• Archives of Pharmacal Research
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• 제29권12호
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• pp.1147-1153
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• 2006

Mizoribine (MZR) has been shown to possess immunosuppressive activity that selectively inhibits the proliferation of lymphocytes by interfering with inosine monophosphate dehydrogenase. The efficacy of MZR is not only in patients who have had renal transplantation, but also in patients with rheumatoid arthritis (RA), lupus nephritis, and primary nephritic syndrome. Because the exact mechanism of its immunosuppressive action is not clear, the object of this study was to examine the ability of MZR to regulate the antigen presenting cells (APCs), dendritic cells (DCs). In this work, we tested whether MZR ($1{\sim}10\;{\mu}g/mL$) could inhibit the cross-presentation of DCs. DC2.4 cells ($H-2K^{b}$) or bone marrow-derived DCs (BM-DCs) generated from BM cells of C57BL/6 mouse ($H-2K^{b}$) were cultured in the presence of MZR with OVA-microspheres, and the amount of OVA peptide-class I MHC complexes was measured by a T cell hybridoma, B3Z, that recognizes OVA (257-264 : SIINFEKL)-$H-2K^{b}$ complex and expresses-galactosidase. MZR profoundly inhibited the expression of SIINFEKL-$H-2K^{b}$ complexes. This inhibitory activity of MZR appeared to affect the phagocytic activity of DCs. MZR also decreased IL-2 production when we examined the effects of MZR on $CD4^{+}$ T cells. These results provide an understanding of the mechanism of immunosuppressive activity of MZR on the inhibition of MHC-restricted antigen presentation and phagocytic activity in relation to their actions on APCs.

• 대한한의학회지
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• 제24권2호
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• pp.138-147
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• 2003

Objectives : In this study, we investigated the mechanism by which Chelidonium majus (CM) regulates nitric oxide (NO) production. Methods : Using mouse peritoneal macrophages, the mechanism by which CM regulates NO or tumor necrosis $factor-{\alpha}(TNF-{\alpha})$ production was examined. NO release was measured by the Griess method. $TNF-{\alpha}$ production was measured by the ELISA method. The protein extracts were prepared and samples were analyzed for the inducible NOS(iNOS) expression and nuclear factor kappa $B(NF-{\kappa}B)$ activation by Western blotting. Results : When CM was used in combination with recombinant $interferon-{\gamma}{\;}(rIFN-{\gamma})$, there was a marked cooperative induction of NO production. CM had an effect on NO production by itself. The expression of the iNOS gene was increased in $rIFN-{\gamma}$ plus CM-stimulated peritoneal macrophages and almost completely inhibited by pre-treatment with pyrrolidine dithiocarbamate (PDTC), an inhibitor of $NF-{\kappa}B$. The $NF-{\kappa}B$ activation was increased in rIFN-{\gamma} plus CM-induced peritoneal macrophages. The increased production of NO from $rIFN-{\gamma}$ plus CM-stimulated peritoneal rnacrophages was decreased by the treatment with $N^{G}-monomethyl-{_L}-arginine{\;}(N^{G}MMA){\;}N^{\alpha}-Tosyl-Phe$ chloromethyl ketone (TPCK) , and was almost completely inhibited by pre-treatment with PDTC. Furthermore, treatment with CM alone or rIFN-{\gamma} plus CM in peritoneal macrophages caused a significant increase in $TNF-{\alpha}$ production. PDTC decreased CM-induced $TNF-{\alpha}$ production significantly. After CM treatment in HT-29 or AGS cells, cell viability decreased. Conclusions : These findings demonstrate that CM increases the production of NO and $TNF-{\alpha}{\;}by{\;}rIFN-{\gamma}-primed$ macrophages and suggest that NF-B plays a critical role in mediating these effects of CM.

• Journal of Microbiology and Biotechnology
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• 제28권10호
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• pp.1626-1634
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• 2018

Ginseng (the root of Panax ginseng Meyer) fermented by Lactobacillus plantarum has been found to attenuate allergic responses in in vitro and in vivo experimental models. Ginseng has been reported to also possess various biological functions including anti-inflammatory activity. The present study was aimed at comparing the anti-allergic effect of ginseng and fermented ginseng extracts on IgE-mediated passive cutaneous anaphylaxis in vitro in a murine cell line and in vivo in mice. Fermented ginseng extract (FPG) showed higher inhibitory effect against in vitro and in vivo allergic responses when compared with ginseng extract (PG). The secretion of ${\beta}$-hexosaminidase and interleukin (IL)-4 from the IgE-DNP-stimulated RBH-2H3 mast cells were significantly (p < 0.05) inhibited by FPG treatment, and this effect was concentration-dependent. Further, MKK4 activation and subsequent JNK phosphorylation were attenuated by FPG treatment. The inhibitory effect of FPG on the in vitro allergic response was verified in vivo against IgE-DNP-induced passive cutaneous anaphylaxis in a mouse model. These data indicated that the fermentation of ginseng with L. plantarum enhanced its anti-allergic effects both in vitro and in vivo. We predict that compositional changes in the ginsenosides caused by the fermentation may contribute to the change in the anti-allergic effects of ginseng. The results of our study highlight the potential of the use of FPG as a potential anti-allergic agent.

• Natural Product Sciences
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• 제18권2호
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• pp.130-136
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• 2012

In the present work, we have collected 19 species of macroalgae (9 Phaeophta and 10 Rhodophyta) f rom all around of Korea: Dictyopteris divaricata, D. prolifera, Myelophycus cavus, Papenfussiella kuromo, Petalonia zosterifolia, Petrospongium rugosum, Rugulopteryx okamurae, Sargassum fulvellum, S. muticum, Callophyllis japonica, Gloiopeltis tenax, Gracilaria longissima, Gracilaria vermiculophylla, Grateloupia asiatica, Grateloupia lanceolata, Grateloupia sparsa, Grateloupia turuturu, Grateloupia sp, and Polyopes affinis. The macroalgal species were extracted by 70% ethanol (EtOH) for 24 h and evaluated its inhibitory effects on ${\alpha}$-glucosidase. Among ethanol extracts, Myelophycus cavus showed the most effectively inhibitory activity ($IC_{50}$, 2.17 ${\mu}g/ml$) against ${\alpha}$-glucosidase, followed by Sargassum fulvellum (<$IC_{50}$, 8.13 ${\mu}g/ml$), Dictyopteris prolifera ($IC_{50}$, 16.66 ${\mu}g/ml$), Rugulopteryx okamurae ($IC_{50}$, 50.63 ${\mu}g/ml$), and Petrospongium rugosum ($IC_{50}$, 101.62 ${\mu}g/ml$). Furthermore, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay showed no cytotoxicity on mouse pre-adipocytes cell line (3T3-L1). These results suggest that some edible macroalgae merit further evaluation for clinical usefulness as anti-diabetic functional foods.

• Journal of Pharmaceutical Investigation
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• 제40권2호
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• pp.91-100
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• 2010

The objective of this work is to study transdermal delivery of donepezil hydrochloride (DH) using iontophoresis and to evaluate various factors which affect the transdermal transport. After the flux study using 4 kinds of hydrogel, hydrogel containing 8% poly(ethylene oxide) (PEO) was chosen as the hydrogel for further studies. Under experimental condition, DH was stable. We have studied the effect of polarity, current density, drug concentration and current profile on transdermal flux and compared the results. In vitro flux study was performed at $33^{\circ}C$, using side-by-side diffusion cell and full thickness hairless mouse skin. DH is positively charged at pH 7.4, and anodal delivery was much larger than cathodal and passive delivery at all current densities studied (0.2, 0.4 and 0.6 mA/$cm^2$). Cathodal delivery showed higher flux than passive flux. Flux increased as the concentration of DH in hydrogel increased. Pulsatile application of current showed smaller flux value than the application of continuous current. Based on these results, we have evaluated the possibility of delivering enough amount of DH to reach the therapeutic level. The maximum cumulative amount of DH transported for 12 hours was 455 ${\mu}g/cm^2{\cdot}hr$ when the amount of DH in the hydrogel was 3 mg/mL and the current density was 0.4 mA/$cm^2$. If the patch size is 10 $cm^2$, then we can deliver 4.6 mg for 12 hours. Because the daily dosage of DH is 5 mg, it seems possible to deliver clinically effective amount of DH using iontophoresis. This study also provides some information about the role of electrorepulsion and electroosmosis during the transport through skin.

• Archives of Pharmacal Research
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• 제21권5호
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• pp.503-507
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• 1998

Percutaneous absorption and model membrane variations of melationin (MT) in aqueous-based propylene glycol and $2-hydroxypropyl-{\beta}-cyclodextrin $vehicles were investigatted. the excised hairless mouse skin (HMS) and two synthetic ethylene vinyl acetate (EVA) and microporous polyethylene (MPE) were selected as a model membrane. the solubility of MT was determined by phase equilibrium study. the vertical $Franz{\circledR}$ type cell was used for diffusion study. The concentration of MT was determined using reverse phse HPLC system. The MT solubility was the highest in a mixture of PG and $2-HP{\beta}CD$. The percutaneous absorption of MT through excised HMS increased as the solubility increased. However, the permeability coefficient decreased and then slightly increased in mixture of PG and $2-HP{\beta}CD$. On the other hand, both flux and permeability coefficient through EVA membrane decreased as the solubility increased. No MT was detected over 12 h after starting diffusion through MPE membrane. The flux of MT was dependent on the type of membrane selected. Flux of MT was greatest in excised HMS followed by EBA and MPE membrane. Flux of MT through EVA membrane was 5-20 times lower when compared to excised HMS. Interestingly, volumes of donor phase when MPE membrane was used, significantly increased during the study period. the HMS might be applicable to expect plasma concentration of MT in human subjects based on flux and pharmacokinetic parameters as studied previously. the current studies may be applied to deliver MT transdermally using aqueous-based vehicles and to fabricate MT dosage forms.

• Archives of Pharmacal Research
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• 제25권2호
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• pp.191-196
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• 2002

The water extract from the root bark of Cortex Mori (CM, Morus alba L.: Sangbaikpi), a mulberry tree, has been known in Chinese traditional medicine to have antiphlogistic, diuretic, and expectorant properties. In this study, the cytotoxicity of CM against tumor cells and its mechanism was examined . CM exhibited cytotoxic activity on K-562, B38O human leukemia cells and B16 mouse melanoma cells at concentrations of > 1 mg/ml. A DNA fragmentation, PARP cleavage, and nuclear condensation assay showed that those cells exposed to CM underwent apoptosis. The water extract of Scutellarie Radix (SR) was used as a negative control and showed no cytotoxicity in those cells. The flow cytometric profiles of the CM-treated cells were also indicative of apoptosis. However, they did not appear to exert the G1 arrest, which is observed in other tubulin inhibitor agents such as vincristine, taxol. The protein-binding test using Biacore and a microtubule assembly-disassembly assay provided evidence showing that CM bound to the tubulins resulting in 3 markets inhibition of the assembly, but not the disassembly of microtubules. The possible nonspecific effect of the CM extract could be excluded due to the results using SR, which did not affect the assembly process. Overall, the water extract of CM induces apoptosis of tumor cells by inhibiting microtubule assembly.

• Biomolecules & Therapeutics
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• 제28권5호
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• pp.414-422
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• 2020

Fangchinoline (FAN) is a bisbenzylisoquinoline alkaloid that is widely known for its anti-tumor properties. The goal of this study is to examine the effects of FAN on arthritis and the possible pathways it acts on. Human fibroblast-like synovial cells (FLS), carrageenan/kaolin arthritis rat model (C/K), and collagen-induced arthritis (CIA) mice model were used to establish the efficiency of FAN in arthritis. Human FLS cells were treated with FAN (1, 2.5, 5, 10 µM) 1 h before IL-1β (10 ng/mL) stimulation. Cell viability, reactive oxygen species measurement, and western blot analysis of inflammatory mediators and the MAPK and NF-κB pathways were performed. In the animal models, after induction of arthritis, the rodents were given 10 and 30 mg/kg of FAN orally 1 h before conducting behavioral experiments such as weight distribution ratio, knee thickness measurement, squeaking score, body weight measurement, paw volume measurement, and arthritis index measurement. Rodent knee joints were also analyzed histologically through H&E staining and safranin staining. FAN decreased the production of inflammatory cytokines and ROS in human FLS cells as well as the phosphorylation of the MAPK pathway and NF-κB pathway in human FLS cells. The behavioral parameters in the C/K rat model and CIA mouse model and inflammatory signs in the histological analysis were found to be ameliorated in FAN-treated groups. Cartilage degradation in CIA mice knee joints were shown to have been suppressed by FAN. These findings suggest that fangchinoline has the potential to be a therapeutic source for the treatment of rheumatoid arthritis.

• 고려인삼학회:학술대회논문집
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• 고려인삼학회 2002년도 학술대회지
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• pp.522-530
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• 2002

We have already known, neural progenitor cells exist not only in the developing brain, but in certain spots in adult CNS in mammals, so it will be of great value to find out some compounds which can interfere these cells proliferation ability. In this research, we observed that ginsenoside $Rg_1$ can not only enhance neural progenitors' proliferation ability in vitro, but increase neurogenesis in adult mouse dentate gyrus in vivo. Firstly, we set up neural progenitor cells' culture system from embryonic rats' hippocampus and prove their feature through immunocytochemistry. Then by using MTT assay, we found that when growing with ginsenoside $Rg_1(0.5\~2.5{\mu}mol/l)$, the progenitor cells' survival rate nearly doubled, furthermore, we proved that this increase was due to the increment of cell proliferation through $^3H-thimidine$ incorporation assay, hence, we drew the first conclusion: ginsenoside Rg1 has the ability to stimulate neural progenitor cells' proliferation in vitro; in order to observe this compound's effect in vivo, we devised the following experiment: after administering ginsenoside Rg1 (5, 10 mg/kg, once a day) intraperitoneally for two weeks, we examine the number of BrdU positive cells in the dentate gyrus of mice, and found that Rg1 could increase the number of proliferation cells significantly in vivo. From these studies, we are quite sure about Rg1's effects on the proliferation ability of neural progenitor cells both in vitro and in vivo, certain targets of the compound and its underlying mechanisms are in progress.

• Nutrition Research and Practice
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• 제13권5호
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• pp.367-376
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• 2019

BACKGROUND/OBJECTIVE: The blue honeysuckle berry (Lonicera caerulea var. edulis L.) is a small deciduous shrub belonging to the Caprifoliaceae family that is native to Russia, China, Japan, and Korea. The berry of this shrub is edible, sweet and juicy and is commonly known as the blue honeyberry (BHB). This study examined the anti-diabetic potential of BHB on high-fat-diet-induced mild diabetic mice. The hypoglycemic, and nephroprotective effects of the 12-week oral administration of blue honeyberry extract were analyzed. MATERIALS/METHODS: The hypoglycemic effects were based on the observed changes in insulin, blood glucose, and glycated hemoglobin (HbA1c). Furthermore, the changes in the weight of the pancreas, including its histopathology and immunohistochemical investigation were also performed. Moreover, the nephroprotective effects were analyzed by observing the changes in kidney weight, its histopathology, blood urea nitrogen (BUN), and serum creatinine levels. RESULTS: The results showed that the high-fat diet (HFD)-induced control mice showed a noticeable increase in blood glucose, insulin, HbA1c, BUN, and creatinine levels. Furthermore, growth was observed in lipid droplet deposition related to the degenerative lesions in the vacuolated renal tubules with the evident enlargement and hyperplasia of the pancreatic islets. In addition, in the endocrine pancreas, there was an increase in the insulin-and glucagon-producing cells, as well as in the insulin/glucagon cell ratios. On the other hand, compared to the HFD-treated mice group, all these diabetic and related complications were ameliorated significantly in a dose-dependent manner after 84 days of the continuous oral administration of BHBe at 400, 200 and 100 mg/kg, and a dramatic resettlement in the hepatic glucose-regulating enzyme activities was observed. CONCLUSIONS: By assessing the key parameters for T2DM, the present study showed that the BHBe could act as a potential herbal agent to cure diabetes (type II) and associated ailments in HFD-induced mice.