Magnolol and honokiol: inhibitors against mouse passive cutaneous anaphylaxis reaction and scratching behaviors

The antiallergic effects of magnolol and honokiol, isolated from the bark of Magnolia obovata (family Magnoliaceae), were investigated both in vitro and in vivo. Magnolol and honokiol potently inhibited passive cutaneous anaphylaxis reactions in mice induced by IgE-antigen complex as well as compound 48/80-induced scratching behaviors. These constituents exhibited not only potent inhibitory activity on the degranulation of RBL-2H3 cells induced by IgE-antigen complex, with IC(50) values of 45 and 55 muM, respectively, but also inhibited the protein expressions of IL-4 and TNF-alpha. Based on these findings, magnolol and honokiol may improve IgE-induced allergic diseases.     

杨梅素对小鼠被动皮肤过敏反应的影响

目的研究杨梅素的抗过敏作用。方法采用小鼠同种、异种被动皮肤过敏反应(PCA)、右旋糖酐致小鼠全身瘙痒反应和2,4-二硝基氯苯(DNCB)所致小鼠耳廓皮肤迟发型超敏反应(DTH)实验,探讨杨梅素的抗过敏作用。结果杨梅素显著抑制小鼠同种、异种被动皮肤过敏反应和右旋糖酐引起的小鼠瘙痒反应,并能抑制DNCB诱导的DTH。结论杨梅素具有抗过敏作用,机制可能与抑制Ⅰ、Ⅳ型变态反应有关。     

Hinokiresinol inhibits IgE-induced mouse passive cutaneous anaphylaxis reaction

The antiallergic effect of hinokiresinol isolated from the whole plant of TRAPA Pseudoincisa S. at. Z. was measured in vitro and in vivo. Hinokiresinol not only potently inhibited beta-hexosaminidase release from RBL-2H3 cells induced by IgE, with an IC50 value of 98 microM, but also inhibited the proinflammatory cytokines IL-6, IL-4 and TNF-alpha in RBL-2H3 cells stimulated by IgE. Orally and intraperitoneally administered hinokiresinol potently inhibited the passive anaphylaxis reaction in mice induced by IgE.     

Local effect of single stranded polyadenylic acid on passive cutaneous anaphylaxis in mice.

Single stranded polyadenylic acid (Poly A) administered locally inhibited passive cutaneous anaphylaxis in mice. In experiments performed by equilibrium dialysis Poly A was able to bind histamine. The association constant of the reaction was determined, Ka = 1.3 +/- 0.2 x 10(5) I/M. One Poly A molecule can bind maximally two molecules of histamine dichloride. Poly A inhibited the antigen-induced release of histamine from the peritoneal rat mast cells when it was given together with sensitizing antibodies.     

Inhibitory effects of an ellagic acid glucoside, okicamelliaside, on antigen-mediated degranulation in rat basophilic leukemia RBL-2H3 cells and passive cutaneous anaphylaxis reaction in mice

Degranulation inhibitors in plants are widely used for prevention and treatment of immediate-type allergy. We previously isolated a new ellagic acid glucoside, okicamelliaside (OCS), from Camellia japonica leaves for use as a potent degranulation inhibitor. Crude extracts from leaves also suppressed allergic conjunctivitis in rats. In this study, we evaluated the in vivo effect of OCS using a pure sample and performed in vitro experiments to elucidate the mechanism underlying the extraordinary high potency of OCS and its aglycon. The IC(50) values for degranulation of rat basophilic leukemia cells (RBL-2H3) were 14 nM for OCS and 3 μM for aglycon, indicating that the two compounds were approximately 2 to 3 orders of magnitude more potent than the anti-allergic drugs ketotifen fumarate, DSCG, and tranilast (0.17, 3, and >0.3 mM, respectively). Antigen-induced calcium ion (Ca(2+)) elevation was significantly inhibited by OCS and aglycon at all concentrations tested (p<0.05). Upstream of the Ca(2+) elevation in the principle signaling pathway, phosphorylation of Syk (Tyr525/526) and PLCγ-1 (Tyr783 and Ser1248) were inhibited by OCS and aglycon. In DNA microarray-screening test, OCS inhibited expression of proinflammatory cytokines [interleukin (IL)-4 and IL-13], cytokine-producing signaling factors, and prostaglandin-endoperoxidase 2, indicating that OCS broadly inhibits allergic inflammation. During passive cutaneous anaphylaxis in mice, OCS significantly inhibited vascular hyperpermeability by two administration routes: a single intraperitoneal injection at 10 mg/kg and per os at 5 mg/kg for 7 days (p<0.05). These results suggest the potential for OCS to alleviate symptoms of immediate-type allergy.     

大鼠被动皮肤过敏试验影响因素探讨

目的：系统性研究大鼠被动皮肤过敏试验（ PCA）的影响因素,为选择敏感试验条件和提高试验预测的准确性提供依据。方法：采用卵白蛋白（ OVA）为PCA试验抗原,在不同条件下（ OVA致敏剂量、添加佐剂、佐剂用量、致敏和被动致敏动物体质量及致敏时间）进行PCA试验。结果：①在不加弗氏完全佐剂（FCA）的条件下,在2．5～80 mg／只剂量范围内均未得到 PCA 阳性结果。②FCA 显著增加大鼠肌体免疫应答的敏感性;加用 FCA时, OVA 用量宜选用5～20 mg／只。③以每只动物致敏1 ml计,0．4 ml FCA∶0．6 ml溶媒、0．5 ml FCA∶0．5 ml溶媒及0．6 ml FCA∶0．4 ml溶媒三种配比方式均可用于PCA试验。④推荐最佳实验动物体质量为200～300 g。⑤致敏后12～14 d,PCA蓝斑直径可达峰值,最短致敏周期为8 d。结论：OVA致敏剂量、添加佐剂、佐剂用量、致敏和被动致敏动物体质量及致敏时间因素均对PCA结果有影响,试验时应予以重视。     

Deoxypodophyllotoxin, a naturally occurring lignan, inhibits the passive cutaneous anaphylaxis reaction

This study examined the effect of a podophyllotoxin derivative, deoxypodophyllotoxin (anthricin), which is a medicinal herb product isolated from Anthriscus sylvestris Hoffm. Deoxypodophyllotoxin was tested in a rat PCA (passive cutaneous anaphylaxis) assay by administering deoxypodophyllotoxin intraperitoneally (1.0 to 10 mg/kg, i.p.) and intravenously (0.25 to 1.0 mg/kg, i.v.). Deoxypodophyllotoxin dose-dependently inhibited the PCA reaction activated by anti-dinitrophenyl (DNP) IgE. The PCA inhibitory activity of deoxypodophyllotoxin was stronger than those of prednisolone and indomethacin, which were used as positive controls. These results suggest that deoxypodophyllotoxin may be beneficial in regulating the immediate-type allergic reaction.     

Topical application of luteolin inhibits scratching behavior associated with allergic cutaneous reaction in mice

The present study is aimed at evaluating the effects of luteolin on the scratching behavior associated with an allergic cutaneous reaction in mice. Elicitation of passive cutaneous anaphylaxis, and intradermal injections of compound 48/80, histamine or serotonin induced scratching behavior in ICR mice. Models of irritant contact dermatitis and allergic contact dermatitis were prepared by the topical application of 2,4-dinitrochlorobenzene (DNCB) on the ears of mice. Topical application of luteolin at concentrations of 20 and 100 mug/site significantly inhibited the number of scratching incidents associated with passive cutaneous anaphylaxis, and a similar tendency was also observed in histamine-, serotonin- and compound 48/80-evoked cutaneous reactions. The vascular permeability increase induced by passive cutaneous anaphylaxis or histamine injection was also significantly reduced by luteolin. Luteolin showed a potent inhibition on the ear thickness increase in models of irritant contact dermatitis and allergic contact dermatitis. In conclusion, luteolin significantly inhibited the scratching behavior associated with allergic cutaneous anaphylaxis. Its effects against pruritus are mainly attributed to its inhibition of mediator release from activated mast cells and direct antagonist effects on the released mediators which may act as local pruritogens.     

Inhibitory effects of glucocorticoids on increased vascular permeability caused by passive cutaneous anaphylaxis and some chemical mediators in rats

Effects of hydrocortisone, prednisolone and dexamethasone on IgE antibody-mediated homologous passive cutaneous anaphylaxis (PCA) and mediator-induced skin reactions were investigated. PCA and skin reactions were evoked at the same time in the dorsal skin of a rat. Administrations of glucocorticoids inhibited not only the PCA but also the skin reactions caused by histamine, serotonin and leukotriene (LT) C4 significantly. It is suggested, therefore, that glucocorticoids inhibit the increase of vascular permeability non-specifically. This action of glucocorticoids might contribute at least in part to its inhibitory effect on the PCA.     

被动皮肤过敏性评价中阳性对照的建立

目的：建立被动皮肤过敏性试验中准确且具可重复的阳性对照。方法：30只SD大鼠随机分为1个阴性对照(氯化钠注射液 弗氏不完全佐剂)和4个阳性处理组(阳性1组：卵白蛋白,阳性2组：卵白蛋白 百白破疫苗,阳性3组：卵白蛋白 弗氏不完全佐剂,阳性4组：卵白蛋白 弗氏完全佐剂及弗氏不完全佐剂),分别致敏5次制备抗血清,通过皮内注射被动致敏、48 h后激发,以皮内蓝斑有无及直径判定结果。结果：阴性对照和阳性1组、2组均未见蓝斑,阳性3和4组出现蓝斑,其1:2和1:8注射点蓝斑直径均大于5 mm。结论：被动皮肤过敏性试验中,致敏时结合使用弗氏佐剂可使阳性率达到100%,以保证试验的准确性和可重复性。     

Participation of histamine H1 and H2 receptors in passive cutaneous anaphylaxis-induced scratching behavior in ICR mice

Scratching behavior associated with passive cutaneous anaphylaxis was examined and compared to that induced by compound 48/80 or histamine in ICR mice. Elicitation of passive cutaneous anaphylaxis, and intradermal injections of compound 48/80, histamine or serotonin induced both scratching behavior and vascular permeability increase in ICR mice. In mast cell-deficient WBB6F1-W/Wv mice, although histamine induced scratching behavior and vascular permeability increase, passive cutaneous anaphylaxis was not observed. Cetirizine and terfenadine significantly inhibited the scratching behavior and vascular permeability increase caused by passive cutaneous anaphylaxis, compound 48/80 and histamine. The histamine H1 receptor antagonists inhibited the vascular permeability increase almost completely, whereas they failed to abolish the scratching behavior. Famotidine and ranitidine significantly inhibited the scratching behavior caused by histamine. The histamine H2 receptor antagonists did not affect the vascular permeability increase caused by histamine. The combination of cetirizine and ranitidine abolished the histamine-induced scratching behavior. The combination, however, failed to potentiate the inhibition of passive cutaneous anaphylaxis-induced scratching behavior significantly. The results indicated that histamine induces scratching behavior in ICR mice through both histamine H1 and H2 receptors, and that histamine plays a major role in passive cutaneous anaphylaxis-induced scratching behavior. Histamine might also play an important role in compound 48/80-induced scratching behavior.     

Inhibitory effects of a new antiallergic agent, azelastine, on passive cutaneous anaphylaxis and expulsion of Nippostrongylus brasiliensis

A new antiallergic agent, 4-(p-chlorobenzyl)-2-[N-methyl-perhydroazepinyl-(4)]-1-(2H)-phthalazinone hydrochloride (azelastine), was found to exert inhibitory effects on passive cutaneous anaphylaxis in the rat and on expulsion of Nippostrongylus brasiliensis from the rat intestine when this agent was administered intravenously at appropriate doses.     

Influence of lead and nickel on passive cutaneous anaphylaxis in the rat

Male rats, IOPS OFA strains, were injected s.c. with doses of lead acetate (0.5 to 30 mg Pb2+/kg/day) and nickel chloride (0.05 to 5 mg Ni2+/kg/day), for 3 days prior to being challenged for immediate hypersensitivity induced by ovalbumin/antiovalbumin system. The vascular permeability was statistically increased in animals of each group (P less than 0.001, Student's t-test). These results may indicate an action of lead and nickel salts on mast cells and their membranes, but also an action on the vasodilation induced by vasoactive amines.     

注射用替加环素的被动皮肤过敏性实验研究

目的:采用大鼠被动皮肤过敏试验(PCA)评价注射用替加环素的过敏性。方法:取8只雄性大鼠按体重分为4组制备抗血清;另取大鼠24只,雌雄各半,按以上分组进行致敏和激发试验。结果:被动皮肤过敏性试验顺利完成,未产生过敏反应。结论:大鼠被动皮肤过敏性试验显示注射替加环素未发生过敏反应。     

Biphasic response of cutaneous blood flow induced by passive cutaneous anaphylaxis in rats

In the immediate phase of passive cutaneous anaphylaxis, sensitized skin mast cells release various mediators when activated by antigen. The present study investigated the effects of the mediators on cutaneous blood flow at the antigen-antibody reaction site. Induction of passive cutaneous anaphylaxis produced a biphasic response consisting of an initial decrease, followed by a sustained increase, in the cutaneous blood flow. The initial phase was almost eliminated by the 5-hydroxytryptamine receptor antagonist methysergide, whereas the second phase was sensitive to the histamine H(2) receptor antagonist ranitidine. The histamine H(1) receptor antagonist chlorpheniramine, the denervation of sensory nerves with capsaicin, the cyclooxygenase inhibitor indomethacin, or the bradykinin B(2) receptor antagonist D-arginyl-L-arginyl-L-prolyl-trans-4-hydroxy-L-prolylglycyl-3-(2-thienyl)-L-alanyl-L-seryl-D-1,2,3,4-tetrahydro-3-isoquinolinecarbonyl-L-(2alpha,3beta,7abeta)-octahydro-1H-indole-2-carbonyl-L-arginine (HOE140) did not affect the blood-flow changes caused by the anaphylaxis. These results suggest that 5-hydroxytryptamine and histamine H(2) receptors mediate the initial decrease and the subsequent increase in cutaneous blood flow, respectively, induced by passive cutaneous anaphylaxis in rats.     

Inhibitory effect of chitosan-containing lotion on scratching response of hairless mice with atopic dermatitis-like dry skin

In this study, using a special diet-induced mouse model of atopic dermatitis, we tested the effect of chitosan-containing lotion (CL) on itch-related scratching associated with barrier-disrupted dry skin. HR-1 hairless mice fed a special diet exhibited apparent dry skin symptoms characterized by decreased skin hydration and increased transepidermal water loss. In the special diet-fed mice, scratching behavior was markedly enhanced for 60 min after oral administration of ethanol. When CL was applied once immediately after ethanol administration, the enhanced scratching response was significantly suppressed, but this effect was abolished within 30-40 min; when applied twice immediately and at 30 min, CL almost completely blocked scratching throughout 60 min. Comparison of CL and the chitosan-free vehicle showed that CL inhibited scratching more strongly and persistently than the vehicle, which transiently suppressed scratching only for 10 min after application. Although the decreased skin hydration was improved even by the vehicle, the increased transepidermal water loss was resolved only by CL. Skin surface temperature was much more reduced in CL-treated mice than in vehicle-treated mice. Collectively, CL has an antipruritic effect, which could be partly explained by recovery of skin barrier function and cooling of the skin.     

Immunopharmacological studies on TBX, a new antiallergic drug (1). Inhibitory effects on passive cutaneous anaphylaxis in rats and guinea pigs

The effects of 9-methyl-3-(1H-tetrazol-5-yl)-4H-pyrido[1,2-a]pyrimidin-4-one potassium salt (TBX), a new antiallergic drug, on passive cutaneous anaphylaxis (PCA) mediated by homologous IgE or IgG antibody were investigated in rats and guinea pigs. TBX (i.v. and p.o.) clearly inhibited IgE- and IgGa-mediated homologous PCAs in rats, without showing any inhibition of the skin reactions caused by histamine, serotonin and bradykinin in contrast to the inhibition of prostaglandin E1-induced skin reaction. Neither adrenalectomy nor propranolol treatment modified TBX's inhibition of the former PCA. With regard to tachyphylaxis to TBX, it was demonstrable in IgE-mediated homologous PCA in rats when they were pretreated with TBX (0.5 mg/kg, i.v.), followed 60 min later by a second dose of the drug (0.05 mg/kg, i.v.). There was no cross-tachyphylaxis between disodium cromoglycate (DSCG) and TBX. Homologous PCA caused by guinea pig IgE was also inhibited in a dose-dependent fashion by i.v. and p.o. administrations of TBX, although higher doses of TBX were needed to inhibit guinea pig PCA than the rat one. Interestingly, TBX showed more potent inhibition of both rat and guinea pig homologous PCAs than DSCG or tranilast. The results obtained indicate that TBX is an orally effective antiallergic agent displaying no antagonistic actions on the chemical mediators released.