Resveratrol (3, 5, 4′-trihydroxy-trans-stilbene), a phytoalexin found in grapes and other dietary and medicinal plants, has been shown to have anti-inflammatory, antioxidant and antitumor activities[1-7]. the growth of H22 tumor in Balb/c mice. The antitumor effect of resveratrol might be related to directly Rabbit Polyclonal to OR2AG1/2 inhibiting the growth of H22 cells and indirectly inhibiting its potential effect on nonspecific host immunomodulatory activity. INTRODUCTION Recently, considerable attention has been focused on identifying naturally occurring chemopreventive substances capable of inhibiting, retarding, or reversing the multi-stage carcinogenesis. Resveratrol (3, 5, 4′-trihydroxy-trans-stilbene), a phytoalexin found in grapes and other dietary and medicinal plants, has been shown to have anti-inflammatory, antioxidant CX-6258 hydrochloride hydrate and antitumor activities[1-7]. Many of these beneficial effects of resveratrol require participation of the cells of the immune system. However, the effect of resveratrol on the development of immuological responses remains unknown. In the present study, the antitumor activity and immunomodulatory actions of resveratrol, including Magainst H22 cells, serum IgG and the plaque forming cells and tumor necrosis factor (TNF-) content in Balb/C mice with experimentally implanted tumor of H22 were investigated. MATERIALS AND METHODS Materials Resveratrol, CX-6258 hydrochloride hydrate MTT, IPS and dimethylsulfoxide (DMSO) were purchased from SGMA Co. Mouse hepatocellular carcinoma cells H22, L929 and sheep red blood cell (SRBC) were kindly supplied by Cheng Wei (Center of Molecular Biology, First Affiliated Hospital, Xi’an Jiaotong University). Cells were subcultured in RPMI 1640 (Gibco) which was supplemented with 10% fetal bovine serum, penicillin (100 IU?mL-1) and streptomycin (100 mg?l-1). Stock solution of resveratrol was made in dimethylsulfoxide (DMSO) at a concentration of 10 g?l-1. Working dilutions were directly made in the tissue culture medium. [3H]TdR was purchased from Shanghai Nuclear Research Institute. IL test kit and LPS were purchased from Gibco Co. Balb/C mice, 2.5 month old, weighing 202 g, were purchased from the Animal Center, Xi’an Jiaotong University. Methods Effect of resveratrol on cytotoxicity of peritoneal macrophages (M) against H22 cells M was collected from the peritoneal cavity of Balb/c mice 3 d after ip 10% sheep red blood cells (SRBC, 1 mL/mouse). CX-6258 hydrochloride hydrate The cells were washed twice and resuspended in RPMI 1640 culture medium. H22 cells were cultured for 12 h, and 100 L M suspension and different concentrations of resveratrol were added to each well of 96-well plates at a ratio of effectors: target (E:T) cell 10:1 or 25:1. After cultured for 24 h, each well was added with [3H]TdR (9.3 kBq/well), and then was incubated for another 6 h. Cells were placed onto the glass fiber filter and [3H]TdR incorporation was determined by liquid scintillation. The cytotoxicity was calculated with the following formula: the cytotoxicity of M = (control-treatment)/control 100% (dpm). Anti-tumor activity of resveratrol and its effect on serum antibody IgG, plaque CX-6258 hydrochloride hydrate forming cells (PFC) in Balb/C mice with implanted tumor of H22 Mouse ascites (including 2 105 cells) were injected into the right axilla of 40 Balb/c mice. On the second day, 40 Balb/c mice were divided into 4 groups randomly, and then were injected with resveratrol at a dose of 500 mg?kg-1, 1000 mg?kg-1, 1500 mg?kg-1 and normal saline for 10 d. Mice were sensitized to ip SRBC (3 107 cells). After 4 d, the mice were bled to obtain serum for IgG investigation. At the same time, spleens were excised for PFC counting. IgG contents were determined by single immunodiffusion method. PFC was measured by modified Cunningham method. CX-6258 hydrochloride hydrate Effect of resveratrol on serum tumor necrosis factor alpha (TNF-) production induced by LPS in Balb/c mice Ascites cells of 2 105 were injected into the Balb/c mice. Resveratrol at a dose a 500 mg?kg-1, 1000 mg?kg-1 and 1500 mg?kg-1 was injected for 10 d, and BCG of 200 mg?kg-1 as a positive control agent was.