Lactococcus lactis KR-050L extract suppresses house dust mite induced-atopic skin inflammation through inhibition of keratinocyte and mast cell activation.

AIMS: Atopic dermatitis (AD) is a chronic inflammatory skin disease, with a steadily increasing prevalence. Lactic acid bacteria (LAB) have been widely used in the food industry and are an attractive option for preventing and treating allergic skin diseases. We previously isolated new LABs including Lactococcus lactis KR-050L from Gajuknamu kimchi, and showed the anti-inflammatory effects of extract of L. lactis KR-050L culture broth (LLK). In this study, we investigated the effects of LLK on AD. METHODS AND RESULTS: For the in vitro study, we used human keratinocytes (HaCaT) and mast cells (RBL-2H3). In vivo study, we investigated the effects of LLK on Dermatophagoides farinae extract (DFE) and 2,4-dinitrochlorobenzene (DNCB)-induced atopic skin inflammation in mice. LLK suppressed expression of pro-inflammatory cytokines and chemokines by down-regulation of p38 MAPK, STAT1 and nuclear translocation of NF-κB in keratinocytes. Topical application of LLK suppressed AD symptoms based on reduction in ear thickness, serum IgE levels and immune cell infiltration. Furthermore, LLK inhibited serum histamine levels and mast cells infiltration in vivo, and reduced mast cells activation in vitro. CONCLUSIONS: These results suggest that LLK inhibits AD symptoms through inhibition of keratinocytes and mast cells activation. SIGNIFICANCE AND IMPACT OF THE STUDY: LLK is a potential therapeutic candidate for AD treatment.

Development of porcine oocytes from preovulatory follicles of different sizes after maturation in media supplemented with follicular fluids.

The development of porcine oocytes from large (3.1-8.0 mm in diameter) or small (<3.1 mm) follicles was examined after maturation culture in medium containing porcine follicular fluid (pFF). Large follicles yielded larger (256 microm v. 221 microm; P<0.05) cumulus-oocyte complexes and more (22 v. 14%) morphologically normal oocytes than small follicles (Experiment 1). In Experiments 2-4, maturation media supplemented with mixed pFF (10%) from small and large follicles was used. More oocytes from large follicles matured (58% v. 91%), formed pronuclei (81% v. 90%) and developed to the blastocyst stage (2% v. 10%) than oocytes from small follicles. In Experiments 5-7, the effects of pFF collected from either small or large follicles on oocyte development were examined. Regardless of the source of oocytes, large-follicle-derived pFF more significantly enhanced preimplantation development than did small-follicle-derived pFF. The highest rate of blastocyst formation (16%) was found when oocytes from large follicles were cultured in maturation medium containing large-follicle-derived pFF. These results suggest that oocytes from large follicles have greater developmental potential than oocytes from small follicles, and that the origin of pFF, which is added to the maturation media, might be an important factor for improving in vitro development of porcine oocytes.

Effect of Cryptotympana atrata extract on compound 48/80-induced anaphylactic reactions.

We studied the effect of the aqueous extract of Cryptotympana atrata Fabricius on anaphylactic reactions. Cryptotympana atrata completely inhibited compound 48/80-induced systemic anaphylactic shock at doses of 0.5 and 1.0 g/kg. When Cryptotympana atrata was given as pretreatment at concentrations ranging from 0.025 to 1.0 g/kg, the plasma histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. Cryptotympana atrata also inhibited histamine release from rat peritoneal mast cells (RPMC) by compound 48/80. In addition, when Cryptotympana atrata was added, the level of cAMP in RPMC transiently and significantly increased about 14-fold compared with that of control cells. Our studies provide evidence that Cryptotympana atrata may be beneficial in the treatment of nonspecific anaphylactic reactions.

Antianaphylactic activity of Poncirus trifoliata fruit extract.

The effect of an aqueous extract of Poncirus trifoliata (L.) Raf. (Rutaceae) fruits (PTFE) on compound 48/80-induced mortality associated with anaphylaxis was studied in rats. PTFE inhibited compound 48/80-induced anaphylaxis 100% with a dose of 1.6 mg/g body weight (BW) 1 h before or 5 min after injection of compound 48/80. PTFE inhibited compound 48/80-induced anaphylaxis almost 100% with doses above 0.4 mg/g BW intraperitoneally administered. PTFE (1-1000 micrograms/ml) also dose-dependently inhibited the histamine release induced by compound 48/80 (5 micrograms/ml) in rat peritoneal mast cells. The level of cAMP in peritoneal mast cells, when PTFE was added, increased transiently, and significantly increased 53-fold at 10 s compared with that of basal cells. Moreover, PTFE inhibited intracellular calcium release induced by compound 48/80. These results suggest that PTFE has antianaphylactic activity by stabilizing the peritoneal mast cell membrane.

Inhibition of immediate-type allergic reaction by Rosa davurica Pall. in a murine model.

We studied the effect of Rosa davurica Pall. (Rosaceae) fruits (RdF) on immediate-type allergic reactions. RdF completely inhibited compound 48/80-induced systemic anaphylactic shock at the dose of 1 g/kg. When RdF was given as pretreatment, at concentrations ranging from 0.0001 to 1 g/kg, the serum histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. RdF inhibited the passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) IgE antibody dose dependently. RdF also inhibited the histamine release induced by compound 48/80 or anti-DNP IgE from the rat peritoneal mast cells (RPMC). Moreover, RdF had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results indicate that RdF may contain compounds with actions that inhibit mast cell degranulation in the rat.

Effect of Polygala tenuifolia root extract on the tumor necrosis factor-alpha secretion from mouse astrocytes.

We investigated whether an aqueous extract of Polygala tenuifolia root (PTAE) inhibits secretion of tumor necrosis factor-alpha (TNF-alpha) from primary cultures of mouse astrocytes. PTAE dose-dependently inhibited the TNF-alpha secretion by astrocytes stimulated with substance P (SP) and lipopolysaccharide (LPS). Interleukin-1 (IL-1) has been shown to elevate TNF-alpha secretion from LPS-stimulated astrocytes while having no effect on astrocytes in the absence of LPS. We therefore also investigated whether IL-1 mediated inhibition of TNF-alpha secretion from primary astrocytes by PTAE. Treatment of PTAE to astrocytes stimulated with both LPS and SP decreased IL-1 secretion to the level observed with LPS alone. Moreover, incubation of astrocytes with IL-1 antibody abolished the synergistic co-operative effect of LPS and SP. These results suggest that PTAE may inhibit TNF-alpha secretion by inhibiting IL-1 secretion and that PTAE has an anti-inflammatory activity on the central nervous system curing some pathological disease states.

Effect of Syzygium aromaticum extract on immediate hypersensitivity in rats.

We investigated the effect of aqueous extract of Syzygium aromaticum (L.) Merr. et Perry (Myrtaceae) flower bud (SAFB) on immediate hypersensitivity. SAFB inhibited compound 48/80-induced systemic anaphylaxis in rats (IC50 = 31.25 mg/kg, i.p.). SAFB also inhibited local immunoglobulin E (IgE)-mediated passive cutaneous anaphylactic reaction (IC50 = 17.78 mg/kg, i.v.; IC50 = 19.81 mg/kg, p.o.). When SAFB was pretreated at concentrations ranging from 25 to 1000 mg/kg, the serum histamine levels were reduced in a dose-dependent manner. Moreover, SAFB dose-dependently inhibited histamine release from rat peritoneal mast cells (RPMC) by compound 48/80 or anti-dinitrophenyl IgE. When SAFB was added, the level of cAMP in RPMC transiently and significantly increased about 47-fold at 10 s compared with that of basal cells. These results indicate that SAFB inhibits immediate hypersensitivity by inhibition of histamine release from mast cells in vivo and in vitro.

Daeganghwal-tang inhibits the stem cell factor-induced migration and inflammatory cytokines secretion in mast cells.

Traditional Oriental medicinal prescription, Daeganghwal-tang (DGHT) has been used for the treatment of rheumatoid arthritis (RA) in Korea. However, its effect in experimental models remains unknown. Recent reports suggest that in patients with RA, synovial mast cells increase in number and show signs of activation and inflammatory cytokines secretion. Our results show that stem cell factor (SCF) is a potent chemotactic factor for the mast cells in vitro. The chemotactic response to SCF was blocked by DGHT. When DGHT (1mg/ml) was added, the secretion of tumor necrosis factor (TNF)-alpha, interleukin (IL)-1beta and IL-6 was inhibited by 60.1, 81.8, 72.5%, respectively in phorbol 12-myristate 13-acetate (PMA) plus calcium ionophore A23187-stimulated HMC-1 cells. In addition, the expression of TNF-alpha mRNA in HMC-1 cells was inhibited by DGHT (1mg/ml). These findings indicate that DGHT inhibits SCF-induced migration and PMA plus calcium ionophore-stimulated inflammatory cytokines secretion in mast cells.

Effect of Vitex rotundifolia on immediate-type allergic reaction.

We investigated the effect of aqueous extract of Vitex rotundifolia (L.) (Verbenaceae) fruits (VRFE) on the immediate-type allergic reactions in vivo and in vitro. VRFE (10(-4)-1.0 g/kg) dose-dependently inhibited systemic allergic reaction induced by compound 48/80. When VRFE was employed in a systemic allergic reaction test, the plasma histamine levels were reduced in a dose-dependent manner. VRFE (5x10(-1) and 1.0 g/kg) inhibited passive cutaneous anaphylaxis activated by anti-dinitrophenyl (DNP) IgE. VRFE (10(-3)-1.0 mg/ml) also dose-dependently inhibited the histamine release from the rat peritoneal mast cells (RPMC) by compound 48/80 or anti-DNP IgE. Moreover, VRFE (10(-3) mg/ml) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results suggest that VRFE may be beneficial in the regulation of immediate-type allergic reaction.

Inhibitory action of water soluble fraction of Terminalia chebula on systemic and local anaphylaxis.

We investigated the effects of the water soluble fraction of Terminalia chebula (Combretaceae) (WFTC) on systemic and local anaphylaxis. WFTC administered 1h before compound 48/80 injection inhibited compound 48/80-induced anaphylactic shock 100% with doses of 0.01-1.0 g/kg. When WFTC was administered 5 or 10 min after compound 48/80 injection, the mortality also decreased in a dose-dependent manner. Passive cutaneous anaphylaxis was inhibited by 63.5+/-7.8% by oral administration of WFTC (1.0 g/kg). When WFTC was pretreated at concentrations ranging from 0.005 to 1.0 g/kg, the serum histamine levels were reduced in a dose-dependent manner. WFTC (0.01-1.0 mg/ml) also significantly inhibited histamine release from rat peritoneal mast cells (RPMC) by compound 48/80. However, WFTC (1.0 mg/ml) had a significant increasing effect on anti-dinitrophenyl IgE-induced tumor necrosis factor-alpha production from RPMC. These results indicate that WFTC may possess a strong antianaphylactic action.

Inhibitory effect of mast cell-dependent anaphylaxis by Gleditsia sinensis.

We investigated the effect of aqueous extract of Gleditsia sinensis thorns (Leguminosae) (GSAE) on the mast cell-dependent anaphylaxis. GSAE (0.005 to 1 g/kg) dose-dependently inhibited systemic anaphylaxis induced by compound 48/80 in rats. GSAE (0.1 and 1 g/kg) also significantly inhibited local anaphylaxis activated by anti-DNP IgE. When GSAE was pretreated at the same concentrations with systemic anaphylaxis, the plasma histamine levels were reduced in a dose-dependent manner. GSAE (0.001 to 1 mg/ml) dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-DNP IgE. The level of cyclic AMP in RPMC, When CSAE (1 mg/ml) was added, transiently and significantly increased about fourfold compared with that of basal cells. Moreover, GSAE (0.01 and 0.1 mg/ml) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results suggest a possible use of GSAE in managing mast cell-dependent anaphylaxis.

Spirobenzylisoquinoline alkaloids from Corydalis ochotensis.

Separation of the alkaloids from the aerial parts of Corydalis ochotensis afforded a new spirobenzylisoquinoline alkaloid, 8-O-acetylcorysolidine along with two known spirobenzylisoquinoline alkaloids, isoochotensine and corysolidine.

Benzo[c]phenanthridine alkaloids from Corydalis incisa.

Six benzo[c]phenanthridine alkaloids, corynoline (1), acetylcorynoline (2), corynoloxine (3), luguine (4), 6-oxocorynoline (5), and 12-hydroxycorynoloxine (6) were isolated from the aerial parts of Corydalis incisa, and 6 was isolated for the first time from nature. The structure was elucidated by NMR techniques.

Suppression of anaphylactic reaction in murine bySiegesbeckia pubescens.

The aqueous extract ofSiegesbeckia pubescens (SPAE) inhibited compound 48/80-induced systemic anaphylaxis 100% with the dose of 1.0, 0.5 mg/g body weight (BW) at 1 h before or 5 min, 10 min after intraperitoneal injection of compound 48/80. The passive cutaneous anaphylaxis (PCA) reaction also inhibited to 78.5% by oral administration of SPAE (1.0 mg/g BW). When SPAE pretreated on mice at concentrations ranging from 0.0001 to 1.0 mg/g BW, the serum histamine levels were reduced in a dose-dependent manner. Moreover, SPAE (100-800 mug/ml) dose-dependently inhibited the histamine release from peritoneal mast cells (RPMC) by compound 48/80 (5 mug/ml). Analysis by microscopic appearance observation revealed that SPAE (500 mug/ml) stabilized the RPMC membrane. Therefore, these findings indicate that SPAE inhibits anaphylactic reactions through stabilization of mast cell membrane.

Antiallergic action of Magnolia officinalis on immediate hypersensitivity reaction.

We studied the effect of aqueous extract of Magnolia officinalis bark (Magnoliaceae) (MOAE) on the immediate hypersensitivity reaction. MOAE (0.01 to 1 g/kg) dose-dependently inhibited compound 48/80 induced systemic anaphylaxis in rats. MOAE (0.1 and 1 g/kg) also significantly inhibited local immunoglobulin E (IgE)-mediated passive cutaneous anaphylactic reaction. When MOAE was pretreated at concentrations ranging from 0.01 to 1 g/kg, the levels of plasma histamine were reduced in a dose-dependent manner. MOAE (0.001 to 1 mg/ml) dose-dependently inhibited the histamine release from rat peritoneal mast cells (RPMC) activated by compound 48/80 or anti-dinitrophenyl (DNP) IgE. The level of cyclic AMP (cAMP) in RPMC, when MOAE was added, significantly increased compared with that of the normal control. Moreover, MOAE (0.01 to 1 mg/ml) had a significant inhibitory effect on anti-DNP IgE-induced tumor necrosis factor-alpha production from RPMC. These results indicate that MOAE inhibits immediate hypersensitivity reaction in vivo and in vitro.

Anti-allergic components from the peels of Citrus unshiu.

In a bioassay-guided search for anti-allergic compounds from higher plants of Korea, polymethoxyflavones, 3',4',5,6,7,8-hexamethoxyflavone (1), 5-hydroxy-3',4',6,7,8-pentamethoxyflavone (II) and 3',4',5,7,8,-pentamethoxyflavone (III) have been isolated from the immature peels of Citrus unshiu. Structures of these compounds were elucidated on the basis of spectroscopic techniques. Compounds I and II inhibited dose-dependently histamine release from the rat peritoneal mast cells activated by compound 48/80 or anti-DNP IgE.

Effect of Alpinia oxyphylla fruit extract on compound 48/80-induced anaphylactic reactions.

The effect of the aqueous extract of Alpinia oxyphylla Miq. (Zingiberaceae) fruits (AOFE) on anaphylactic reaction was investigated. AOFE completely inhibited compound 48/80-induced systemic anaphylactic shock at dose of 1.0 g/kg. When AOFE was pretreated at concentrations ranging from 0.01 to 1.0 g/kg, the plasma histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. AOFE also inhibited the histamine release from rat peritoneal mast cells (RPMC) by compound 48/80. The level of cAMP in RPMC, when AOFE was added, transiently and significantly increased about 4-fold compared with that of basal cells. These results indicate that AOFE may be beneficial in the treatment of non-specific anaphylactic reactions.

Inhibition of mast cell-mediated anaphylaxis by sochungryong-tang.

According to traditional Asian philosophy, sochungryong-tang (S-Tang) is a prescription for treating exterior syndrome. In this study, we investigated the effect of S-Tang on mast cell-mediated anaphylaxis. S-Tang completely inhibited compound 48/80-induced systemic anaphylactic shock at a dose of 100 mg/kg. When S-Tang was given as pretreatment at concentrations ranging from 1 to 1000 mg/kg, the serum histamine levels induced by compound 48/80 were reduced in a dose-dependent manner. S-Tang inhibited the local anaphylaxis activated by anti-dinitrophenyl (DNP) IgE antibody, and also inhibited the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. These results indicate that S-Tang may contain substances with actions that inhibit mast cell degranulation.

A rare case of interparietal incisional hernia from 8 mm trocar site after robot-assisted laparoscopic prostatectomy.

Trocar site hernia arising from 8 mm robotic port is very rare despite the increasing prevalence of robot-assisted surgeries. To date, there had been only a single case reported in the literature. We report a case of small bowel obstruction secondary to an interparietal trocar site incisional hernia after robot-assisted laparoscopic prostatectomy. Meticulous closure of 8 mm robotic trocar sites associated with large peritoneal defect at the end of surgery should be performed.

Mosla punctulata Inhibits Mast Cell-mediated Allergic Reactions Through the Inhibition of Histamine Release and Inflammatory Cytokine Production.

Allergic inflammatory diseases such as food allergy, asthma, sinusitis and atopic dermatitis are increasing worldwide. This study examined the effects of aqueous extract of Mosla punctulata on mast cell-mediated allergic inflammation and studied the possible mechanism of action. Aqueous extract of Mosla punctulata inhibited compound 48/80-induced systemic and immunoglobulin E-mediated local anaphylaxis and it also reduced intracellular calcium level and down-streamed histamine release from mast cells. In addition, aqueous extract of Mosla punctulata decreased gene expression and secretion of tumour necrosis factor alpha, an important proinflammatory cytokine, in mast cells. The inhibitory effect on tumour necrosis factor alpha expression was nuclear factor kappa B dependent. The results indicate that aqueous extract of Mosla punctulata inhibited mast cell-mediated allergic inflammatory reaction by suppressing histamine release and expression of tumour necrosis factor alpha, and involvement of calcium and nuclear factor kappa B in these effects. Hence it can be concluded that, the aqueous extract of Mosla punctulata might be a possible therapeutic candidate for allergic inflammatory disorders.