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。     

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.     

穴位处胶原纤维在针刺大鼠"足三里"镇痛过程中的作用

目的：探讨针刺镇痛作用机制。方法：采用穴位注射Ⅰ型胶原酶法破坏穴区胶原组织结构。在急性佐剂关节炎大鼠模型上,以大鼠缩爪反射的潜伏期及肥大细胞脱颗粒率为观察指标,比较破坏与不破坏穴位区胶原组织结构2种情况下,针刺“足三里”捻转或提插手法对针刺镇痛效应及肥大细胞的影响。结果：破坏胶原结构后2种手法的镇痛效果明显削弱,且穴位肥大细胞脱颗粒也受到明显抑制。结论：穴区胶原纤维与肥大细胞在针刺镇痛效应的产生过程中起到了重要的作用,胶原纤维参与了针刺信号由穴位向靶器官传输与转换过程,而肥大细胞脱颗粒则始终与针刺镇痛效应呈正相关。     

手针刺激“足三里”穴局部肥大细胞活动与外周神经放电的相关性研究

目的:探讨穴位处肥大细胞的激活及穴区胶原与支配该区域的神经束放电变化之间的关系。方法:雄性SD大鼠30只,随机分为空白组、对照组、手针足三里组、手针+色甘酸钠组(穴区肥大细胞拮抗)、手针+胶原酶组(穴区胶原纤维溶解),每组6只。动物麻醉后,分离左侧"足三里"穴区坐骨神经干,手针提插刺激大鼠"足三里"穴20min,记录刺激前、刺激时和刺激后神经束诱发放电信号。对穴位组织进行组织学观察,统计肥大细胞脱颗粒率。结果:与对照组、手针+色甘酸钠组和手针+胶原酶组相比,手针足三里组诱发的坐骨神经放电活动明显增强(P<0.01);同时手针足三里组肥大细胞脱颗粒率显著升高(P<0.01),而手针+色甘酸钠组和手针+胶原酶组肥大细胞脱颗粒率与对照组相比差异无统计学意义(P>0.05)。结论:手针刺激大鼠"足三里"穴位可诱发支配该区域的外周神经束放电;穴位区注射色甘酸钠可抑制肥大细胞脱颗粒,针刺引起的外周神经放电减弱;穴位区注射胶原酶阻断针刺应力在穴位组织中传输,可以抑制肥大细胞脱颗粒,同样,针刺引起的外周神经放电也减弱。     

电针“足三里”穴对大白鼠腹膜腔肥大细胞的实验形态学观察

<正> 自提出经络现象与肥大细胞有关以来,不少学者观察到,电针刺激能引起穴位局部结缔组织中肥大细胞的脱粒反应。而肥大细胞在机体内分布极广,数量极多,并能产生多种生物活性物质。据报导,针刺“足三里”,循足阳明胃经能引起胃肠道蠕动的变化。设想电针刺激“足三里”对分布于胃肠系统腹膜腔的肥大细胞产生形态结构的影响。材料与方法实验选用体重200～300克的雄性大白鼠。实验分三组:甲组:电针刺激后肢“足     

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.     

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。     

激光针灸镇痛效应与穴区肥大细胞功能的关系

目的:观察低强度复合和单激光照射急性佐剂性关节炎大鼠"足三里"穴的镇痛效应及其与肥大细胞脱颗粒效应的关系.方法:采用清洁级SD大鼠66只,随机分为空白组、模型组、假照射组、10.6μm激光组、650 nm激光组和复合激光组,治疗时除空白、模型组外各组照射"足三里"穴30 min.除了空白组,其他5组通过左踝关节腔内注射完全弗式佐剂0.05 mL建立急性佐剂性关节炎模型.采用辐射热缩爪反射的方法,以大鼠缩爪反射的潜伏期为观察指标,比较各组的镇痛效果;通过穴位组织切片和甲苯胺蓝染色,离体对照激光照射前后穴位处局部肥大细胞脱颗粒率的变化.结果:激光照射后,650 nm激光组和复合激光组的痛阈显著高于模型组和假照射组(P＜0.01),650 nm激光组与复合激光组的肥大细胞脱颗粒率也显著高于模型组和假照射组(P＜0.001).10.6μm激光组的痛阈和肥大细胞脱颗粒率与模型组、假照射组比无显著差异.肥大细胞脱颗粒率与缩爪潜伏期的线性相关系数为0.737(P＜0.001).结论:低强度复合650 nm+10.6μm激光和单650 nm激光照射急性佐剂性关节炎大鼠"足三里"穴有显著镇痛效应;穴区中的肥大细胞脱颗粒率与镇痛效应呈正相关,在激光针灸镇痛效应的产生过程中起着重要作用.     

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。     

手针与电针“足三里”穴镇痛效应的穴位传入机制差异性分析

目的:观察手针与电针"足三里"穴位镇痛效应的穴位传入机制的差异性,分析这两种临床常用针刺方法的穴位始动信号的启动机制。方法:SD成年大鼠64只,随机分为正常组、模型组、手针组、电针组、胶原酶手针组、胶原酶电针组、色甘酸钠手针组和色甘酸钠电针组,每组8只。采用踝关节腔内注射完全弗氏佐剂(0.05 ml)建立大鼠佐剂性关节炎(AA)模型。在AA大鼠模型上,胶原酶手针和电针组于针刺前30 min在患侧"足三里"注射Ⅰ型胶原酶20μl(5 mg/ml)破坏局部组织胶原纤维,色甘酸钠手针和电针组于针刺前30 min在患侧"足三里"注射色甘酸钠20μl(0.02 g/ml)对局部肥大细胞进行功能屏蔽,并予以手针或电针。以大鼠缩爪反射潜伏期、痛阈变化率和肥大细胞脱颗粒率为镇痛效果和穴位激活的观察指标,比较穴区组织中胶原纤维对手针或电针"足三里"的镇痛效应及肥大细胞功能的影响。结果:Ⅰ型胶原酶或色甘酸钠"足三里"预处理后手针镇痛效果明显被削弱(P<0.05),而电针镇痛效果未受影响(P>0.05);两种预处理方法下,手针或电针穴位肥大细胞脱颗粒率均受到明显抑制(P<0.05)。结论:手针的针刺穴位始动信号可能是由胶原纤维参与介导的,并通过激活肥大细胞的功能,将有效信息经外周传入到中枢;而电针的针刺信号可能是直接激活外周神经感受器,由神经介导的。     

Anti-allergic activity of German chamomile (Matricaria recutita L.) in mast cell mediated allergy model

Ethnopharmacological relevance

Chamomile is most popular used medicinal plant and extensively consumed as a tea or tisanes. Traditionally this plant was used for treatment of many ailments such as allergy disorders and inflammatory mediated diseases.

Aim of the study

We investigated the effects of anti-allergic activity of Matricaria recutita L. on mast cell mediated allergic models.

Materials and methods

The protective effect of methanol extract of Matricaria recutita against compound 48/80 induced anaphylaxis and pruritis models for acute phase of hypersensitivity reactions were carried out. The late phase hypersensitivity reactions by compound 48/80 induced mast cell degranulation and histamine release from blood along with serum nitric oxide (NO), rat peritoneal fluid nitric oxide (NO) and bronchoalveolar fluid nitric oxide (NO) levels were measured.

Results

The methanol extract of Matricaria recutita L. showed inhibitory effects on anaphylaxis induced by compound 48/80 and significant dose dependent anti-pruritis property was observed by inhibiting the mast cell degranulation. Mast cell membrane stabilization activity was also observed in compound 48/80 induced mast cell activation. Dose dependent reduction in the histamine release, along with decreased release of serum, rat peritoneal and BAL fluid nitric oxide (NO) levels were observed.

Conclusion

These results suggest that the methanol extract of Matricaria recutita showed potent anti-allergic activity by inhibition of histamine release from mast cells.     

穴区肥大细胞功能与针刺缓解急性佐剂性关节炎大鼠疼痛效应的关系

目的:通过观察针刺镇痛效应与穴位区肥大细胞脱颗粒率的变化,研究针刺效应与穴位局部的肥大细胞功能之间的关系。方法:58只SD雄性大鼠随机分为正常组、模型组、留针组(仅留置针体在穴位处)、电针(EA,2/100Hz,0.5/1.0/1.5mA,“足三里”)组和手针(MA)组,在急性佐剂性关节炎(AA,完全弗氏佐剂0.05mL注射于动物踝关节)大鼠模型上,采用辐射热缩爪反射的方法,以大鼠缩爪反射的潜伏期为观察指标,比较电针、手针和留针组的镇痛效果。将穴位区的组织块制成石蜡切片,用甲苯胺蓝特殊染色法染色,观察其中肥大细胞变化的情况。结果:电针和手针对AA大鼠都有很明显的镇痛效应(与留针组比较P<0.01),而留针组无效(与模型组比较P>0.05),其中手针穴位处的效果更明显于电针。同时,电针和手针组的组织切片中肥大细胞脱颗粒率明显高于留针组(P<0.01)。结论:穴位组织中的肥大细胞在针刺镇痛效应的产生过程中起着重要作用,肥大细胞脱颗粒参与了针刺镇痛效应中针刺信号从穴位向靶器官传输与转换的过程。     

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.     

精制熊胆粉对IgE诱导的肥大细胞脱颗粒和血管通透性的影响

目的:观察精制熊胆粉的抗过敏作用,并探讨其可能的作用机制。方法:通过IgE诱导大鼠被动皮肤过敏反应(PCA)实验,用比色法测定熊胆粉在体内对肥大细胞影响。体外实验观察精制熊胆粉对IgE诱导组胺释放、细胞内钙摄入及肿瘤坏死因子-α、白细胞介素6以及NF-κB p65的影响。结果:体内实验显示,精制熊胆粉高(100mg/kg)、中(50mg/kg)剂量组能明显抑制大鼠PCA。体外实验显示,高(100μg/L)、中(50μg/L)浓度精制熊胆粉明显抑制了肥大细胞脱颗粒、组胺的释放、细胞内钙摄入以及肿瘤坏死因子-α、白细胞介素6以及NF-κB p65蛋白表达。结论:精制熊胆粉可能通过抑制NF-κB阻止肥大细胞的活化。     

The influence of pollen extracts (cernitin GBX and Cernitin T60) on allergic reactions

The influence of pollen extracts (Cernitin GBX and Cernitin T60) on some allergic reactions in animals and on the degranulation of mast cells and liberation of histamine has been studied. It was found that the cernitins possess antiallergic properties in vivo and in vitro. Both pollen preparations were able to inhibit the degranulation of mastocytes and histamine release from these cells. The results indicate that antiallergic properties of the cernitins might be dependent on their interaction with the mastocyte.     

Antiallergic effect of KOB03, a polyherbal medicine, on mast cell-mediated allergic responses in ovalbumin-induced allergic rhinitis mouse and human mast cells

Ethnopharmacological relevance

KOB03 is a polyherbal medicine consisting of five different herbs and has commonly been used for the treatment of various allergic diseases. However, its precise anti-allergic effect and mechanism remain unknown.

Aim of the study

The aim of this study was to investigate the effect of KOB03 on allergic responses through the regulation of mast-cell mediated allergic inflammation.

Materials and methods

To determine the effect of KOB03 on mast cell-mediated allergic reactions, we investigated the parameter changes of in vivo models such as compound 48/80-induced systemic anaphylaxis and ovalbumin (OVA)-induced allergic rhinitis, and the release of allergic inflammatory mediators such as histamine, immunoglobulin (Ig) E, and inflammatory cytokines via the MAPKs and NF-kappaB pathways.

Results

The oral administration of KOB03 at doses of 100 and 200 mg/kg inhibited histamine release and mortality in compound 48/80-induced anaphylactic rats. KOB03 also improved rhinitis symptoms, inhibited the histopathological changes of nasal mucosa, and decreased the serum levels of histamine, OVA-specific IgE and TNF-α in OVA-induced allergic rhinitis in mice. In vitro, KOB03 suppressed compound 48/80-induced histamine release by blocking mast cell degranulation. In addition, KOB03 inhibited the production of inflammatory cytokines such as TNF-α, IL-1β, IL-6 and IL-8 in PMA/A23187-stimulated HMC-1 mast cells by suppressing their gene expression and blocking the ERK1/2 and p38 MAPK and NF-κB pathways.

Conclusions

These results suggest that KOB03 has an anti-allergic effect by modulating mast cell-mediated allergic responses in allergic rhinitis.     

RBL-2H3和P815细胞用于建立体外肥大细胞脱颗粒模型的比较研究(英文)

目的:通过比较两种肥大细胞模型RBL-2H3和P815细胞激活后脱颗粒反应的差异,寻找早期、稳定、敏感的肥大细胞脱颗粒体外检测模型,为进一步建立基于细胞水平的过敏原和抗过敏药物的体外筛选模型提供一定的参考。方法:10μg.mL-1C48/80激活细胞15-60min,比色法检测组胺、β-氨基己糖苷酶及类胰蛋白酶等过敏介质的释放,中性红染色法进行形态学观察,流式细胞法检测AnnexinV阳性细胞率。结果:C48/80刺激P815、RBL-2H3细胞15-60min后组胺释放率均明显增加,并且相同刺激条件下,两种细胞模型的组胺释放率基本相当。C48/80刺激P815细胞15min以上,β-氨基己糖苷酶释放率、类胰蛋白酶释放率、Annexin V阳性细胞率和中性红染色脱颗粒细胞率均明显增加;但相同浓度的C48/80需刺激RBL-2H3细胞30-45min以上,上述指标才出现显著增加。在相同的刺激条件下,P815细胞过敏介质释放率、AnnexinV阳性率和脱颗粒率均高于RBL-2H3细胞。结论:同RBL-2H3相比,在相同刺激条件下,P815细胞活化后脱颗粒时间出现更早、程度更高。提示除RBL-2H3细胞,P815细...     

肥大细胞功能对针刺大鼠"足三里"镇痛效应的影响

目的:观察穴位肥大细胞与针刺效应之间的关系。方法:SD大鼠48只,随机分为正常对照组、穴位针刺组、旁开针刺组、色甘酸钠组、生理盐水组、色甘酸钠 针刺组、生理盐水 针刺组和色甘酸钠 对侧针刺组。采用大鼠尾部痛阈作为效应指标,在体观察针刺“足三里”提插捻转30 min过程中大鼠的甩尾潜伏期;并通过穴位组织切片染色,离体对照针刺前后穴位处局部肥大细胞脱颗粒率的变化,以及色甘酸钠注射对其的影响。结果:手针大鼠“足三里”穴具有显著镇痛作用,效果明显优于针刺旁开对照点;而在色甘酸钠屏蔽穴位肥大细胞的脱颗粒功能后,这种镇痛作用被明显地削弱。针刺后穴位处局部肥大细胞脱颗粒率显著提高;而注射色甘酸钠可以明显减少该脱颗粒现象。结论:穴位组织中的肥大细胞在针刺镇痛效应的产生过程中起着重要作用,肥大细胞脱颗粒参与了针刺镇痛效应的产生过程。     

Investigation of the Role of Mast Cells in Acupuncture Effects and Their Sensitivity to Physical Stimuli During TCM Treatment

Objective： To better understand the function of mast cells in acupuncture points （acupoints） in acupuncture-induced analgesia. The author tested their sensitivity to mechanical, thermo and light stimulation. Methods： The tail flick model was applied to measure analgesia in rats, and the author determined the density of mast cells in tissue slices and their degranulation ratio before/after acupuncture. The author also applied the patch-clamp technique to investigate activation of human mast cells （HMC1 cell line） by mechanical stress or noxious heat, and the author optically observed degranulation phenomena of mast cell in response to red laser light. Results： Manual stimulation by acupuncture at Zusanli （ST 36） of the rat resulted in analgesia and the effect was more pronounced than after stimulation of a sham point nearby the acupuncture point. A higher density of mast cells was found at Zusanli （ST 36） than at the sham point,     

Relationship Between and Laser Acupuncture Analgesia and the Function of Mast Cells in Adjuvant Arthritis Rats

Objective： To observe the analgesia effect and of low-level combined- and single-laser irradiation on Zusanli （ST 36） in rats and the relationship between mast cell degranulation and laser acupuncture analgesia. Methods： A total of 66 Sprague Dawley （SD） rats were randomly assigned into normal control, model control, sham irradiation, 10.6 μm laser （220 mW, 40 Hz） 650nm laser（36mW, continuous）, combined laser （10.6 μ m＋ 650 nm） groups. Arthritis mode 1 was established by injection of Complete Freund＇s Adjuvant （0.05 mL） into the left ankle joint. The paw withdrawal latency （PWL） was detected with a radiant heat algesimeter. Zusanli （ST 36） was irradiated with sham, single or combined laser for 30 min. After sacrifice under anesthesia （1% embutal）, tissues of Zusanli （ST 36） area were sampled, sliced （5 μ m） and stained with Toluidine Blue for skin for observing the mast cell degranulation. Results： Compared with model control and sham group, the pain threshold increased significantly in 650nm and combined laser groups （P〈0.01）, while remained no significant difference in 10.6/a m group. Compared with model and sham group, the degranulation ratios of mast cells in 650nm and combined laser group were significantly higher （P〈0.001）, while remained no significant difference in 10.6 ~ m group. The linear correlation coefficient between degranulation ratios of mast cells and PWL after laser acupuncture is 0.737 （P〈0.001）.Conclusion： Both 650 nm and combined laser stimulation of Zusanli （ST 36） can effectively raise pain threshold, and enhance degranulation ratio of mast cells at the stimulated acupoint. The result also suggested a linear correlation between degranulation ratio of mast cell and analgesia effect.