木兰脂素抗炎、抗过敏作用的实验研究

目的：研究辛夷提取物－木兰脂素抗炎,抗过敏作用。方法：采用卵白蛋白（OA）引起的致敏豚鼠离体回肠实验和大鼠肥大细胞脱颗粒实验研究木兰脂素的抗过敏作用,醋酸致小鼠毛细血管通透性增加实验和角叉菜胶致小鼠足跖肿以及炎症组织中PGE2含量测定实验研究木兰脂素的抗炎作用。结果：木兰脂素能显著抑制致敏豚鼠离体回肠的过敏性收缩,明显阻止大鼠肥大细胞脱颗粒,对小鼠毛细血管通透性增加和小鼠足跖肿胀均有显著的抑制作用,并能降低炎症组织中PGE2的含量。结论：木兰脂素具有较强的抗过敏、抗炎作用。     

熟地黄和生地黄配伍的四物汤抗过敏作用的研究

目的：观察熟地（黄）和生地（黄）配伍的四物汤对实验性变态反应的影响，比较其作用差异。方法：以compound48／80诱导小鼠过敏性休克，皮肤搔抓反应及大鼠腹腔肥大细胞释放组胺，分别测定1h内的死亡率、10min内的搔抓次数及组胺的释放量和残留量（荧光法）；以氯化钴诱导小鼠耳廓迟发型超敏反应，测定耳廓肿胀度。结果：熟地配伍的四物汤（500mg/kg)显著抑制compound48／80所致小鼠过敏性休克，而生地配伍的四物汤无明显影响，熟地和生地配伍的四物汤（500mg/kg）均明显抑制compound48／80诱导的小鼠搔抓反应及氯化钴所致小鼠迟发型超敏反应，前者的作用强于后者；体外试验中，熟地或生地配伍的四物汤（100－400μg/ml）抑制compound48／80诱导的大鼠肥大细胞释放组胺，前者的作用强于后者，熟地提取物减少组胺释放，而生地提取物无明显作用。结论：熟地配伍的四物汤对肥大细胞依赖性速发型变态反应及T淋巴细胞依赖性迟发型变态反应的抑制作用均强于生地配伍的四物汤，熟地和生地作用和差异可能是其主要原因。     

炒紫苏子醇提物对肥大细胞脱颗粒及组胺释放的影响

目的肥大细胞脱颗粒及组胺释放是过敏性疾病发生机制中的重要环节之一,研究炒紫苏子醇提取物对其的作用,为研制和开发新的抗过敏中药提供实验依据。方法制备致敏大鼠腹腔肥大细胞悬液,体外和不同浓度药物作用,观察脱颗粒百分率和组胺释放水平,判断药物的抗过敏作用。除正常组(非致敏组)外,设空白对照组、炒紫苏子醇提取物320、640、1280μg/mL各剂量组、色甘酸钠阳性对照组、木犀草素组共7组。并对实验各个因素进行优化。结果经过最佳实验条件的优化,选择药物与肥大细胞作用时间为20min,OVA的触发浓度为20μg/mL。炒紫苏子醇提取物320、640、1280μg/mL各剂量组和木犀草素组均能明显降低IgE所致的I型过敏反应肥大细胞脱颗粒百分率(P<0.01),也明显降低其组胺释放(分别为P>0.05,P<0.01,P<0.01,P<0.01),并表现明显的剂量依赖关系。结论炒紫苏子醇提取物具有明显的肥大细胞脱颗粒及组胺释放等抗过敏作用。     

黄芩苷局部皮肤给药对小鼠血管通透性及豚鼠离体回肠收缩的影响

黄芩为唇形科植物黄芩Scutellaria baicalensis Georgi的干燥根，主要成分为黄酮类化合物，黄芩对甲基一甲氧基一苯乙胺和甲醛的聚合物所致敏的大鼠腹腔肥大细胞释放组胺及促使抗原与免疫球蛋白(IgE)的反应均有抑制作用。黄芩苷局部皮肤给药对小鼠血管通透性作用的研究，国内外均未见报道。本实验探讨黄芩苷局部皮肤给药对磷酸组胺引起的小鼠皮肤毛细血管通透性增高及豚鼠离体回肠收缩的影响。     

锦灯笼配方颗粒中木犀草素的HPLC法含量测定

目的：对中药锦灯笼配方颗粒中木犀草素进行含量测定。方法：采用Agilent1100高效液相色谱仪，EclipseXDB—C18色谱柱，流动相为甲醇：水（50：50），流速为1．0mL／min。结果：木犀草素在0．0143—0．0715μg线性关系良好（r=0．9996），锦灯笼配方颗粒中木犀草素含量平均值为0．14mg／g。结论：该方法简便易行、实验数据准确可靠，对于锦灯笼配方颗粒的内在质量评价有了提高。     

Effects of point application on celiac mast cell degranulation in mice with allergic rhinitis: An experimental study

目的：研究不同处理方法对卵白蛋白（Ovum Albumin,OVA）变应性鼻炎小鼠模型腹腔肥大细胞脱颗粒的影响。方法：将60只雌性小鼠随机分为5组,每组12只,分别按不同方法处理后分离各组小鼠的腹腔肥大细胞,中性红染色后计算腹腔肥大细胞脱颗粒率。结果：正常对照组、OVA变应性鼻炎组、穴位敷贴组、激素对照组、磷酸盐缓冲液（Phosphate Buffer Saline,PBS）阴性对照组小鼠腹腔肥大细胞脱颗粒率分别为（15±6）％、（53±11）％、（37±13）％、（31±15）％、（47±14）％。OVA变应性鼻炎小鼠的腹腔肥大细胞有明显的脱颗粒现象：与OVA变应性鼻炎小鼠相比,穴位敷贴组及激素对照组小鼠的腹腔肥大细胞脱颗粒现象显著减轻;PBS对照组小鼠的腹腔肥大细胞脱颗粒现象没有明显变化。结论：推测穴位敷贴抗过敏机制为稳定肥大细胞膜,抑制肥大细胞脱颗粒,减少致炎介质产生。     

MTT法测定双黄连粉针中有效成分对大鼠腹腔肥大细胞的IC50

目的：采用MTT法测定双黄连粉针中三种组成药材提取物及三种主要有效成分对大鼠腹腔肥大细胞IC50。方法：将空白大鼠腹腔肥大细胞无菌条件下分离纯化后,高、中、低三个浓度与腹腔肥大细胞及一定量的MTT共同孵育后,测定490nm处OD值。结果：双黄连中三种药材提取物及主要有效成分对大鼠肥大细胞的IC5 0分别为：0.18mg/mL、12.0μg/mL、0.06mg/mL;0.5mg/mL、8.0μg/mL、0.04mg/mL。结论：通过对上述各组药物对大鼠腹腔肥大细胞的IC50的测定,为考察双黄连粉针中各物质对肥大细胞的脱颗粒作用奠定实验基础。     

凉血清营颗粒对Ⅰ型变态反应的影响

目的:研究凉血清营颗粒对Ⅰ型变态反应的影响.方法:用天花粉蛋白所致小鼠速发型超敏反应及大鼠同种异体被动超敏反应(PCA)观察对Ⅰ型变态反应的影响;用化合物48/80所致小鼠过敏性休克死亡及大鼠颅骨肥大细胞模型观察对肥大细胞脱颗粒的影响;用组织胺所致小鼠皮肤毛细血管通透性亢进试验观察对组织胺的直接对抗作用.结果:凉血清营颗粒能一定程度拮抗天花粉蛋白所致小鼠过敏性休克及大鼠PCA,并能明显抑制化合物48/80及天花粉蛋白所致腹腔或骨膜肥大细胞脱颗粒,还能明显对抗组织胺所致毛细血管通透性亢进.结论:凉血清营颗粒具有显著的抗Ⅰ型变态反应作用,此作用可能是其能抑制IgE生成、肥大细胞脱颗粒及直接拮抗组织胺作用的综合效果.     

麻黄连翘赤小豆汤血清对肥大细胞脱颗粒、组胺生成的影响

本用含药血清体外作用于大鼠腹腔肥大细胞引起脱颗粒及组织胺释放为指标，观察了麻黄连翘赤小豆汤对Ⅰ型变态反应的影响，结果含该方大鼠血清可明显减少肥大细胞脱颗粒，减少组织胺释放。     

蛇床子活性成分R2止痒作用及其机制研究

目的观察蛇床子活性成分R2的止痒作用，并探讨其作用机制。方法观察不同浓度的R2软膏对磷酸组织胺致痒，4-氨基吡啶（4-AP）致小鼠皮肤瘙痒，二甲苯致小鼠耳廓肿胀，小鼠同种被动皮肤过敏反应及2，4-二硝基氯苯（DNCB）诱发小鼠迟发超敏反应的影响。通过测定小鼠注射4-AP后皮肤与血中组胺的含量，并利用致敏大鼠腹腔肥大细胞脱颗粒，探讨其止痒作用机制。结果Rz可显著提高磷酸组织胺对豚鼠的致痒阈，抑制由4-AP所引起的小鼠皮肤瘙痒反应，对抗二甲苯所致的小鼠耳廓肿胀，抑制小鼠同种被动皮肤过敏反应及抑制由DNCB所诱发的小鼠迟发超敏反应。研究表明№具有抗组胺和抑制肥大细胞脱颗粒的作用。结论R2具有止痒、抗炎、抑制Ⅰ、Ⅳ型变态反应作用，其机制与抗组胺、稳定肥大细胞膜及抑制肥大细胞脱颗粒等有关。     

Effects of Lecythis pisonis Camb. (Lecythidaceae) in a mouse model of pruritus

Ethnopharmacological relevance

Lecythis pisonis Camb. (Lecythidaceae), is popularly known as “Sapucaia”. In traditional medicine, leaves are used for the treatment of pruritus.

Aim of the study

The present study is aimed to investigate the antipruritic effect of the ethanol extract from leaves of Lecythis pisonis (LPEE), fractions (hexane-LPHF, ether-LPEF and ethyl acetate-LPEAF) and mixture of triterpenes [ursolic and oleanolic acids (MT)] in mice and rats.

Materials and methods

The LPEE, LPHF, LPEF, LPEAF and MT were evaluated on scratching behavior induced by compound 48/80 in mice. In addition, LPEE, LPEF and MT were investigated on rat peritoneal mast cells degranulation induced by compound 48/80 (ex vivo study). The anti-inflammatory activity of LPEE and LPEF was investigated in rats using carrageenan-induced hind paw edema model. In the evaluation of the spontaneous motor activity, the LPEE was studied for its effect on spontaneous motor activity in an open-field test in mice.

Results

The scratching behavior induced by compound 48/80 was significantly inhibited in mice pretreated with LPEE, LPHF, LPEF, LPEAF and MT. The suppressive effect of LPEE, LPEF and MT was only partially antagonized by naloxone. In addition, the compound 48/80-elicited degranulation of rat peritoneal mast cells was also markedly reduced in animals pretreated with LPEE, LPEF and MT. In the anti-inflammatory test, LPEE decreased the paw edema at the third hour after carrageenan (Carr) administration. Moreover, LPEF also was able to inhibit the oedematogenic response evoked by carr at all analysed time points. In the open-field test, LPEE-pretreated mice showed no impairment of spontaneous locomotion. Furthermore, the LPEE demonstrated no overt toxicity up to an oral dose of 2 g/kg in an acute toxicity assay.

Conclusions

These results clearly indicate the antipruritic effects of Lecythis pisonis leaves and suggest that this effect may be related to a stabilizing action on mast cell membrane. Furthermore, these data support the traditional use of this plant against cutaneous pruritus.     

Inhibitory activity of Chrysanthemi sibirici herba extract on RBL-2H3 mast cells and compound 48/80-induced anaphylaxis

The effects of extracts from various oriental medicinal herbs on mast cell-mediated allergic reaction were investigated. Among them, Chrysanthemi sibirici herba ethanol extract exerted the potent inhibitory activity on antigen-induced degranulation in RBL-2H3 mast cells. Chrysanthemi sibirici herba dose-dependently inhibited DNP-BSA or compound 48/80-induced degranulation in RBL-2H3 mast cells, with IC(50) values of approximately 49 microg/ml and 76 microg/ml, respectively. This extract strongly suppressed compound 48/80-induced systemic anaphylaxis by 48.7% at a dose of 300 mg/kg in mice. Chrysanthemi sibirici herba also inhibited the expression of TNF-alpha and the activation of the MAP kinase, ERK1/2, which is critical for the production of inflammatory cytokines in mast cells, as indicated by the suppression of activating phosphorylation of ERK1/2. These results lead us to conclude that Chrysanthemi sibirici herba may be used clinically to treat various allergic diseases.     

Effects of ryo-kan-kyomi-sin-ge-nin-to extract on degranulation of and histamine release from rat mast cells.

The effects of Ryo-kan-kyomi-sin-ge-nin-to (RKSG)) extract, a medicinal agent traditionally used in China and Japan for treatment of asthma, on the degranulation of and histamine release from rat mast cells were studied. At a concentration of 5 mg/ml RKSG, degranulation of mast cells stimulated either by antigen (DNP-Ascaris) or compound 48/80 was markedly suppressed. At a concentration of 1-5 mg/ml RKSG, histamine release from mast cells due to application of either antigen or compound 48/80 was inhibited in a dose-dependent fashion. These results suggest that RKSG may be useful for the treatment of type I allergy-related diseases.     

Shini-san inhibits mast cell-dependent immediate-type allergic reactions

Shini-San has been used for treatment of allergic disease in Korea. However, its effect in experimental models remains unknown. The mast cell plays a pivotal role in initiating allergic response by secreting intracytoplasmic granular mediators such as histamine. The present report describes an inhibitory effect of Shini-San on mast cell-mediated immediate-type allergic reactions. Topical application of compound 48/80 can induce an ear swelling response in normal (WBB6F1(-)+/+) mice but not in congenic mast cell-deficient WBB6F1-W/WV mice. Shini-San inhibited concentration-dependent mast cell-dependent ear swelling response induced by compound 48/80 in normal mice. Shini-San inhibited concentration-dependent passive cutaneous anaphylaxis induced by anti-dinitrophenyl (DNP) immunoglobulin E (IgE) in rats by topical application. Shini-San also inhibited in concentration-dependent fashion the histamine release from the rat peritoneal mast cells by compound 48/80 or anti-DNP IgE. Moreover, Shini-San had a significant inhibitory effect on compound 48/80-induced systemic anaphylactic reaction. These results indicate that Shini-San inhibits immediate type allergic reactions by inhibition of mast cell degranulation in vivo and in vitro.     

木犀草素对LPS诱导的RAW264.7细胞COX-2及mPGES-1表达的影响

目的:探讨木犀草素对脂多糖(LPS)诱导RAW264.7细胞COX-2及mPGES-1表达的影响。方法:酶免疫测定法(EIA)检测木犀草素对PGE2生成的影响,逆转录聚合酶链反应(RT-PCR)检测COX-2及mPGES-1 mRNA的表达。免疫印迹法(Western blotting)检测COX-2及mPGES-1蛋白的表达。结果:木犀草素抑制LPS诱导的RAW264.7细胞PGE2的生成,同时下调LPS诱导的RAW264.7细胞COX-2及mPGES-1 mRNA和蛋白的表达。结论:木犀草素可以抑制PGE合成途径中两个诱导酶COX-2和mPGES-1的表达。     

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.     

Antipruritic effect of the single oral administration of German chamomile flower extract and its combined effect with antiallergic agents in ddY mice

The single peroral administration of the ethyl acetate extract or essential oil of German chamomile (Matricaria recutita L.) showed remarkable antipruritic effects in the compound 48/80-induced itch-scratching test in ddY mice, if suitable vehicle was used. The ethyl acetate extract or essential oil of German chamomile dissolved in the vehicle of 10% ethanol, 10% Tween 80 and 80% physiological saline was orally administrated 2 h before pruritus provocation by compound 48/80 subcutaneous injection. The ethyl acetate extract or essential oil of German chamomile showed significant dose-dependent inhibition of the compound 48/80-induced scratching without affecting spontaneous motor activity. The antipruritic effects of antihistamine H1 antagonists, oxatomide (10 mg/kg) and fexofenadine (10 mg/kg), were only partial in this test. However, the antipruritic effects of these agents were remarkably enhanced by the combined administration of the ethyl acetate extract of German chamomile (300 mg/kg). Thus, the co-medication with the ethyl acetate extract, or essential oil of German chamomile and antihistamines might be effective for the pruritus which could not be perfectly resolved alone by conventional antihistamines.     

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。     

Mast cell protecting effects of shilajit and its constituents

The effects of shilajit and the combined effects of its main constituents, fulvic acids (FAs), 4′-methoxy-6-carbomethoxybiphenyl (MCB) and 3,8-dihydroxy-dibenzo-α-pyrone (DDP), were studied in relation to the degranulation and disruption of mast cells against noxious stimuli. Shilajit and different combinations of FAs, MCB and DDP provided statistically significant protection to antigen-induced degranulation of sensitized mast cells, markedly inhibited the antigen-induced spasm of sensitized guinea-pig ileum, and prevented mast cell disruption induced by compound 48/80. The findings are appraised in view of the clinical use of shilajit in the treatment of allergic disorders in Ayurvedic medicine.     

Effects of the Chinese herbal medicine "saiboku-to" on histamine release from and the degranulation of mouse peritoneal mast cells induced by compound 48/80

The herbal medicine "Saiboku-to" (ST) has been used for the treatment of bronchial asthma. ST was found to inhibit histamine release from and the degranulation of mouse peritoneal mast cells induced by compound 48/80. Its inhibitory effects were concentration-related and somewhat weaker than disodium cromoglycate, a known inhibitor of histamine release and mast cell degranulation. From these results, ST could appear to be useful in the treatment of Type I allergic reactions.