Impact of compound 48/80 activated mast cell on dendritic cell homing to lymph nodes / 中国免疫学杂志

Objective:To investigate the effect of compound 48/80(C48/80) activated mast cell on dendritic cell homing to draining lymph nodes(DLN) in order to explore the potential mechanism of C48/80 promoting adaptive immune response.Methods: C57BL/6 mice bone marrow derived dendritic cells(BM-DC) and MC/9 mast cell line were propagated in vitro.Chemotaxis of BM-DC supplemented with supernatant of C48/80 treated MC/9 to CCL21 was measured by transwell chemotaxis assay.C48/80 was injected into murine scapular site,local skin mast cell degranulation was detected by toluidine blue staining method.The number of CD11c+ DC in draining lymph nodes(DLN) was detected by Flow cytometry.Exogenous green microbeads labeled BM-DC homing to DLN was detected by fluromicroscope.The mice pretreated with or without C48/80 were vaccinated by ovalbumin pulsed DC(OVA-DC),DLN lymphocyte proliferation was detected by WST-8 reagent.Results: A large amount of typical BM-DC were harvested from media supplemented with murine recombinant IL-4 and GM-CSF.Product of activated MC/9 enhanced chemotaxis of BM-DC to CCL21(P<0.01).Intradermal injection of C48/80 induced local mast cell obvious degranulation and local inflammation.Mice pretreated with C48/80 demonstrated high number of total cells and DC cells in DLN.The number of fluorescent positive BM-DC in DLN also increased in C48/80 pretreated mice.Antigen specific lymphocyte proliferation enhanced in OVA-DC inoculated C48/80 pretreated mice.Conclusion: Mast cell activation induced by C48/80 can enhance DC homing to DLN and increase specific lymphocyte proliferation,which may be one of a mechanism of C48/80 in promoting adaptive immune response.

Effect of conjugated linoleic acid on compound 48/80-induced pruritus in mice / 동물의과학연구지

Pruritus is one of the most important symptoms of allergic inflammatory skin disease. Conjugated linoleic acid (CLA) has been reported to have preventive effects against allergic inflammation. The objective of this study was to determine whether or not oral administration of CLA suppresses pruritus induced by compound 48/80 (composed of N-methyl-p-methoxy phenethylamine with formaldehyde) in mice, and if so, whether or not this effect is associated with serum histamine and prostaglandin (PG) E2 levels. Liquid CLA mixture (36.25% 9c-11t CLA, 36.95% 10t-12c CLA, 1.12% 9c-11c, and 1.94% t9-t11 CLA) was emulsified in 0.5% carboxymethyl cellulose (CMC) sodium salt and orally administered to mice at doses of 200 mg/kg once per day for 3 days. Similarly, disodium chromoglycate (DSCG), an antipruritic substance, was administered orally at the same concentrations as the negative control. Compound 48/80, a pruritus-inducing reagent, was subcutaneously injected 30 minutes after final administration of CLA. Scratching behavior of mice was counted just after compound 48/80 injection. Serum histamine and PGE2 concentrations were evaluated individually. Mice administered with CLA showed reduced frequency of scratching behavior compared to those without CLA. Antipruritic activities in CLA-treated and DSCG-treated groups were 48.5% and 26.8%, respectively. CLA and DSCG also diminished serum concentrations of histamine and PGE2 compared to compound 48/80 alone, respectively. This result suggests that dietary CLA has an antipruritic effect by down-regulating serum histamine and PGE2 levels for relief of compound 48/80-induced scratching behavior in mice, which will be useful in allergic pruritus as a preventive medicine.

Evaluation of anti-allergic activity of gossypin and suramin in mast cell-mediated allergy model.

The mast cell-mediated allergic reactions are involved in many allergic diseases, such as asthma, allergic rhinitis and sinusitis. Stimulation of mast cells initiates the process of degranulation, resulting in the release of mediators such as histamine and an array of inflammatory cytokines. In this report, we investigated the effect of gossypin (a biflavonoid) and suramin (a synthetic polysulphonated naphtylurea) on the mast cell-mediated allergy model, and studied the possible mechanism of their action. Both gossypin and suramin inhibited (P<0.001) compound 48/80-induced systemic anaphylaxis reactions, antiprurities (P<0.001) and reduced the histamine release in rats. Further, both showed significant (P<0.001) protection against rat peritoneal mast cells activated by compound 48/80. Thus, our findings provide evidence that gossypin and suramin inhibit mast cell-derived allergic reactions.

Inhibitory Effect of Bear Bile on Compound 48/80-induced Mast Cell Activation and IgE-mediated Vascular Permeability / 체질인류학회지

The bear bile has been used as a traditional drug medicine and has been known to have anti-tumor, anti-inflammatory and anti-oxidant effects. The purpose of this study is to investigate the inhibitory effect of bear bile on compound 48/80-induced mast cell activation in vitro and anti-dinitrophenyl (DNP) IgE-mediated vascular permeability in vivo. For this, the effects of bear bile on the degranulation, histamine release, calcium influx and the change of the intracellular cAMP levels of rat peritoneal mast cells (RPMCs) and the influences of the oral treatment of bear bile on IgE-mediated cutaneous vascular permeability were studied. the results were as follows; the compound 48/80-induced degranulation, histamine release and calcium influx of RPMCs were inhibited by pretreatment with bear bile, the cAMP levels of RPMCs were increased by pretreatment with bear bile, and bear bile inhibited anti-DNP IgE-mediated cutaneous vascular permeability. From the above results, it is suggested that bear bile contains some substances which inhibit anti-DNP IgE-mediated vascular permeability and mast cell activation. Bear bile potentially may serve as an effective therapeutic agent for allergic diseases.

Inhibitory Effect of Arctium lappa Linne on Compound 48/80-induced Mast Cell Activation and Vascular Permeability / 체질인류학회지

Arctium lappa Linne (AL) has been widely cultivated for a long time as a popular vegetable. The fruit of AL has been used as an antiphlogistic and expectorant in herbal medicine, the crude drug is known as "burdock" in korea. In pharmaceutical field, a few papers recently reported the antiinflammatory, antiviral, and anticancer effects of this extract. However, the antiallergic effect of AL is unknown. The purpose of this study is to investigate the inhibitory effect of AL on compound 48/80-induced mast cell activation. For this, the effects of AL on the degranulation, the histamine release, and the change of the intracellular cAMP (cyclic adenosine-3, 5monophosphate) levels of rat peritoneal mast cells (RPMC) and the influences of AL on the compound 48/80-induced cutaneous reaction were studied. The results were as follows; 1) the compound 48/80-induced mast cell degranulation and histamine release of RPMC was significantly inhibited by pretreatment with AL, 2) the compound 48/80 decreased the cAMP levels of RPMC, but the compound 48/80-induced the cAMP levels of RPMC were significantly increased by pretreatment with AL, 4) AL significantly inhibited compound 48/80-induced vascular permeability of cutaneous tissue. From the above results, it is suggested that AL contains some substances which inhibit the compound 48/80-induced vascular permeability and mast cell activation.

Inhibitory Effect of Corni fructus on Compound 48/80-induced Mast Cell Activation and Vascular Permeability / 체질인류학회지

The fruit of Corni fructus (CF), a perennial herb, is believed to have anti-allergy effects, but its mechanism is unknown. The purpose of this study is to investigate the inhibitory effect of CF on compound 48/80-induced mast cell activation. For this, the effects of CF on the degranulation, the histamine release, the calcium influx and the change of the intracellular cAMP levels of rat peritoneal mast cells (RPMC) and influences of CF on the compound 48/80-induced cutaneous reaction were studied. The results were as follows; the compound 48/80-induced degranulation, intracelluar calcium influx and histamine release of RPMC was significantly inhibited by pretreatment with CF, the compound 48/80-induced cAMP level of RPMC were significantly increased by pretreatment with CF, CF significantly inhibited compound 48/80-induced vascular permeability of rat cutaneous tissue. From the above results, it is suggested that CF contains some substances which inhibit the compound 48/80-induced vascular permeability and mast cell activation.

Inhibitory Effect of Corni fructus on Compound 48/80-induced Mast Cell Activation and Vascular Permeability / 체질인류학회지

The fruit of Corni fructus (CF), a perennial herb, is believed to have anti-allergy effects, but its mechanism is unknown. The purpose of this study is to investigate the inhibitory effect of CF on compound 48/80-induced mast cell activation. For this, the effects of CF on the degranulation, the histamine release, the calcium influx and the change of the intracellular cAMP levels of rat peritoneal mast cells (RPMC) and influences of CF on the compound 48/80-induced cutaneous reaction were studied. The results were as follows; the compound 48/80-induced degranulation, intracelluar calcium influx and histamine release of RPMC was significantly inhibited by pretreatment with CF, the compound 48/80-induced cAMP level of RPMC were significantly increased by pretreatment with CF, CF significantly inhibited compound 48/80-induced vascular permeability of rat cutaneous tissue. From the above results, it is suggested that CF contains some substances which inhibit the compound 48/80-induced vascular permeability and mast cell activation.

Study on the Activation of Mast Cell by Seminal Plasma in vitro and in vivo (II : Effects of seminal plasma from the azoospermic, oligozoospermic and normozoospermic men on the degranulation and histamine release of rat peritoneal mast cells) / 체질인류학회지

Although it has been known that seminal plasma (SP) modulates immune responses, the morphologic and functional effects of SP on mast cells have not been well documented. The purpose of this study was to determine the morphologic and functional effects of SP from the azoospermic (ASP), oligozoospermic (OSP) and normozoospermic (NSP) men on rat peritoneal mast cells (RPMC). By inverted microscopy, degranulation of RPMC occurred by treatment of OSP, NSP or compound 48/80. Treatment of RPMC with OSP and NSP resulted in increase in size and decrease in surface folds and bulged granules from the cell surface indicating that degranulating processes took place. Irregular cell surface with few microvilli and large degranulation channels filled with altered granules of various sizes were observed in OSP or NSP-treated RMPC. The degranulation index (DI) of RPMC treated with HBSS was 8.8+/-5.0. SP-induced DI was 11.9+/-6.2, 57.1+/-16.9 and 61.9+/-15.8, in ASP, OSP and NSP, respectively. The DI of RPMC treated with compound 48/80 (1 microgram/ml) was 83.5+/-21.4. These results indicated that the OSP, NSP but not ASP, induced the degranulation of RPMC. OSP and NSP induced histamine release (HR) from RPMC (4.5x105 cells). The HR of RPMC treated with HBSS was 1,030+/-196.6ng/ml, whereas ASP, OSP and NSP-induced HR was 1,010+/-204.7, 2,794+/-453.3 and 2,899 +/-366.7 ng/ml, respectively. The HR of RPMC treated with compound 48/80 was 6,300+/-476.2 ng/ml. Intracellular calcium level(ICL) of RPMC was also increased by OSP and NSP. The ICL of RPMC treated with HBSS was 6.1+/-1.0 pmole. ASP, OSP and NSP-induced ICL was 9.0+/-1.1, 81.2+/-18.5 and 76.6+/-18.0 pmole, respectively. The ICL of RPMC treated with compound 48/80 was 102.9+/-22.2 pmole. From the above results, it is suggested that OSP and NSP contain some factors to degranulate RPMC by the increase of intracellular calcium level.

Inhibitory Effect of Polysaccharide Fraction from Cortex Mori on Compound 48/80-Induced Mast Cell Activation / 대한면역학회지

Cortex mori (Morus alba L.: Sangbaikpi), the root bark of mulberry tree, has been used as an antiphlogistic, diuretic, and expectorant in herbal medicine. Previous studies have demonstrated that the phenolic extract of Cortex mori have hypotensive, hypoglycemic, antifungal, antiviral, antiinflammatory, and anticancer effects, and the hot water extract from Cortex mori has inhibitory effects on compound 48/80- induced mast cell degranulation and histamine release from rat peritoneal mast cells (RPMCs). This study was perforrned to investigate the effects of polysaccharide fraction from Cortex mori (PFCM) on compound 48/80-induced degranulation, histamine release, calcium influx, changes of intracellular cAMP and cGMP level, and morphological changes of RPMCs. The results were summarized as follows. 1) Compound 48/80-induced cytomorphological changes such as swelling, degranulation, intracellular vacuoles, and interrupted cell boundary were significantly inhibited by pretreatment with either hot water or polysaccaride fractions frorn Cortex mori (PFCM), 2) the compound 48/80-induced histamine release from RPMCs pretreated with PFCM was significantly inhibited, compared to that of control without PFCM pretreatment, 3) the PFCM inhibited remarkably the compound 48/80-induced calcium influx into the RPMCs, 4) the PFCM increased significantly the intracellular cAMP levels and decreased the intracellular cGMP levels of RPMCs, compared to those of normal control, and 5) the compound 48/80-induced cAMP levels of RPMCs pretreated with PFCM were significantly increased, compared to those of positive control without PFCM, and the compound 48/80-induced cGMP levels of RPMCs pretreated with PFCM were remarkably decreased, compared to those of positive control without PFCM. From the above results, it is suggested that PFCM have an activity to inhibit the compound 48/80-induced mast cell activation.

The role of H1- and H2-receptors in the effect of compound 48/80 in the asphyxiation and body temperature of mice

Contribution of histamine H1- and H2-receptors to the effect of compound 48/80, a potent histamine releaser, upon asphyxiation and body temperature in mice was investigated in the present experiments. Compound 48/80 showed an apparent protective potency against hypoxia and significantly prolonged the latencies for convulsions and death in a dose-dependent manner. Compound 48/80 also decreased the body temperature, which was in relation with the antihypoxic effect. Both the H1-receptor antagonist, dimethindene, and the H2-receptor antagonist, ranitidine, attenuated the hypothermic effect of compound 48/80, indicating the involvement of central histamine through both the H1- and H2-receptors. Ranitidine had no effect on the protective effect of compound 48/80 against hypoxia-induced lethality, whereas dimethindene completely antagonized it. These results suggest that the protective effect of compound 48/80 against hypoxia is mediated through histamine H1-receptors and is not related to its ability to induce hypothermia.