General immunological properties of fatsoluble (cernitin GBX) and water-soluble (cernitin T60) pollen extracts

The immunological properties of fat-soluble (Cernitin GBX) and water-soluble (Cernitin T60) pollen extracts were examined in vivo and in vitro. For investigations in vitro we used the water-soluble pollen extract (T60), and in vivo the fat-soluble form (GBX). The aim of the in vivo experiments was to evaluate their effect on IgG antibody production, their capability of rosette formation (E-RFC) and cell indicating IgM-plaqueforming cells (PFC) hemolysins. Also observations were made on the effect of the pollen extract on graft vs host reaction, the transplantation barrier and delayed hypersensitivity in relation to sheep red blood cells (SRBC). Its assumed effect on phagocytosis and blastic transformation was assessed in vitro. In both in vivo and in vitro systems the influence of T60 and GBX preparations on the population of T- and B-cells was tested. A relationship was revealed between immunological activity and the evaluated Cernitins. Both the Cernitins examined demonstrated moderate immunoregulatory properties, but the immunosuppressive component was predominant.     

舒宁康水提液抗过敏药效研究

目的:观察舒宁康水提液对肥大细胞脱颗粒及组胺释放的影响,研究其抗过敏作用.方法:以卵蛋白(OVA)为致敏原,在OVA质量浓度为40 mg·L-1作用30 min条件下,观察舒宁康0.1,1,10 g·L-1在体外对致敏大鼠肥大细胞脱颗粒及其释放组胺量的影响.结果:以色甘酸钠为阳性对照,舒宁康水提液具有明显的抑制肥大细胞脱颗粒作用,模型组、舒宁康水提液高、中、低剂量组及阳性对照组脱颗粒率分别为(47.53±5.57)％,(25.11±2.56)％,(35.71±6.99)％,(47.11±4.26)％,(22.87±6.52)％;舒宁康水提液对组胺的释放亦有显著的抑制作用,与模型组比,舒宁康水提液高、中、低组及阳性对照组的抑制率分别为16.85％,20.06％,6.58％,16.85％.结论:舒宁康水提液有良好的抗过敏作用,为将其开发成抗敏中药提供了实验依据.     

Evaluation of antiallergic and anti-anaphylactic activity of ethanolic extract of Sanseveiria trifasciata leaves (EEST) in rodents

Aim of the study

The leaves and rhizomes of Sansevieria trifasciata are used in folk medicine for treating bronchitis, asthma, cough, snake bite and insect bite etc. The ethanolic extract elicited analgesic, anti-inflammatory and antipyretic activity. Hence, it was decided to study the antiallergic activity of ethanolic extract of S. trifasciata (EEST) on various animal models as well as in vitro conditions, and also to understand possible mechanism of action.

Materials and methods

Ethanolic extract of S. trifasciata leaves (EEST) were prepared by cold maceration followed by concentration and evaporation under reduced pressure on a rotary evaporator to obtain semisolid mass. The various phytoconstituents were analyzed. The acute toxicity study of EEST was carried out in mice. The antiallergic and anaphylactic activities were evaluated using animal models viz. milk induced eosinophilia and leukocytosis, compound 48/80 induced mast cell degranulation, active and passive cutaneous anaphylaxis and histamine induced pedal edema. In addition, EEST effect on Shultz–Dale reaction in sensitized guinea pig ileum in ex vivo and antioxidant activity by free radical scavenging by DPPH method (in vitro) were also studied.

Results

EEST treatment at 100 mg/kg and 200 mg/kg p.o inhibited (a) milk-induced increased eosinophilia, leukocytosis, monocytes and neutrophils. (b) Prevented passive cutaneous and active anaphylactoid reactions. (c) Prevented compound 48/80 induced degranulation of sensitized mesenteric mast cells. (d) Inhibited histamine induced pedal edema formation significantly. EEST pretreatment inhibited Shultz–Dale reaction in guinea pig ileum and also elicited potent antioxidant activity.

Conclusion

Experimental findings demonstrate promising antiallergic and anti-anaphylactic activity of EEST and also elicited potent antioxidant activity. The antiallergic and anti-anphylactic activity might be due to inhibition of release of chemical mediators from mast cells largely by phytoconstituents like steroidal saponins, triterpenoids and flavonoids present in EEST.     

检测原代肥大细胞组胺释放分析中药注射剂致类过敏性

目的: 通过检测组胺释放考察19种临床常用中药注射剂致大鼠腹腔原代肥大细胞脱颗粒情况,为评价中药注射剂致类过敏性提供参考。 方法: 应用荧光法检测组胺,并进行条件优化以适用于中药注射剂所致肥大细胞释放组胺的检测。分离大鼠腹腔肥大细胞,分别将19种中药注射剂、阳性药C48/80和肥大细胞混合孵育1h,采用优化的荧光法检测组胺,再计算组胺释放率并进行统计学分析。 结果: 荧光法检测组胺最佳缩合反应时间为3 min,终止反应后7.5~30 min缩合物稳定性好;采用间接检测组胺释放率与直接检测组胺释放率两方法结果间无显著差异。痰热清、清开灵、参麦、丹参、血塞通、参附、双黄连、生脉、香丹、热毒宁、复方当归等11种中药注射剂致肥大细胞释放组胺作用明显。脉络宁、黄芪、艾迪、醒脑静、黄芪多糖、红花、盐酸川芎嗪、参芪等8种注射剂未见明显致肥大细胞释放组胺作用。 结论: 中药注射剂引发的过敏样反应可能大多为类过敏,检测组胺释放分析原代肥大细胞脱颗粒试验可以为评价中药注射剂的致类过敏性提供参考。     

仙蓟化斑胶囊对肥大细胞脱颗粒及组胺释放影响的研究

[目的]研究仙蓟化斑胶囊抗变态反应的作用机制,为后期实验及其临床治疗过敏性紫癜提供依据。[方法]制备致敏的大鼠及抗卵蛋白血清,给予不同浓度药物,采用大鼠被动皮肤过敏反应(PCA)实验,测定大鼠皮肤蓝斑直径,同时采用大鼠颅骨骨膜肥大细胞脱颗粒实验、大鼠肥大细胞释放组胺实验,测定脱颗粒百分率和组胺释放水平。[结果]仙蓟化斑胶囊可使蓝斑直径变小、可降低肥大细胞脱颗粒百分率、降低其组胺释放,并呈明显的剂量依赖关系。[结论]仙蓟化斑胶囊有抗变态反应的作用,明显抑制PCA、肥大细胞脱颗粒及组胺的释放。     

Antitumour potential of pollen extract on lewis lung carcinoma implanted intraperitoneally in syngeneic mice

A defined pollen extract of selected plants has been reported to possess some pharmacological activities on chronic prostatis or benign prostatic hyperplasia. This paper describes the antitumour potential of the water soluble fraction (Cernitin T60) of pollen extract against Lewis lung carcinoma implanted intraperitoneally in syngeneic mice. Cernitin T60 was not cytotoxic in cell cultures at concentrations up to 2.5 mg/mL, while it is significantly prolonged the life-span of mice carrying the tumour without any apparent side effects at 0.5 g/kg. In addition, Cernitin T60 demonstrated beneficial therapeutic effects in an additive fashion on the life-span of mice when it was combined with standard cytotoxic antitumour drugs such as adriamycin, cisplatin, vincristine, methotrexate, fluorouracil, or thioguanine. The antitumour potential of Cernitin T60 was completely abolished by treatment with inhibitors of macrophage functions (2-chloroadenosine or carrageenan), but not with a T-cell inhibitor (cyclosporin A). Cernitin T60 appears to be a potent immunostimulator of macrophages.     

防风醇提物对肥大细胞PAR-2及相关细胞因子的影响

目的:观察防风醇提物对肥大细胞过氧化物酶体增殖因子活化受体-2(PAR-2)及相关细胞因子的影响,探索防风抗过敏的新机制。方法:用胰蛋白酶刺激P815细胞的方法建立肥大细胞脱颗粒模型,设空白组,模型组,防风高、低剂量组(0.02,0.01 g·m L~(-1)),药物作用6 h后,采用ELISA检测细胞上清液中组胺,白细胞介素-4(IL-4),IL~(-1)3水平,Western blot,RTPCR检测PAR-2蛋白及其mRNA的表达。结果:与空白组比较,模型组组胺,IL-4,IL~(-1)3含量及PAR-2蛋白及mRNA的表达明显升高(P0.05,P0.01);与模型组比较,防风醇提物在体外抑制肥大细胞组胺,IL-4,IL~(-1)3含量及PAR-2蛋白及mRNA的表达(P0.05,P0.01)。结论:防风醇提物可能通过抑制PAR-2表达,阻断肥大细胞脱颗粒,且选择性减少相关细胞因子分泌,继而抑制肥大细胞"瀑布效应",达到抗过敏作用。     

水栀子及京尼平苷抑制RBL-2H3细胞脱颗粒作用的代谢组学研究

目的 基于细胞代谢组学的方法研究水栀子Gardenia jasminoside及其主要有效成分京尼平苷对RBL-2H3细胞脱颗粒模型的干预作用。方法 利用UPLC-QTOF-MS高分辨质谱检测RBL-2H3细胞的代谢轮廓改变,并应用主成分分析（PCA）和正交偏最小二乘判别分析（OPLS-DA）筛选出引起这种差异的代谢标志物;同时应用MEV软件对差异标志物水平的变化进行聚类分析和热图绘制。结果 分别得到与水栀子和京尼平苷作用相关的代谢标志物54和46个,其中共有标志物31个,且在2个给药组中变化趋势相同。由共有代谢标志物富集得到了受干扰的5条代谢通路：甘氨酸、天冬氨酸、谷氨酸代谢,谷胱甘肽代谢,组胺代谢,能量代谢,烟酰胺代谢。结论 水栀子和京尼平苷通过调节组胺代谢、氧化应激和能量代谢而抑制RBL-2H3细胞脱颗粒过程,且京尼平苷是水栀子发挥作用的主要药效物质基础之一。     

Pollen extracts reduce the hepatotoxicity of paracetamol in mice

Cernitins are preparations obtained from plant pollen which contain numerous compounds of potential biological significance. This work deals with the influence of cernitins upon acute paracetamol toxicity in mice. The survival rate and indices of hepatic injury: aminotransferase and alkaline phosphatase activities, bilirubin level in serum, glutathione and cytochrome P-450 content in liver, liver weight, histopathologic picture of the liver and presence of glycogen and lipids in stained liver sections, under different experimental protocols, were determined. It was found that cernitins are able to increase the survival rate of mice and reduce liver injury in acute paracetamol poisoning. Cernitins are more effective when administered after, rather than before, a dose of paracetamol. The possible mechanism through which cernitins may act is discussed.     

Lipid‐soluble components of honeybee‐collected pollen exert antiallergic effect by inhibiting IgE‐mediated mast cell activation in vivo

It was shown previously that bee‐collected pollen (bee pollen, BP), inhibited in vitro murine mast cell activation. This study further analysed the antiallergic effect of BP in vivo by measuring cutaneous mast cell activation using a passive cutaneous anaphylaxis reaction. Daily oral administration of BP to mice, dose‐dependently reduced the cutaneous mast cell activation elicited by IgE and specific antigens. Administration of BP also reduced the plasma concentration of malondialdehyde (MDA), an indicator of lipid peroxidation. The inhibitory effect of BP was mostly in a lipid‐ but not in water‐soluble fraction. The HPLC analysis of isoflavones in BP revealed that genistein was a major isoflavone. However, administration of genistein alone at the concentration found in BP, did not show an inhibitory effect as observed in whole BP, suggesting that component(s) other than genistein would be responsible for the inhibitory effect of BP. These results first reveal that lipid‐soluble components of BP exert an antiallergic action by inhibiting the FcåRI‐mediated cutaneous mast cell activation. Copyright © 2009 John Wiley & Sons, Ltd.     

5‐Lipoxygenase‐inhibitory constituents from Schizandra fructus and Magnolia flos

In order to establish the antiallergic properties of Schisandra fructus and Magnolia flos, several compounds isolated from these plants were tested for 5‐lipoxygenase (5‐LOX) inhibitory activity in vitro, for the first time. The compounds including schizandrins, schisandrols, gomisins, fargesin, eudesmin and lirioresinol B dimethyl ether, inhibited 5‐LOX‐catalysed leukotriene production from A23187‐treated rat basophilic leukemia (RBL‐1) cells at concentrations of 1–100 µm . In particular, constituents such as schisandrol A and gomisins showed potent inhibitory activity (IC₅₀s < 10 µm ) on 5‐LOX‐catalysed leukotriene production, but were much less active on cyclooxygenase‐2‐catalysed prostaglandin E₂ and inducible nitric oxide‐catalysed NO production. These compounds have the potential to be developed as novel antiallergic agents and may contribute to the antiallergic pharmacological use of these plant materials in Chinese medicine. Copyright © 2009 John Wiley & Sons, Ltd.     

Mast cell stabilization and antihistaminic potentials of Curculigo orchioides rhizomes

Aim of the study

To investigate the mast cell stabilization and antihistaminic activities of the rhizomes of Curculigo orchioides (COR). Extract of Curculigo orchioides Gaertn. (Fam. Amaryllidaceae) has been reported to possess immunostimulant, and anti-inflammatory potentials. In Indian traditional system of medicine it is also used as anti-asthmatic and anti-inflammatory.

Materials and methods

Estimation of histamine release is key parameter for evaluating any target for its anti-allergic potential. The stabilization potential of the alcoholic extract of COR (100–400 mg/kg) against mast cell degranulation was studied on isolated mice peritoneal mast cells. The antihistaminic activity was performed by determining the mortality rate of mice upon exposure to compound 48/80 and effect on inhibition of histamine release upon degranulation.

Results

The raised number of intact mast cells intimates that the COR stabilized the mast cell degranulation (60.96 ± 1.96%) and percentage antihistaminic potential of the extract (63.58 ± 1.8 inhibition at dose of 400 mg/kg) and it virtues further work towards the isolation of phytoconstituents from this plant.

Conclusion

This finding provides evidence that COR inhibits mast cell-derived immediate-type allergic reactions and mast cell degranulation.     

Antiallergic activity of a disaccharide isolated from Sanguisorba officinalis

The antiallergic activity of the natural disaccharide, 5-O-alpha-D-(3-C-hydroxymethyl)lyxofuranosyl-beta-D-(2-C-hydroxymethyl)arabinofuranose was evaluated using both in vivo and in vitro experimental models. Intravenously administered compound inhibited the passive cutaneous anaphylaxis response in rats in a dose-dependent manner (ED(50) = 9.6 mg/kg). The compound inhibited histamine release evoked by both compound 48/80 and calcium ionophore A23187 in rat peritoneal mast cells indicating that mast cell stabilization is the major mechanism of action for its antiallergic activity. In passively sensitized isolated guinea-pig hearts, an in vitro anaphylaxis model in which histamine release plays a key role for functional deterioration, the compound markedly diminished both coronary flow reduction and histamine release on challenge to the antigen. These data demonstrate that this antiallergic natural disaccharide exerts its effect via inhibition of mast cell mediator release.     

Mast cell stabilizing effect of glucan A and phosphatidyllycorine isolated from Crinum latifolium

The effect of glucan A and phosphatidyllycorine, isolated from Crinum latifolium L. (family Amaryllidaceae) was studied on the rate of degranulation of mast cells of albino rats. Different combinations of glucan A and phosphatidyllycorine (5–20 and 5–10 μg/mL, respectively) in vitro, produced statistically significant protection against Tween 80-induced degranulation, as well as to sensitized mast cells challenged with an antigen (horse serum). The combination (10–20 mg/kg), when administered in vivo, also provided protection against compound 48/80-induced degranulation of mast cells. The findings are appraised in view of the use of the total extract of C. latifolium, for the treatment of allergic disorders in Ayurvedic medicine.     

痛泻要方抑制致敏大鼠腹腔肥大细胞脱颗粒的研究

目的观察痛泻要方对化合物48/80(C48/80)诱导的大鼠腹腔肥大细胞脱颗粒的影响,探讨痛泻要方对肠易激综合征的作用机制。方法应用钙离子荧光指示剂Fluo-3,通过光学显微镜和激光共聚焦荧光显微镜,观察痛泻要方对致敏大鼠腹腔肥大细胞脱颗粒百分数及肥大细胞内游离钙浓度和组胺释放的影响。结果实验组(痛泻要方组)致敏大鼠腹腔肥大细胞脱颗粒百分数为13%,对照组为41%,2组有显著性差异(P<0.05);对照组和实验组致敏大鼠腹腔肥大细胞内游离钙浓度分别为(436.16±12.30)nmol/L和(78.31±3.28)nmol/L,2组相比有显著性差异(P<0.05);对照组和实验组大鼠腹腔肥大细胞上清液组胺浓度分别为(98.41±7.23)nmol/L和(33.23±21.50)nmol/L,2组相比有显著性差异(P<0.05)。结论痛泻要方对C48/80诱导的大鼠腹腔肥大细胞的脱颗粒过程有明显的抑制作用,表明痛泻要方是通过抑制肠道肥大细胞活化,从而减少其释放组胺等递质实现治疗作用的。     

Efficacy of a Nasal Spray from Citrus limon and Cydonia oblonga for the Treatment of Hay Fever Symptoms—A Randomized,Placebo Controlled Cross‐Over Study

Nasal spray from lemon and quince (LQNS) is used to treat hay fever symptoms and has been shown to inhibit histamine release from mast cells in vitro. Forty‐three patients with grass pollen allergy (GPA) were randomized to be treated either with placebo or LQNS for one week, respectively, in a cross‐over study. At baseline and after the respective treatments patients were provoked with grass pollen allergen. Outcome parameters were nasal flow measured with rhinomanometry (primary), a nasal symptom score, histamine in the nasal mucus and tolerability. In the per protocol population absolute inspiratory nasal flow 10 and 20 min after provocation was higher with LQNS compared to placebo (?37 ± 87 mL/s; p = 0.027 and ?44 ± 85 mL/s; p = 0.022). The nasal symptom score showed a trend (3.3 ± 1.8 in the placebo and 2.8 ± 1.5 in the LQNS group; p = 0.070) in favor of LQNS; the histamine concentration was not significantly different between the groups. Tolerability of both, LQNS and placebo, was rated as very good. LQNS seems to have an anti‐allergic effect in patients with GPA. Copyright © 2016 John Wiley & Sons, Ltd.     

Inhibitory effects of Cinnamomum cassia extract on atopic dermatitis-like skin lesions induced by mite antigen in NC/Nga mice

Aim of the study

: Cinnamomum cassia (C. cassia) has been traditionally used to treat allergic disease as well as dyspepsia, gastritis, and blood circulation disturbances. However, the antiallergic properties of C. cassia have not been fully verified using scientific tools. This study investigated the effectiveness of C. cassia extract (CCE) as an antiallergic agent in atopic dermatitis model and underlying mechanism.

Materials and methods

: The effect of CCE on mite antigen-treated NC/Nga mice was evaluated by examining skin symptom severity, levels of serum IgE, tumor necrosis factor-α (TNF-α), and histamine, skin histology, and mRNA expression of cytokines in the skin lesions. Moreover, the effect of CCE on TNF-α-and interferon-γ (IFN-γ)-induced chemokine production in human keratinocytes was investigated using ELISA.

Results

: CCE treatment of NC/Nga mice reduced the dermatitis score and the levels of serum IgE, histamine, and TNF-α. Histological examination showed inhibition of the thickening of the epidermis/dermis and reduced dermal infiltration of inflammatory cells. In skin lesions, mRNA expression of IL-4, TNF-α, and thymus and activation-regulated chemokine (TARC) was inhibited by CCE treatment. The production of TARC, macrophage-derived chemokine, and RANTES from IFN-γ-and TNF-α-stimulated human keratinocytes was suppressed by CCE treatment in a dose-dependent manner.

Conclusions

: CCE inhibits the development of atopic dermatitis-like skin lesions in NC/Nga mice by suppressing the T-helper 2 cell response.     

Pycnogenol® inhibits immunoglobulin E‐mediated allergic response in mast cells

IgE‐dependent mast cell activation is known to be associated with the allergic diseases. Pycnogenol^® (PYC) is a standardized extract of the bark of French maritime pine containing bioflavonoids with a potent antioxidant activity. The antiallergic activity of PYC was evaluated using both in vivo and in vitro experimental models. Oral administration of PYC significantly inhibited anti‐dinitrophenyl (DNP) IgE‐mediated passive cutaneous anaphylaxis in rats. In an in vitro study, PYC dose‐dependently reduced histamine release from rat peritoneal mast cells (RPMC) triggered by anti‐DNP IgE. PYC inhibited the protein expression and secretion of tumor necrosis factor‐α and interleukin‐6 in anti‐DNP IgE‐stimulated RPMC. Moreover, PYC decreased anti‐DNP IgE‐induced calcium uptake into RPMC. Furthermore, PYC suppressed nuclear factor‐kappa B activation. From these results, the clinical use of PYC in the mast cell‐mediated immediate‐type allergic diseases is proposed. Copyright © 2009 John Wiley & Sons, Ltd.     

Antiallergic and antihistaminic effect of two extracts of Capparis spinosa L. flowering buds

The antiallergic properties of two lyophilized extracts obtained from Capparis spinosa L. flowering buds (capers) by methanol extraction, carried out at room temperature (CAP-C) or with heating at 60 degrees C (CAP-H), were investigated.The protective effects of CAP-H and CAP-C, orally administered (14.28 mg[sol ]kg), were evaluated against Oleaceae antigen challenge-induced and histamine-induced bronchospasm in anaesthetized guinea-pigs. Furthermore, the histamine skin prick test was performed on humans, applying a gel formulation containing 2% CAP-C (the only extract able to protect against histamine-induced bronchospasm) on the skin for 1 h before histamine application and monitoring the erythema by reflectance spectrophotometry.The CAP-H showed a good protective effect against the bronchospasm induced by antigen challenge in sensitized guinea-pigs; conversely, a significant decrease in the responsiveness to histamine was seen only in CAP-C pretreated animals. Finally, the CAP-C gel formulation possessed a marked inhibitory effect (46.07%) against histamine-induced skin erythema.These two caper extracts displayed marked antiallergic effectiveness; however, the protective effect of CAP-H was very likely due to an indirect mechanism (for example, inhibition of mediator release from mast cells or production of arachidonic acid metabolites); conversely, CAP-C is endowed with direct antihistaminic properties. The different mechanisms of action of CAP-H and CAP-C may be related to a difference in the extraction procedure and, thus, in their qualitative[sol ]quantitative chemical profile.     

姜黄消痤搽剂抗过敏及止痒作用的实验研究

目的研究姜黄消痤搽剂的抗过敏、止痒作用。方法用卵白蛋白（OA）免疫豚鼠造成过敏模型，观察姜黄消痤搽剂对OA所致的豚鼠离体回肠平滑肌收缩的影响；在豚鼠腹腔注射OA致敏，观察姜黄消痤搽剂对OA引起的豚鼠腹腔肥大细胞脱颗粒的影响；以4氨基吡啶诱发小鼠舔体反应，观察姜黄消痤搽别对舔体反应的影响；以组胺使小鼠毛细血管通透性增加，观察姜黄消痤搽剂对通透性的影响从而观察姜黄消痤擦剂的抗过敏、止痒作用。结果姜黄消痤搽剂显著抑制OA所致的豚鼠离体回肠平滑肌过敏性收缩；显著抑制OA引起的肥大细胞脱颗粒；显著抑制4氨基吡啶诱发的小鼠舔体反应；显著抑制组胺引起的小鼠毛细血管通透性增加。结论姜黄消痤擦剂时多种动物模型有抗过敏作用，这种作用可能与其抑制肥大细胞脱颗粒和抗炎作用有关。