Munoz-Perez, Victor Manuel;Fernandez-Martinez, Eduardo;Ponce-Monter, Hector;Ortiz, Mario I.
The Korean Journal of Physiology and Pharmacology
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v.21
no.4
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pp.429-437
/
2017
The aim of this study was to evaluate the relaxant and anti-inflammatory effects of two thalidomide analogs as phosphodiesterase-4 (PDE-4) inhibitors in pregnant rat uterus. Uteri from Wistar female rats were isolated at 19 day of pregnancy. Uterine samples were used in functional studies to evaluate the inhibitory effects of the thalidomide analogs, methyl 3-(4-nitrophthalimido)-3-(3,4- dimethoxyphenyl)-propanoate (4NO2PDPMe) and methyl 3-(4-aminophthalimido)- 3-(3,4-dimethoxyphenyl)-propanoate (4APDPMe), on prostaglandin-$F2{\alpha}$ ($PGF2{\alpha}$)-induced phasic, $K^+$-induced tonic, and $Ca^{2+}$-induced contractions. Accumulation of cAMP was quantified in uterine homogenates by ELISA. Anti-inflammatory effect was assessed by using ELISA for determination of the pro-inflammatory cytokines tumor necrosis factor-${\alpha}$ ($TNF{\alpha}$) and interleukin (IL)-$1{\beta}$, and anti-inflammatory IL-10, from uterine explants stimulated with lipopolysaccharide (LPS). Nifedipine, forskolin and rolipram were used as positive controls where required. Both thalidomide analogs induced a significant inhibition of the uterine contractions induced by the pharmaco- and electro-mechanic stimuli. Nifedipine and forskolin were more potent than the analogs to inhibit the uterine contractility, but these were more potent than rolipram, and 4APDPMe was equieffective to nifedipine. Thalidomide analogs increased uterine cAMP-levels in a concentration-dependent manner. The LPS-induced $TNF{\alpha}$ and $IL-1{\beta}$ uterine secretion was diminished in a concentration-dependent fashion by both analogs, whereas IL-10 secretion was increased significantly. The thalidomide analogs induced utero-relaxant and anti-inflammatory effects, which were associated with the increased cAMP levels as PDE-4 inhibitors in the pregnant rat uterus. Such properties place these thalidomide analogs as potentially safe and effective tocolytic agents in a field that urgently needs improved pharmacological treatments, as in cases of preterm labor.
Song, Ji Hyun;Lee, Jin Hwa;Kim, Eun Jin;Kim, Yun Hee
The Journal of Pediatrics of Korean Medicine
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v.32
no.3
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pp.1-15
/
2018
Objectives Alismatis Rhizoma has been known to suppress inflammation and allergic reaction. However, the cellular target of Alismatis Rhizoma and its mechanism of action remain unclear. This study was designed to examine the effect of Alismatis Rhizoma extract (ALC) on the RBL-2H3 mast cells in vitro and on the OVA/alum sensitized mice ex vivo. Methods In the study, RBL-2H3 mast cells were cultured in minimal essential medium (MEM) for 24 hours, and treated separately with cyclosporin A and varying doses of ALC, and then stimulated with Phorbol 12-myristate 13-acetate (PMA) (50 ng/ml) and Ionomycin ($0.5{\mu}M$). The levels of IL-13, IL-4 were measured by ELISA analysis. The mRNA levels of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$ were analyzed with Real-time PCR. Also, manifestations of MAPKs transcription factors and $NF-{\kappa}B$ p65 translocation were analyzed by western blotting in vitro. Subsequently, for ex vivo experiment, we induced allergic inflammation on Balb/c mice by OVA/alum and administered ALC orally. And we measured serum OVA-specific IgE level and IL-4, IL-13 in the splenocyte culture supernatant by ELISA analysis. Results ALC was shown to suppress mRNA expression of IL-4, IL-5, IL-6, IL-13, GM-CSF, $TNF-{\alpha}$, and to inhibit the IL-13, IL-4 production. Also ALC reduced an activation of mast cells specific signal MAPKs transcription factors and $NF-{\kappa}B$ p65 from the western blot analysis in in vitro experiment. In ex vivo, ALC oral adminstration decreased the level of OVA-specific IgE in serum, and IL-4, IL-13 in the splenocyte culture supernatant. Conclusions ALC is shown to reduce inflammation and allergic response by suppressing Th2 cytokines through the regulation of transcription factors MAPKs and $NF-{\kappa}B$ p65 in mast cells. Administration of ALC suppressed OVA-specific IgE in ovalbumin allergy model through the inhibition of Th2 cytokine. In conclusion, ALC can be considered as an effective treatment for allergic diseases such as atopic dermatitis.
Le, Thanh Hoa;Le, Tran Binh;Doan, Thanh Huong Thi;Quyen, Dong Van;Le, Kim XuyenThi;Pham, Viet Cuong;Nagataki, Mitsuru;Nomura, Haruka;Ikeue, Yasunori;Watanabe, Yoshiya;Agatsuma, Takeshi
Journal of Microbiology and Biotechnology
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v.21
no.4
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pp.405-411
/
2011
Avian influenza virus vaccines produced in oil-emulsified inactivated form with antigen content of at least 160 hemagglutinin units (HAU) induced immunity in birds. However, in addition to enhancing the effect of the adjuvant(s), other additional supplemented biological compounds included in inactivated vaccines could produce higher levels of antibody. We examined in chickens, Vietnamese ducks, and muscovy ducks the adjuvant effect of Sophy ${\beta}$-glucan (SBG), a ${\beta}$-1,3-1,6 glucan produced by the black yeast Aureobasidium pollulans strain AF0-202, when administered with an avian influenza H5 subtype vaccine. In Experiment 1, 40 chickens (ISA Brown hybrid), allocated to four groups of ten each, were immunized with Oil-H5N1(VN), Oil-H5N1(CN), Oil-H5N2(CN), and saline (control group), respectively. In Experiment 2, chickens (ISA Brown hybrid), muscovy ducks (French hybrid), and Vietnamese ducks (indigenous Vietnamese) were used to further assess the effect of SBG on immunogenicity of the Oil-H5N1(VN) Vietnamese vaccine. ELISA and hemagglutination inhibition (HI) assays were used to assess the antibody response. The H5 subtype vaccines initiated significantly higher immune responses in the animals dosed with SBG, with 1.0-1.5 $log_2$ higher HI titers and 10-20% ELISA seroconversion, compared with those not dosed with ${\beta}$-glucan. Notably, some of the animals dosed with SBG induced HI titers higher than 9.0 $log_2$ following boosting immunization. Taken together, our serial studies indicated that SBG is a potential effector, such as enhancing the immune response to the H5 vaccines tested.
The effects of the extracts from the bran part of pigmented rices on inflammation was evaluated by determining their inhibitory action on the histamine and ${\beta}-hexosaminidase$ release, together with inflammatory cytokine productions ($IL-1{\beta},\;TNF-{\alpha}$ and IL-6). Examination of the inhibitory effects on the histamine and ${\beta}-hexosaminidase$ release from a basophilic cell line RBL-2H3 cells and rat peritoneal mast cells (RPMC) showed that the pigmented rice extract inhibited these inflammation-mediating substances (10.19% and 110.03% inhibition in histamine and ${\beta}-hexosaminidase$ release, respectively), while normal brown rice extract rather increased their release. For RPMC, the pigmented rice extract was found to have 8 or 3-fold stronger inhibitory activity than normal brown rice toward histamine or ${\beta}-hexosaminidase$ release, respectively. Expression of $IL-1{\beta},\;TNF-{\alpha}$ and IL-6 was measured as the representative inflammatory cytokine species showed that the pigmented rice extract had a higher inhibitory activity than the normal rice counterpart. ELISA analysis for determining cytokine release demonstrated a more effective blockading ability of the pigmented rice to the release of $IL-{\beta},\;TNF-{\alpha}$ and IL-6 compared to normal rice. These results showed us the superiority of the pigmented rice bran extract not only in suppressing the release of inflammation-mediating substances such as histamine and ${\beta}-hexosaminidase$, but also in repression of the inflammatory cytokine expression.
Journal of the Society of Cosmetic Scientists of Korea
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v.32
no.3
s.58
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pp.135-140
/
2006
Nuclear factor-kappaB (NF-kappaB) is a critical transcription factor for maximal expression of many of the cytokines that are involved in the pathogenesis of inflammatory diseases. In this study, we found that 12 plant extracts among 200 plants, namely, Forsythia koreana, Capsicum annuum L, Mentha arvenis, Duchesnea chrysantha, Morus alba, Saururus Chinenis (Lour) Baill, Pine needle, Zingiber mioga (Thunb.), Roscoe, Houttuynia, Prunus yedoenis, Sasa quelpaertenis, significantly inhibited LPS- induced NF-kappaB activation in a concentration-dependent manner. Additionally, 12 plant extracts were found to have antioxidant activities in DPPH assay Therefore, we have attempted to determine whether 12 herbal extracts could inhibit the expression of cytokines possessing NF-kappaB promoter in their promoter regions. Consistently 12 herbal extracts inhibited LPS-induced production of TNF alpha and interleukin-8 (IL-8). These results show that 12 herbal extracts suppresses the production of pro-inflammatory mediators through the inhibition of the NF-kappaB signaling pathway, we suggest that 12 herbal extracts can be used as a anti-inflammatory and soothing agent.
Periodontal therapy for treatment of periodontitis involves the elimination of bacterial plaque and elimination of the anatomic defects by regenerative procedure. The purpose of this study was to evaluate on the biological effect of magnolia and Ginkgo biloba extract to the antimicrobial, antiinflammatory and cellular activity. Antimicrobial assay was performed with the diffusion method of the extract by measuring of growth inhibitory zone of B. cereus from blood agar plate. Effect of the extract to cellular activity of gingival fibroblast were examined using MTT method and measured the result with optical density on 570nm by ELISA reader. Inhibitory effects of $PGE_2$ production from gingival fibroblast was performed with the addition of $IL-l{\beta}$ and the extract to the well and examined to the product of $PGE_2$ from cell by ELISA reader. In vivo anti-inflammatory effect was performed with injection examined with clinically and histologically for their extent of mecrosis and inflammation. Antimicrobial activity of Magnolia extract showed significantly higher activity than that of control. However, GBE did not showed significant activity to compare with control, and mixture of Magnolia and GBE extract showed significantly higher activity than that of control. The effect of cellular activity to gingival fibroblast showed no significant differences of between control and Magnolia extract. However, GBE showed significantly higher rate of cellular activity to compare with control and even to PDGF-BB, and also showed same degree of cellular activity even though mixed with Magnolia extract. The inhibitory effect of $PGE_2$ production showed significantly reduction of $PGE_2$ production to compare with control, but its inhibitory effect was not much strong to compare with Indomethacin. In vivo, antiinflammatory effect of Magnolia extract to P. gingivalis injection of Hamster buccal check showed significantly reduction of inflammatory cell infiltration and tissue necrosis, but GBE showed no effect on the inhibition of inflammatory process. These results suggested that Magnolia and GBE extract possessed different kind of biological activity and also can be compensated on their activity with each other for elimination of bacterial plaque and anatonical defect.
Park, Bo-Hyoung;Yoo, Seung-Ah;Hong, Kyung-Hee;Hyoung, Bok-Jin;Hwang, Yu-Na;Cho, Chul-Soo;Park, Won;Kim, Wan-Uk
IMMUNE NETWORK
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v.6
no.1
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pp.33-42
/
2006
Background: Calcineurin plays a crucial role in T cell activation, cell growth, apoptosis, and angiogenesis, and its over-expression has been implicated in the pathogenesis of cardiomyopathy and stroke. However, the expression and function of calcineurin in the pathologic lesion of chronic inflammatory diseases, like rheumatoid synovium, remain to be defined. This study was aimed to determine the role of calcineurin in inflammatory arthritis and investigate the expression and function of calcineurin in the rheumatoid synovium and synoviocytes, the actual site of chronic inflammation. Methods: Immuno-histochemical staining using specific antibody to calcineurin was perfomed in the synovium of rheumatoid arthritis (RA). Fibroblast-like synoviocytes (FLS) from RA and osteoarthritis (OA) patients were isolated from RA and OA patients, and cultured with IL-1${\beta}$ and TNF-${\alpha}$ in the presence or absence of cyclosporin A, a calcineurin inhibitor. The calcineurin expression was assessed by phosphatase assay and Western blotting analysis. IL-6, -10, -17, matrix metalloproteinase (MMP)-1, -2, -3, and -9 released into the culture supernatants were measured by ELISA. After transfection with GFP-Cabin 1 gene into synoviocytes, the levels of IL-6 and MMPs were measured by ELISA. Results: Calcineurin was highly expressed in the lining layer of synovium and cultured synoviocytes of RA patients. The elevated calcineurin activity in the rheumatoid synoviocytes was triggered by proin flammatory cytokines such as IL-1${\beta}$ and TNF-${\alpha}$. In contrast, IL-10, an anti-inflammatory cytokine, failed to increase the calcineurin activity. The targeted inhibition of calcineurin by the over-expression of Cabin 1, a natural calcineurin antagonist, inhibited the production of IL-6 and MMP-2 by rheumatoid synoviocytes in a similar manner to the calcineurin inhibitor, cyclosporin A. Conclusion: These data suggest that abnormal activation of calcineurin in the synoviocytes may contribute to the pathogenesis of chronic arthritis, and thus provide a potential target for controlling inflammatory arthritis.
Sin, Mi-Kyung;Kil, Ki-Jeong;Lee, Young-Chul;Kim, Jin-Suk;Seo, Young-Bae;Roh, Seong-Soo
The Korea Journal of Herbology
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v.20
no.2
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pp.159-169
/
2005
Objectives : This study was carried out for the purpose of knowing the effect from anti-arthma action of the abstraction from a extract of Paridis Rhizoma(EPR). In order to know what the effect of controlling an abstraction from Paridis Rhizoma. and about the expression of B cells and Ig E cells, mast cells it was necessary for it to be activated by ovalbumin. Methods : In order to know what the effect was on the organization of cytokine gene expression from The increase and divorce of the B cells and allergic acting by EPR, we found it necessary to examine the BALF. At the same time, as we examined the histamine release by ELISA method, we also examined the effect of EPR. Results : EPR at $100\;{\mu}g/ml$, the highest concentration examined did not have any cytotoxic effects on mLFCs. In FACS analysis, number of granulocyte/lymphocyte, $CD3e^+/CCR3^+,\;CD4^+\;and\;CD23^+/B220^+$ in asthma-induced lung cells were significantly decreased by EPR treatment compared to the control group. In RT-PCR analysis, mRNA expression for CCR3, eotaxin and histamine in asthma-induced lung cells, which was induced by rIL-3 plus rmIL-5 treatments, was significantly decreased by EPR treatment. In ELISA analysis, production levels of IL-4, IL-13 and histamine in asthma-induced lung cells, which were induced by rIL-3 plus rmIL-5 co-treatment, were significantly decreased by EPR treatment. EPR treatments significantly inhibited the proliferation of eosinohils prepared from asthma-induced mouse lung tissues compared to the non-EPR treated control cells. Immunohistochemical analysis revealed that EPR treatment significantly decreased the levels of eosipnphil activation compared to non-treated cells. Conclusion : The present data suggested that Paridis Rhizoma may have an effects on the inhibition of parameters associated with asthma responses in eosinpophils, and thus implicate the possibility for the clinical application of Paridis Rhizoma.
Mushroom-derived ${\beta}$-glucan, a polysaccharide (GLP) isolated from the mycelium of Ganoderma lucidum, was previously shown to have inhibitory effects against tumor-bearing mice in vivo. We investigated the apoptotic effect of mushroom-derived ${\beta}$-glucan in a sarcoma-180 tumor cell- bearing mice model using an ELISA to determine the levels of tumor necrosis factor-${\alpha}$ (TNF-${\alpha}$) in the mice. The morphology of the tumor cells was assessed with transmission electron microscopy (TEM). GLP was injected into the tumor-bearing mice at a dose (i.p.) of 20 mg/kg for 10 days. After 30 days, the tumor mass from the inguinal region was collected, weighed, and assayed using TEM and a TNF-${\alpha}$ ELISA kit. The tumors that developed in the mice treated with GLP were 71.4% smaller than those in the control group, showing the ability of GLP to inhibit tumor growth. The levels of TNF-${\alpha}$ in the serum of the sarcoma-180 bearing mice were 12 times greater than in the serum of the nonbearing tumor mice. An ultrastructural study demonstrated that the GLP-treated sarcoma-180 tumor cells were condensed, with rearranged chromatin. In addition, the marginated chromatin in nucleus induced the nuclear compartment, and there were many vacuolization in the cell. GLP could be an effective apoptosis-inducing compound in sarcoma-type cancers.
Dong-hwan, Lee;Jin-hwa , Kim;Jun-tae, Bae;Sung-min, Park;Hyeong-bae, Pyo;Tae-boo, Choe;Bum-chun, Lee
Journal of the Society of Cosmetic Scientists of Korea
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v.30
no.2
/
pp.241-246
/
2004
To develop novel anti-aging peptides from the germinated black rice, we treated with bromelain, papain and Pronase E. And we investigated the effects of the germinated black rice peptide (GBRP) as anti-aging cosmetic ingredients, and compared with the non-germinated black rice protein (NBRP). We investigated the effects on in vitro inhibition of matrix-metalloprotease (MMP), proliferation of human skin fibroblasts, stimulation of collagen synthesis and expression of UVA-induced MMPs in human skin fibroblasts, UVA induced MMP-1 expression and collagen contents in human skin fibroblasts were analyzed by enzyme-linked immunosorbent assay (ELISA). As a result, the molecular weight distributions of GBRP and NBRP were determined by gel permeation chromatography to be approximately 900 and 10,000 daltons. GBRP increased skin cell proliferation about 40% and reduced UVA-induced MMP-1 expression about 50%. Also the collagen protein level of cells, which were cultured with GBRP, was increased about 25%. These results suggest that the geminated plant seed peptides can be novel anti-aging ingredients for cosmetics.
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