Journal of Ethnopharmacology 118 (2008) 65–70 Contents lists available at ScienceDirect Journal of Ethnopharmacology journal homepage: www.elsevier.com/locate/jethpharm Inhibition of pro-inflammatory mediators and tumor cell proliferation by Anisomeles indica extracts Shih-Chuan Hsieh a , Shih-Hua Fang b , Yerra Koteswara Rao a , Yew-Min Tzeng a,∗ a Institute of Biotechnology, Chaoyang University of Technology, 168 Gifeng East Road, Wufeng 413, Taiwan, ROC b Department of Microbiology, School of Medicine, China Medical University, Taichung 400, Taiwan, ROC article info Article history: Received 20 March 2007 Received in revised form 24 January 2008 Accepted 8 March 2008 Available online 14 March 2008 Keywords: Anisomeles indica Anti-inflammatory Crude extracts Tumor cells proliferation inhibitory activities abstract Anisomeles indica (L.) Kuntze (Labiatae), is a traditional anti-inflammatory herb used in Taiwan. The aque- ous and methanolic extracts of whole plants, leaves, flowers and stems; and chloroform and n-butanol fractions of methanol extract, from A. indica were investigated for their anti-inflammatory activity on murine peritoneal macrophages. In addition, the tumor cells proliferation inhibition activities of these extracts were also evaluated against a panel of tumor cell lines such as Colon 205, PC 3, HepG2 and MCF 7. Treatment with A. indica extracts did not reduce cell viability at any dose used. However, all the extracts significantly inhibited the enhanced production of NO radicals, and pro-inflammatory cytokines (TNF-, and IL-12) induced by LPS/IFN- in a dose-dependent manner. Furthermore, methanolic extracts of leaves and flowers significantly and dose-dependently arrest mitogen-stimulated spleen cells in G0/G1 stage, in addition to their cell proliferation inhibition against Colon 205, MCF 7 and PC 3 by 94, 82; 98, 71; 82, 98%, respectively, at 200 g/mL concentration. This is the first report on A. indica extracts for their growth inhibitory activities, against inflammatory mediator production, and human tumor cell lines, colon, prostate, hepatoma and breast cells proliferation. © 2008 Elsevier Ireland Ltd. All rights reserved. 1. Introduction Macrophages play an important role in inflammatory disease through the release of factors such as free radical nitric oxide (NO), prostaglandin mediators, and cytokines involved in the immune response (Adams and Hamilton, 1984). Production of these macrophage mediators has been determined in many inflamma- tory tissues, following exposure to immune stimulants including bacterial endotoxin lipopolysaccharide (LPS) and interferon (IFN)- . Excessive production of the inflammatory mediators is involved in many diseases, such as rheumatoid arthritis (RA), chronic hep- atitis, and pulmonary fibrosis (Heinzel, 1990). Thus inhibition of inflammatory mediators’ production in response to inflammatory stimuli might be a useful therapeutic strategy in inflammatory dis- eases (Nam, 2006; Wang et al., 2007). On the other hand, cancer Abbreviations: Con A, concanavalin A; IFN, interferon; IL, interleukin; LPS, lipopolysaccharide; MB, n-butanol fraction of methanolic extract of whole plants; MC, chloroform fraction of methanolic extract of whole plants; MF, methanolic extract of flowers; ML, methanolic extract of leaves; MS, methanolic extract of stems; MWP, methanolic extract of whole plants; NO, nitric oxide; PEM, peritoneal excluded macrophages; RA, rheumatoid arthritis; TNF-, tumor necrosis factor-; WF, aque- ous extracts of flowers; WL, aqueous extracts of leaves; WS, aqueous extracts of stems; WWP, aqueous extracts of whole plants. ∗ Corresponding author. Tel.: +886 4 23323000x3003; fax: +886 4 23304896. E-mail address: ymtzeng@cyut.edu.tw (Y.-M. Tzeng). incidence and morbidity are steadily increasing in several parts of the world even though there is a decline in certain cancers. It was reported that natural anti-inflammatory agents may useful for the inhibition of certain tumor cells proliferation (Dufour et al., 2007). There is an increasing interest in herbal medications especially for chronic inflammatory diseases, in addition to their scaveng- ing free radicals and limit the initiation and progression of tumors (Dunn and Wilkinson, 2005; Nam, 2006). Anisomeles indica O. Kuntze (Labiate) is a perennial erect herb, 70–150cm tall, covered with septate hairs that the decoction of the stems is commonly used as tea and juice in Taiwan for the treatment of inflamma- tory diseases (Local name: Yu-Chen-Tsao) (Huang et al., 2003). It is used in folk medicine in the treatment of diverse conditions such as inflammatory skin diseases, liver protection, intestinal infections, abdominal pain and immune system deficiencies (Huang et al., 2003). Macrocyclic diterpenoids that showed the growth inhibit- ing activities against cultured KB cells were isolated from the whole plants of A. indica (Arisawa et al., 1986a,b). Acylated flavone gluco- sides, 6-methoxy flavones and HIV-inhibitory diterpenoid have also been extracted from aerial parts, stems and roots, respectively of A. indica (Ansari and Dobhal, 1982; Rao et al., 1983a,b, 1985; Dobhal et al., 1988; Shahidul et al., 2000). A decoction from the pre-flowering stage leaves and stems shown to has anti-histamine, free radical scavenging, membrane stabilizing, and cyclooxygenase-I inhibitory activities. In addition, the above aqueous extract also shown to has 0378-8741/$ – see front matter © 2008 Elsevier Ireland Ltd. All rights reserved. doi:10.1016/j.jep.2008.03.003