Sekiguchi Yuusuke, Mano Hiroshi, Nakatani Sachie, Shimizu Jun, Kobata Kenji, Wada Masahiro
Department of Clinical Dietetics and Human Nutrition, Faculty of Pharmaceutical Sciences, Josai University, Sakado, Saitama, Japan.
BMC Res Notes. 2012 Apr 26;5:198. doi: 10.1186/1756-0500-5-198.
Salacia reticulata (SR) is a plant native to Sri Lanka. In ayurvedic medicine, SR bark preparations, taken orally, are considered effective in the treatment of rheumatism and diabetes. We investigated the ability of SR leaves (SRL) to inhibit in vitro the interleukin-1β (IL-1β)-activated proliferation of synoviocyte-like cells derived from rheumatoid arthritis model mice.
Inflammatory synovial tissues were harvested from type II collagen antibody-induced arthritic mice. From these tissues, a synoviocyte-like cell line was established and named MTS-C H7. To determine whether SRL can suppress cell proliferation and gene expression in MTS-C H7 cells, fractionation of the SRL hot-water extract was performed by high-performance liquid chromatography (HPLC), liquid-liquid extraction, sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE), and protease digestion.The 50% inhibitory concentration of the SRL hot-water extract against MTS-C H7 cells proliferation was ~850 μg/mL. Treatment with a low dose (25 μg dry matter per millilitre) of the extract inhibited IL-1β-induced cell proliferation and suppressed the expression of the matrix metalloproteinase (MMP) genes in MTS-C H7 cells. Various polyphenolic fractions obtained from HPLC and the fractions from liquid-liquid extraction did not affect cell proliferation. Only the residual water sample from liquid-liquid extraction significantly affected cell proliferation and the expression of MMP genes. The results of SDS-PAGE and protease digestion experiment showed that low molecular weight proteins present in SRL inhibited the IL-1β-activated cell proliferation.
We surmised that the residual water fraction of the SRL extract was involved in the inhibition of IL-1β-activated cell proliferation and regulation of mRNA expression in MTS-C H7 cells. In addition, we believe that the active ingredients in the extract are low molecular weight proteins.
网脉五层龙(SR)是一种原产于斯里兰卡的植物。在阿育吠陀医学中,口服SR树皮制剂被认为对治疗风湿和糖尿病有效。我们研究了SR叶(SRL)在体外抑制类风湿性关节炎模型小鼠来源的滑膜样细胞白细胞介素-1β(IL-1β)激活增殖的能力。
从II型胶原抗体诱导的关节炎小鼠中获取炎性滑膜组织。从这些组织中建立了一种滑膜样细胞系,并命名为MTS-C H7。为了确定SRL是否能抑制MTS-C H7细胞的增殖和基因表达,通过高效液相色谱(HPLC)、液-液萃取、十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白酶消化对SRL热水提取物进行了分级分离。SRL热水提取物对MTS-C H7细胞增殖的50%抑制浓度约为850μg/mL。用低剂量(每毫升25μg干物质)的提取物处理可抑制IL-1β诱导的细胞增殖,并抑制MTS-C H7细胞中基质金属蛋白酶(MMP)基因的表达。从HPLC获得的各种多酚级分和液-液萃取的级分均不影响细胞增殖。只有液-液萃取的残留水样显著影响细胞增殖和MMP基因的表达。SDS-PAGE和蛋白酶消化实验结果表明,SRL中存在的低分子量蛋白质抑制了IL-1β激活的细胞增殖。
我们推测SRL提取物的残留水级分参与了对IL-1β激活的细胞增殖的抑制以及MTS-C H7细胞中mRNA表达的调节。此外,我们认为提取物中的活性成分是低分子量蛋白质。