Effects of 4 Thiopurine Compounds on Nitric Oxide Production and Cell Viability of HIG-82 Synoviocytes and RAW 264.7 Macrophages
Dahlan-Daud C. K1, Zain Z. N2, Tham C. L.1, Yong Y. K.3 and Hakim M. N1,41Department of Biomedical Sciences, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
2Faculty of Medicine and Health Sciences, Universiti Sains Islam Malaysia, Kuala Lumpur, Malaysia.
3Department of Human Anatomy, Faculty of Medicine and Health Sciences, Universiti Putra Malaysia, Serdang, Malaysia.
4Laboratory of UPM-MAKNA Cancer Research (CANRES), Institute of Bioscience, Universiti Putra Malaysia, 43400 UPM Serdang, Selangor, Malaysia.
Corresponding Author E-mail: nazrulh@upm.edu.my
Abstract: The 6-Mercaptopurine (6-MP) is a purine sulphur derivative used in the treatment of children with leukaemia as an antitumor medication. Nonetheless, the anti-proliferation and anti-rheumatoid arthritic activities of the drug have not been previously described. Therefore, the current study objective was to determine the cytotoxicity effects of 6-MP, 6-mercaptopurine riboside (6-MPR), 6-thioguanine (6-TG) and 6-thioxanthine (6-TX) on murine macrophages (RAW 264.7) and rabbit synoviocytes (HIG-82) cell lines. An inflammatory stage of RAW 264.7 and HIG-82 cell lines stimulated with Escherichia coli lipopolysaccharide (LPS) and phorbol-12-myristate acetate (PMA), respectively were treated with the drugs at a serials concentration 3.125 – 100 µM. Subsequently, the MTT assay was used to determine the viability of cells. The inhibitory effects were measured based on nitric accumulation in the conditioned media. The results showed that all drugs tested did not show any cytotoxic effect on both cell lines at low and medium concentrations, which the HIG-82 cell viability was more than 80%. However, the drugs display anti-proliferation property on RAW 264.7 cells compared to control. Reduction on cell proliferation was found on all tested drugs. Among its, the diclofenac significantly reduced the proliferation on HIG-82 cell compared to other drug compounds. Inhibitory effects of compound on nitric oxide production PMA-stimulated HIG-82 had only a small inductive effect, but excellent inhibition was observed on RAW 264.7 cell. Meanwhile, 100 μM 6-thioguanine and diclofenac had cytotoxic effect to RAW 264.7 and HIG-82 cell respectively. All thiopurines enhanced the proliferation of HIG-82 and RAW 264.7 cells with no cytotoxic effects. This finding opens new avenues for treating RA and the anti-rheumatic activities during synovial inflammation stage of RA. The inhibitory effects of 6-MP and 6-MPR to inflammatory cells marker such as synovial fibroblast and macrophages by proliferating healthy synoviocytes more potenly than 6-TG and 6-TX. The current study has demonstrated the potential use of 6-MP and its associated thio compounds in the in-vitro approach of HIG-82 cell culture in curing rheumatoid arthritis disease.
Keywords: Cytotoxicity; HIG-82; Nitric oxide; RAW 264.7 Back to TOC