Synergistic modulation of proinflammatory mediators and cytokines in lipopolysaccharide-activated RAW 264.7 macrophages: The therapeutic potential of Elephantopus scaber and Sauropus androgynus ethanol extract.

Background and Aim: Elephantopus scaber (ES) and Sauropus androgynus (SA) have broad biological effects and have long been used in traditional medicine. However, the anti-inflammatory properties of the combination of ES and SA have not yet been fully explored. This study aimed to investigate the anti-inflammatory activities of the combination of ES and SA ethanol extract on lipopolysaccharide (LPS)-activated RAW 264.7 macrophage cell lines by inhibiting proinflammatory mediators and cytokines. Materials and Methods: Nitric oxide (NO) production in RAW 264.7 cells was assessed using the Griess protocol. The effects of the combination of ES and SA ethanol extract on RAW 264.7 cell viability were determined using WST-1 (4-[3-(4-Iodophenyl)-2-(4-nitro-phenyl)-2H-5-tetrazolio]-1,3-benzene sulfonate) assay. The levels of proinflammatory cytokines, including interferon-gamma (IFN-γ), tumor necrosis factor-alpha (TNF-α), and interleukin-1 beta (IL-ß), as well as the production of inducible nitric oxide synthase (iNOS), were assessed using flow cytometry. Results: This study demonstrated that ES and SA have excellent NO, iNOS, and proinflammatory inhibitory activities on LPS-induced RAW 264.7 macrophages. The formula ratio of 2ES:1SA showed the best NO inhibitory activity without any cytotoxicity, whereas the higher dose of SA (1ES:2SA) showed the best suppression of iNOS and proinflammatory cytokines IL-1ß, IFN-γ, and TNF-α. Conclusion: The combination of ES and SA ethanol extract could be an alternative agent for reducing excessive inflammation in inflammatory diseases.

Hydrophobic constituents of Polygonum multiflorum roots promote renal erythropoietin expression in healthy mice.

The roots of Polygonum multiflorum Thunberg (Polygonaceae) are used as a crude drug Kashu that is considered to improve blood deficiency based on a Kampo concept. Kashu has been included in Kampo formulas, such as Tokiinshi, which is used to treat eczema and dermatitis with itchiness by inhibiting inflammation and facilitating blood circulation in the skin. However, the effects of P. multiflorum roots on erythropoiesis are unclear. Previously, we isolated six phenolic constituents from an ethyl acetate (EtOAc)-soluble fraction of P. multiflorum root extract and identified them as (E)-2,3,5,4'-tetrahydroxystilbene-2-O-ß-D-glucopyranoside [(E)-THSG], emodin, emodin-8-O-ß-D-glucopyranoside, physcion, physcion-8-O-ß-D-glucopyranoside, and catechin. To examine whether P. multiflorum roots facilitate erythropoiesis, the EtOAc-soluble fraction was orally administered to healthy ICR mice. When compared with mice fed a standard diet alone (Controls), the mice fed a diet including the EtOAc-soluble fraction exhibited significantly higher serum erythropoietin (Epo) levels. The renal Epo mRNA levels in EtOAc-soluble fraction-administered mice were significantly higher than those in the control mice. Then, we administered roxadustat, which is a drug to treat the patient suffering with renal anemia by specifically inhibiting hypoxia-inducible factor prolyl hydroxylases. Roxadustat slightly increased renal Epo mRNA levels in healthy mice. Administration of (E)-THSG, a major constituent, significantly increased serum Epo levels. It is likely that (E)-THSG may facilitate the process to convert inactive renal Epo-producing cells to active Epo-producing cells. Collectively, it is implied that (E)-THSG in the EtOAc-soluble fraction of P. multiflorum roots may primarily improve blood deficiency of Kampo concept by promoting erythropoiesis.

Bitter melon fruit extract enhances intracellular ATP production and insulin secretion from rat pancreatic ß-cells.

Bitter melon (Momordica charantia L.) has been shown to have various health-promoting activities, including antidiabetic and hypoglycaemic effects. Improvement in insulin sensitivity and increase in glucose utilisation in peripheral tissues have been reported, but the effect on insulin secretion from pancreatic ß-cells remains unclear. In this study, we investigated the effect of bitter melon fruit on insulin secretion from ß-cells and the underlying mechanism. The green fruit of bitter melon was freeze-dried and extracted with methanol. The bitter melon fruit extract (BMFE) was fractionated using ethyl acetate (fraction A), n-butanol (fraction B) and water (fraction C). Insulin secretory capacity from INS-1 rat insulinoma cell line and rat pancreatic islets, as well as glucose tolerance in rats by oral glucose tolerance test (OGTT), was measured using BMFE and fractions. ATP production in ß-cells was also examined. BMFE augmented insulin secretion from INS-1 cells in a dose-dependent manner. The significant augmentation of insulin secretion was independent of the glucose dose. Fraction A (i.e. hydrophobic fraction), but not fractions B and C, augmented insulin secretion significantly at the same level as that by BMFE. This finding was also observed in pancreatic islets. In OGTT, BMFE and fraction A decreased blood glucose levels and increased serum insulin levels after glucose loading. The decrease in blood glucose levels was also observed in streptozotocin-induced diabetic rats. In addition, BMFE and fraction A increased the ATP content in ß-cells. We concluded that hydrophobic components of BMFE increase ATP production and augment insulin secretion from ß-cells, consequently decreasing blood glucose levels.

The role of VipAlbumin(®) as an immunostimulatory agent for controlling homeostasis and proliferation of lymphoid cells.

VipAlbumin(®) is a supplement from snakehead fish (Ophiocephalus striatus) which has high content of albumin that is very important to develop new cells. The aims of this study were to know the effect of VipAlbumin(®) to cell proliferation, expression level of CD4(+)CD62L(+) T cell, regulatory T cell, and B220+ cell, and immunocompetent cell cycle. Cell isolated from spleen of pathogen free mice were cultured in RPMI 1640 with 10% FBS, 1% Pen/Strep 10×, 2-Mercaptoetanol, anti-CD3 and LPS. The concentrations of VipAlbumin (®) used were 0 µg/ml; 0.33 µg/ml; 33.3 µg/ml; and 3333.3 µg/ml. The cell was incubated in CO2 5% incubator 37°C for 3 days for cell cycle and 5 days for proliferation analysis and cell expression. FACS analysis was done to know cell proliferation profile, status of cell, and cell cycle. Concentration 33.3 µg/ml and 3333.3 µg/ml significantly can increase cell proliferation and induce cell enter G2/M phase (p < 0.05) compared to control. VipAlbumin can significantly increase the relative number of CD4(+)CD62L(+) T cell, regulatory T cell, and B220+ cell (p < 0.05) compared to control. This study gives scientific evidence that VipAlbumin can be used as an immunostimulant which accelerates immunocompetent cells growth.