen, type I, alpha 1; FZD1, frizzled-1; VEGF, vascular endothelial growth factor; Cyclin D1, G1/S-specific cyclin-D1; PDGF, platelet-derived growth factor; FGF, fibroblast growth factor. ∗ Corresponding author at: Department of Biology and Chemistry, City University of Hong Kong, Hong Kong. Tel.: +852 34427797; fax: +852 34420552. E-mail address: bhmyang@cityu.edu.hk (M. Yang). granulation tissue at the diabetic foot ulcer bed (Wong et al., 2001; Leung et al., 2008), and to stimulate the viability of primary fibrob- lasts in patients with insulin resistance (Lau et al., 2007, 2008, 2009). In particular, the innovative formula used in these studies, NF3 (AR:RR = 2:1, w/w), has been demonstrate to exhibit wound healing effects on the diabetic foot ulcer animal model (Tam et al., 2011). The results implied that NF3 was effective in promoting wound healing. However, the molecular mechanisms involved are not very well understood. Wound healing is a complex process involving several physio- logical events. Once a wound occurs, a number of different types of cells are recruited to participate in the healing process. Fibroblasts are predominant cells in the proliferation phase of wound healing, 0378-8741/$ – see front matter © 2011 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2011.08.080