Mast cell tryptase and carboxypeptidase A expression in body fluid and gastrointestinal tract associated with drug-related fatal anaphylaxis

AIM: To investigate the expression of mast cell tryptase and carboxypeptidase A in drug-related fatal anaphylaxis.METHODS: The expression of mast cell tryptase and carboxypeptidase A in 15 autopsy cases of drugrelated fatal anaphylaxis and 20 normal autopsy cases were detected. First, the expression of mast cell tryptase was determined in stomach, jejunum, lung, heart, and larynx by immunofluorescence. Different tissues were removed and fixed in paraformaldehyde solution, then paraffin sections were prepared for immunofluorescence. Using specific mast cell tryptase and carboxypeptidase A antibodies, the expression of tryptase and carboxypeptidase A in gastroenterology tract and other tissues were observed using fluorescent microscopy. The postmortem serum and pericardial fluid were collected from drug-related fatal anaphylaxis and normal autopsy cases. The level of mast cell tryptase and carboxypeptidase A in postmortem serum and pericardial fluid were measured using fluor enzyme linked immunosorbent assay(FEIA) and enzyme linked immunosorbent assay(ELISA) assay. The expression of mast cell tryptase and carboxypeptidase A was analyzed in drug-related fatal anaphylaxis cases and compared to normal autopsy cases.RESULTS: The expression of carboxypeptidase A was less in the gastroenterology tract and other tissues from anaphylaxis-related death cadavers than normal controls. Immunofluorescence revealed that tryptase expression was significantly increased in multiple organs, especially the gastrointestinal tract, from anaphylaxis-related death cadavers compared to normal autopsy cases(46.67 ± 11.11 vs 4.88 ± 1.56 in stomach, 48.89 ± 11.02 vs 5.21 ± 1.34 in jejunum, 33.72 ± 5.76 vs 1.30 ± 1.02 in lung, 40.08 ± 7.56 vs 1.67 ± 1.03 in larynx, 7.11 ± 5.67 vs 1.10 ± 0.77 in heart, P < 0.05). Tryptase levels, as measured with FEIA, were significantly increased in both sera(43.50 ± 0.48 μg/L vs 5.40 ± 0.36 μg/L, P < 0.05) and pericardial fluid(28.64 ± 0.32 μg/L vs 4.60 ± 0.48 μg/L, P < 0.05) from the anaphylaxis group in comparison with the control group. As measured by ELISA, the concentration of carboxypeptidase A was also increased more than 2-fold in the anaphylaxis group compared to control(8.99 ± 3.91 ng/m L vs 3.25 ± 2.30 ng/m L in serum, 4.34 ± 2.41 ng/m L vs 1.43 ± 0.58 ng/m L in pericardial fluid, P < 0.05).CONCLUSION: Detection of both mast cell tryptase and carboxypeptidase A could improve the forensic identification of drug-related fatal anaphylaxis.

Treatment refractory mast cell leukemia with dominant gastrointestinal manifestation and concomitant skin symptoms:A case report

BACKGROUND Mast cell leukemia(MCL),a subtype of systemic mastocytosis(SM),is an extremely rare clinical entity characterized by a very poor prognosis.Chemotherapy,tyrosine kinase inhibitors,and allogeneic hematopoietic cell transplantation are the only treatment options,but they cannot provide the desired outcomes in most cases of MCL.However,other types of SM can be successfully treated.The disease has no specific manifestation,but gastroenterological symptoms are present in most cases.CASE SUMMARY The authors,hereby,report a case of a 46-year-old female patient diagnosed with MCL-the rarest subtype of SM.The patient presented to the gastroenterology clinic with multiple,various,and unspecific gastroenterological symptoms.Concomitance of skin lesions significantly contributed to a relatively prompt diagnosis.The serum tryptase level was extremely high and bone the marrow aspirate showed an infiltration of atypical mast cells.The disease was rapidly progressive and primary refractory to chemotherapy and the patient succumbed to the illness about a month after the initiation of treatment.CONCLUSION Despite its“hematological nature”,MCL,in most cases presents dominantly with unspecific gastroenterological symptoms.Thus,a high disease awareness among physicians other than hematologists is necessary to improve treatment outcomes.Serum tryptase level,due to its non-invasive nature and easy access,may serve as an initial step to estimate the probability of mastocytosis.

Mast cell activation syndrome:An up-to-date review of literature

Mast cells are a subtype of white blood cells and are involved in the immune system.These cells contain many chemical substances called mediators,which are involved in the allergic response.The fact that mast cells play a role in many events that require urgent intervention,especially anaphylaxis,has led to a more detailed study of these cells.The diseases also caused by dysfunctions of mast cells have been examined in many circumstances.For instance,mast cell activation syndrome is known as an augmented number of cells due to decreased cell death,resulting in clinical symptoms affecting many systems.The main common symptoms include flushing,hypotension,urticaria,angioedema,headache,vomiting and diarrhea.Although the underlying mechanism is not yet clearly known,we aim to review the literature in a broad perspective and bring together the existing knowledge in the light of the literature due to the diversity of its involvement in the body and the fact that it is a little known syndrome.

Effect of 1,25-dihydroxyvitamin D3 on mast cells tryptase in asthmatic guinea pigs

Objective:To explore the effect of 1,25-dihydroxyvitamin D3 on the mast cell tryptase(MCT) in asthmatic guinea pigs.Methods:A total of 60 male or female healthy guinea pigs were randomly divided into control group(group A),asthmatic group(group B).and 1,25-dihydroxyvitamin D3 group(group C),with 20 cases in each group.To establish asthmatic guinea pig models,1ml peanut oil was tilled into stomach in the morning in group A and group B.and 1 ml peanut oil with 1,25-dihydroxyvitamin D3 was filled into stomach in group C.Airway resistance(Re) of asthmatic guinea pigs was detected,and the bronchoalveolar lavage fluid(BALF) cells were counted.Lung tissue with HE and MCT immunohistochemical staining were used to observe the pathological changes in lung tissue and the distribution of MCT.Results:After injection of different concentration of acetylcholine chloride,the Re in group B and group C were increased significantly compared with group A(P<0.05):compared with group B.the Re in group C were decreased significantly(t=-5.385.-5.761.-6.184.-13.574.P<0.05):the total number of BALF cells and eosinophils were increased significantly in group B and C(t=19.618.9.598.10.854.5.388.P<0.05);compared with group B.the total number of BALF cells and eosinophils in group C was decreased significantly(t=-5.555.-5.392.P<0.05):the number of tryptase positive cells in group B was increased significantly than that in group A(t=21.312,P<0.05),and in addition to the alveolar septum and submucosa,the cells were also distributed around blood vessels and outside the cells:the number of tryptase positive cells in group C was decreased significantly compared with group B.and the difference was statistically significant(t=5.043.P<0.05).Conclusions:After the asthmatic guinea pigs arc treated with 1,25-dihydroxyvitamin D3,their BALF.Re.infiltration degree of inflammatory cells in the trachea and lung tissue and airway inflammatory reaction are reduced significantly.1,25-dihydroxyvitamin D3 has a certain inhibiting effect on the activation of mast cells and the release of MCT granules.

Possible biological and translational significance of mast cells density in colorectal cancer

Mast cells (MCs), located ubiquitously near blood vessels, are descended from CD34+ hematopoietic stem cells. Initially, although their role has been well defined in hypersensitivity reactions, the discovery of their sharing in both innate and adaptive immunity has allowed to redefine their crucial interplay on the regulatory function between inflammatory and tumor cells through the release of mediators granule-associated (mainly tryptase and vascular endothelial growth factor). In particular, in several animal and human malignancies it has been well demonstrated that activated c-Kit receptor (c-KitR) and tryptase (an agonist of the proteinase-activated receptor-2) take pivotal part in tumor angiogenesis after the MCs activation, contributing to tumor cells invasion and metastasis. In this review, we focused on crucial MCs density (MCD) role in colorectal cancer (CRC) development and progression angiogenesis-mediated; then, we will analyze the principal studies that have focused on MCD as possible prognostic factor. Finally, we will consider a possible role of MCD as novel therapeutic target mainly by c-KitR tyrosine kinase inhibitors (imatinib, masitinib) and tryptase inhibitors (gabexate and nafamostat mesylate) with the aim to prevent CRC progression.

Role of mast cell-mi R-490-5p in irritable bowel syndrome

AIM To determine the functional role of mi R-490-5p in mast cell proliferation and apoptosis,and in the mast cell tryptase/PAR-2 signal pathway.METHODS The 3rd generation of lentivirus vector systems containing enhanced green fluorescent protein(EGFP)(Ruisai Inc.,Shanghai,China),which acts as a reporter gene was used to construct the mmu-mi R-490-5p lentivirus expression vector p EGFP-antagomi R-490-5p,and the lentivirus vector p EGFP-negative was used as a negative control.The stably transfected mast cell line p815 was then constructed.GFP positive cells were successfully transfected cells.We determined the expression of mi R-490-5p in p815 mast cells before and after transfection using quantitative real-time PCR(q RT-PCR).In addition,after transduction with the lentivirus vectors,the role of mi R-490-5p in mast cell proliferation and apoptosis was investigated using the CCK-8 assay and flow cytometry,respectively.The m RNA levels of tryptase and PAR-2 were detected by q RT-PCR and the protein levels were detected by Western blot.RESULTS The inhibition of mi R-490-5p expression promoted apoptosis and inhibited proliferation of p815 mast cells.The m RNA levels of tryptase and PAR-2 were significantly increased after transfection comparedwith the control group,tryptase(P=0.721,normal vs null;P=0.001,si RNA vs normal;P=0.002,si RNA vs null)and PAR-2(P=0.027,si RNA vs null;P=0.353,normal vs null;P=0.105,si RNA vs normal).The protein levels of tryptase and PAR2 were slightly higher in the si RNA group than those in the control group,but the difference was not statistically significant(P>0.05).CONCLUSION mi R-490-5p plays a vital role in the pathogenesis of irritable bowel syndrome by affecting mast cell proliferation and apoptosis;with down-regulation of mi R-490-5p,the m RNA level of mast cell tryptase and PAR-2 increased,and the protein level increased,but the difference was not statistically significant.

A concise, practical guide to diagnostic assessment for mast cell activation disease

As recognition of mast cell(MC) involvement in a range of chronic inflammatory disorders has increased, diagnosticians' suspicions of MC activation disease(MCAD) in their chronically mysteriously inflamed patients have similarly increased. It is now understood that the various forms of systemic mastocytosis- diseases of inappropriate activation and proliferation of MCs seemingly driven by a small set of rare, usually constitutively activating mutations in assorted MC regulatory elements-comprise merely the tip of the MCAD iceberg, whereas the far larger and far more clinically heterogeneous(and thus more difficult to recognize) bulk of the iceberg consists of assorted forms of MC activation syndrome(MCAS) which manifest little to no abnormal MC proliferation and may originate from a far more heterogeneous set of MC mutations. It is reasonable to suspect MCAD when symptoms and signs of MC activation are present and no other diagnosis better accounting for the full range of findings is present. Initial laboratory assessment should include not only routine blood counts and serum chemistries but also a serum total tryptase level, which helps direct further evaluation for mastocytosis vs MCAS. Appropriate tissue examinations are needed to diagnose mastocytosis, while elevated levels of relatively specific mast cell mediators are sought to support diagnosis of MCAS. Whether assessing for mastocytosis or MCAS, testing is fraught with potential pitfalls which can easily yield false negatives leading to erroneous rejection of diagnostic consideration of MCAD in spite of a clinical history highly consistent with MCAD. Efforts at accurate diagnosis of MCAD are worthwhile, as many patients then respond well to appropriately directed therapeutic efforts.

Tryptase和TIM-1双阳性肥大细胞在不同程度人牙周炎组织中的量化研究

目的:T细胞免疫球蛋白黏蛋白1(T-cell immunoglobulin mucin 1,TIM-1)作为一种免疫调节分子,能影响肥大细胞的功能。TIM-1在牙周炎过程中是否在肥大细胞上表达未见报道。本研究旨在观察TIM-1在慢性牙周炎牙龈组织中肥大细胞上的表达,探讨类胰蛋白酶(tryptase)和TIM-1双阳性肥大细胞在牙周炎发病机制中的作用。方法:将92例接受研究的患者按慢性牙周炎的病变程度分成3组:(1)正常对照组27例;(2)轻度牙周炎组34例;(3)重度牙周炎组31例。牙龈组织标本经4%甲醛液固定48 h以上。牙龈标本经石蜡包埋、组织连续切片,HE染色,光学显微镜下观察牙龈的组织学改变;采用免疫荧光双染色,荧光显微镜下观察牙龈组织中tryptase和TIM-1双阳性肥大细胞的表达情况。结果:牙龈组织中的炎症反应程度与慢性牙周炎的病变程度趋势一致。与正常对照组相比较,轻度慢性牙周炎组(P<0.05)和重度慢性牙周炎组(P<0.01)牙龈组织中tryptase和TIM-1双阳性肥大细胞数量显著升高;重度牙周炎组牙龈组织中的tryptase和TIM-1双阳性肥大细胞数量高于轻度牙周炎组(P<0.05)。结论:慢性牙周炎牙龈组织中的tryptase和TIM-1双阳性肥大细胞数量与牙周炎程度的趋势相一致,提示tryptase和TIM-1双阳性肥大细胞可能参与对慢性牙周炎发病过程的免疫调节。

Tryptase阳性细胞在移植后不同时间段排斥肾中的分布

目的 了解Tryptase阳性肥大细胞在肾移植后不同时间段的排斥肾组织中的分布规律。方法 采用抗Tryptase单抗免疫组化染色法与甲苯胺蓝特殊染色方法 ,对移植后 8d～ 7a手术切除的排斥肾进行了染色。结果 在移植后的各个时间段的排斥肾中均见到较正常肾明显偏多的肥大细胞 ,且随着移植时间的延长呈增长的趋势。其中移植后 7a以上组的肥大细胞明显高于 1～ 2a组和 5～ 6a组 (P <0 .0 1)。甲苯胺蓝染色结果与Tryptase免疫组化结果相一致 ,但Tryptase免疫组化方法更为敏感。结论 肥大细胞可能参与了肾移植的排斥反应 。

肥大细胞类胰蛋白酶(MCT)在口腔鳞癌患者血清中表达水平的研究

目的:通过对比检测口腔鳞癌患者、良性颌面部肿瘤患者及健康对照者血清中的肥大细胞类胰蛋白酶(MCT)的表达水平,分析其在口腔鳞癌患者血清的表达水平有无异常,并分析其潜在的临床意义。方法:收集63例口腔鳞癌患者、47例颌面良性肿瘤患者及55例健康对照者的血清,用ELISA试剂盒检测血清中MCT的表达水平,后分析口腔鳞癌患者血清中MCT水平的临床意义。结果:口腔鳞癌患者血清中的MCT的表达水平高于其它两组,但不具有显著的统计学差异(P=0.09),而在口腔鳞癌组,MCT的浓度水平与肿瘤的大小呈正性相关,此外,在中分化鳞癌中MCT的表达水平相对较高。结论:MCT作为肥大细胞聚集、活化的标记物之一,在口腔鳞癌患者血清的表达并未显著升高,而血清MCT的表达水平与肿瘤大小有关。

通过MC-Tryptase-PAR-2-FLS途径探讨资木瓜总苷对K/B×N小鼠血清转移性关节炎的治疗作用

目的:研究资木瓜总苷(TSCS)通过MC-Tryptase-PAR-2-FLS途径对K/B×N小鼠血清转移性关节炎的治疗作用。方法:小鼠随机分为6组:C57BL/6小鼠分别注射C57BL/6小鼠血清、K/B×N小鼠血清作为正常对照组和模型组,KitW-4Bao小鼠注射K/B×N小鼠血清作为阴性对照组,移植肥大细胞(MC)的KitW-4Bao小鼠随机分为3组:阳性药物(色甘酸钠)组、TSCS低、高剂量组,均注射K/B×N小鼠血清。游标卡尺测量小鼠关节肿胀度,HE染色观察关节形态学变化,甲苯胺蓝染色观察MC脱颗粒变化,免疫组化和qRT-PCR检测关节组织中类胰蛋白酶(Tryptase)、蛋白酶激活受体2(PAR-2)表达情况,ELISA检测血清IgG及关节中炎症因子含量变化。结果:KitW-4Bao小鼠对血清转移性关节炎抵抗,移植MC后,对K/B×N小鼠血清恢复敏感,引发关节炎。与模型组相比,TSCS高剂量组小鼠关节肿胀度、关节炎评分、病理学评分、MC脱颗粒率、Tryptase和PAR-2表达、血清IgG和炎症因子水平显著降低。结论:MC在K/B×N小鼠血清转移性关节炎模型中发挥重要作用;TSCS通过抑制MC脱颗粒释放类胰蛋白酶,影响MC-Tryptase-PAR-2-FLS通路,治疗K/B×N小鼠血清转移性关节炎。

四逆散加减联合雷贝拉唑肠溶胶囊治疗胃食管返流性咳嗽患者的效果及对IL-8、SP、MCT水平的影响

目的探讨四逆散加减联合雷贝拉唑肠溶胶囊治疗胃食管返流性咳嗽的效果及对患者白细胞介素-8（interleukin 8,IL-8）、P物质（substance P,SP）和肥大细胞类胰蛋白酶（mast cell tryptase,MCT）水平的影响。方法 92例胃食管返流性咳嗽患者按照抽签法分为对照组与实验组,每组各46例。对照组采用雷贝拉唑肠溶胶囊治疗,实验组在对照组基础上加用四逆散加减治疗,比较2组疗效,咳嗽症状积分,IL-8、SP、MCT,胃动素（gastric dynamic element,MOT）及胃泌素（gastrin-releasing,GAS）,肺功能和安全性。结果实验组总有效率（93.47%）高于对照组（76.08%）,比较差异有统计学意义（P〈0.05）。治疗后,实验组咳嗽症状积分（1.53±0.19）分高于对照组（1.96±0.24）分,比较差异有统计学意义（P〈0.05）。实验组IL-8、SP、MCT水平均明显低于对照组（P〈0.05）。实验组MOT、GAS、肺功能较对照组改善更明显（P〈0.05）。2组安全性比较差异无统计学意义。结论四逆散加减联合雷贝拉唑肠溶胶囊治疗胃食管返流咳嗽的效果确切,可降低患者IL-8、SP、MCT水平。

Close relationship between mediators of inflammation and pancreatic cancer:Our experience

In this editorial,we focus specifically on the mechanisms by which pancreatic inflammation affects pancreatic cancer.Cancer of the pancreas remains one of the deadliest cancer types.The highest incidence and mortality rates of pancreatic cancer are found in developed countries.Trends of pancreatic cancer incidence and mortality vary considerably worldwide.A better understanding of the etiology and identification of the risk factors is essential for the primary prevention of this disease.Pancreatic tumors are characterized by a complex microenvironment that orchestrates metabolic alterations and supports a milieu of interactions among various cell types within this niche.In this editorial,we highlight the foundational studies that have driven our understanding of these processes.In our experimental center,we have carefully studied the mechanisms of that link pancreatic inflammation and pancreatic cancer.We focused on the role of mast cells(MCs).MCs contain pro-angiogenic factors,including tryptase,that are associated with increased angiogenesis in various tumors.In this editorial,we address the role of MCs in angiogenesis in both pancreatic ductal adenocarcinoma tissue and adjacent normal tissue.The assessment includes the density of c-Kit receptor-positive MCs,the density of tryptase-positive MCs,the area of tryptasepositive MCs,and angiogenesis in terms of microvascularization density.

肾癌组织中肥大细胞脱颗粒特征及其与血管形成的关系

目的探讨肾癌组织中肥大细胞脱颗粒特征及其与肿瘤血管形成的关系。方法通过类胰蛋白酶(Tryptase)免疫组织化学染色标记人肾癌组织及癌旁肾组织中的肥大细胞并统计其浸润密度,根据Tryptase在细胞内外的分布判断肥大细胞脱颗粒状态;通过CD31免疫组织化学染色标记血管内皮细胞并计数肿瘤微血管密度。比较肾癌组织与癌旁肾组织中肥大细胞数量、脱颗粒水平差异,分析肾癌组织中肥大细胞脱颗粒水平与其浸润密度及肿瘤微血管密度的关系。结果共纳入125例肾癌组织标本和52例癌旁肾组织标本。Tryptase染色分析发现肾癌组织和癌旁肾组织中肥大细胞密度分别为(2.67±0.22)、(0.63±0.14)个/高倍视野,肥大细胞脱颗粒阳性率分别为70.3%、34.3%,差异有统计学意义(P<0.01)。肥大细胞密度低、中、高三组的肥大细胞脱颗粒阳性率分别为42.1%、64.1%和83.3%。肥大细胞脱颗粒阳性组肿瘤微血管密度明显高于脱颗粒阴性组[(24.18±1.64)个/高倍视野vs.(10.18±1.97)个/高倍视野,P<0.01]。结论肥大细胞在肾癌组织中的脱颗粒水平明显高于癌旁肾组织,且其脱颗粒水平与肥大细胞浸润数量及肿瘤血管形成密切相关。

Early development and functional properties of tryptase/chymase double-positive mast cells from human pluripotent stem cells

Mast cells (MCs) play a pivotal role in the hypersensitivity reaction by regulating the innate and adaptive immune responses. Humans have two types of MCs. The first type, termed MCTC, is found in the skin and other connective tissues and expresses both tryptase and chymase, while the second, termed MCT, which only expresses tryptase, is found primarily in the mucosa. MCs induced from human adult-type CD34+ cells are reported to be of the MCT type, but the development of MCs during embryonic/fetal stages is largely unknown. Using an efficient coculture system, we identified that a CD34+c-kit+ cell population, which appeared prior to the emergence of CD34+CD45+ hematopoietic stem and progenitor cells (HSPCs), stimulated robust production of pure Tryptase+Chymase+ MCs (MCTCs). Single-cell analysis revealed dual development directions of CD34+c-kit+ progenitors, with one lineage developing into erythro-myeloid progenitors (EMP) and the other lineage developing into HSPC. Interestingly, MCTCs derived from early CD34+c-kit+ cells exhibited strong histamine release and immune response functions. Particularly, robust release of IL-17 suggested that these early developing tissue-type MCTCs could play a central role in tumor immunity. These findings could help elucidate the mechanisms controlling early development of MCTCs and have significant therapeutic implications.

肥大细胞蛋白酶在过敏性哮喘中的作用

肥大细胞(Mast cells,MC)在过敏性哮喘中起着重要作用,在过敏原的刺激下,MC通过Fc?RI结合的IgE抗体识别过敏原而被激活后,释放大量介质,其中释放的大量蛋白酶,包括类胰蛋白酶、类糜蛋白酶,对哮喘的进展起重要作用,既可保护气道、防止气道重塑,又可促进炎症发生、聚集炎症细胞、破坏气道等。本文综述了MC蛋白酶在过敏性哮喘发生发展中的最新研究进展,围绕MC蛋白酶在气道上皮细胞、平滑肌细胞、成纤维细胞中的分子机制进行了总结和讨论,以期为哮喘的诊断和治疗提供一定的参考。

针刺对佐剂性关节炎大鼠滑膜肥大细胞脱颗粒和类胰蛋白酶表达的影响

目的:观察针刺对佐剂性关节炎大鼠滑膜组织病理改变、滑膜肥大细胞脱颗粒及类胰蛋白酶表达的影响,探讨肥大细胞功能与针刺治疗大鼠佐剂性关节炎的关系。方法:46只雄性Wistar大鼠随机分为正常对照组(n=16)、模型组(n=15)和针刺组(n=15)。除正常对照组外,其余大鼠通过足垫部注射0.1mL弗氏完全佐剂建立佐剂性关节炎模型。针刺组隔天针刺双侧足三里、悬钟、肾俞穴,每次15min,共8次。治疗期间,正常对照组和模型组除不予针刺外,其余处理均同针刺组。设造模当天为实验第0天,之后每3天对大鼠的体质量和足跖体积进行测量并记录。治疗结束后,取所有大鼠右后踝关节滑膜组织,HE染色后根据滑膜病理5级评分法评价滑膜组织病理改变,甲苯胺蓝染色法观察滑膜肥大细胞数量及其脱颗粒情况,免疫组织化学法检测类胰蛋白酶在滑膜组织中的表达情况。结果:与模型组比较,针刺组大鼠治疗后体质量明显增加(P<0.05),足跖体积明显减小(P<0.01)。HE染色显示,针刺可显著减轻佐剂性关节炎大鼠滑膜组织中的炎细胞浸润、滑膜细胞增生和滑膜纤维组织增生等病理表现(P<0.05);甲苯胺蓝染色显示,与正常对照组比较,模型组大鼠滑膜肥大细胞数量和脱颗粒率均明显增加(P<0.01),而针刺组大鼠滑膜肥大细胞数量和脱颗粒率的上升得到有效抑制,与模型组比较差异有统计学意义(P<0.01);免疫组织化学检测显示,针刺可显著抑制佐剂性关节炎大鼠滑膜组织中上调的类胰蛋白酶表达(P<0.01)。相关分析显示,滑膜肥大细胞数量及脱颗粒率均与滑膜组织病理改变总评分呈明显的正相关(P<0.01)。结论:针刺具有治疗大鼠早期佐剂性关节炎和调节滑膜肥大细胞功能的作用,由于两者间存在明显的正相关性,因此针刺可能通过抑制滑膜肥大细胞功能,对佐剂性关节炎大鼠起治疗作用。

肥大细胞在湿热型溃疡性结肠炎大鼠的表达与黄芩汤的调节

目的探讨肥大细胞(MC)在湿热型溃疡性结肠炎(UC)发病机制中的作用以及黄芩汤对湿热型UC模型中MC的调节。方法采用高脂高糖饲料联合高温高湿人工气候模拟法造出湿热型体质,结合三硝基苯磺酸灌肠法诱发UC最终建立湿热型UC大鼠模型;造模成功后将大鼠分为模型组(n=15)、黄芩汤组(n=13)及美沙拉嗪组(n=12),灌胃治疗1周后,采用HE染色法观察病变部位炎症细胞浸润程度,甲苯胺蓝改良染色法检测MC形态、数目及脱颗粒率,免疫组化SP法检测类胰蛋白酶(TA),ELISA法检测血清IL-6。结果与空白对照组(n=15)相比,模型大鼠结肠病理切片炎症浸润程度、MC数目、TA、IL-6明显升高(P<0.05);治疗后黄芩汤及美沙拉嗪组以上指标均较模型对照组降低(P<0.05),且两种药物的效应无明显组间差异(P>0.05)。结论 MC等肠粘膜局部免疫细胞在湿热型UC发病机制中发挥着重要作用;黄芩汤可降低炎症浸润程度,减少MC数目,降低TA的释放,抑制IL-6的分泌,此可能为黄芩汤治疗湿热型UC的机制。

豚鼠过敏性休克类胰蛋白酶活力测定

目的探索过敏性休克法医学客观诊断标准。方法建立豚鼠异种血清过敏性休克模型,采用专性底物对模型鼠的血清,肺,气管类胰蛋白酶活力进行测定。结果过敏性休克豚鼠血清,肺,气管类胰蛋白酶活力均增加,且三者增加的程度是平行的。结论过敏性休克时,血清,肺,气管类胰蛋白酶活力增加可作为法医检案客观诊断标准。

蛋白酶抑制剂对肥大细胞类胰蛋白酶分泌的影响

目的 探讨蛋白酶抑制剂和组胺对肥大细胞类胰蛋白酶分泌的影响。方法 扁桃体组织经酶消化后 ,细胞成份用全HBSS重新悬浮。肥大细胞激发和抑制剂作用的试验在 37℃条件下完成。类胰蛋白酶水平用酶联免疫吸附试验(ELISA)方法测定。结果 鱼精蛋白具有刺激人类扁桃体肥大细胞释放类胰蛋白酶的作用 ,其分泌量可高达基础分泌量的5倍。高浓度TLCK和TPCK可抑制抗 IgE诱导的类胰蛋白酶释放。TLCK在有否预培养的情况下均能抑制钙离子导入剂 (cal ciumionophore ,CI)诱导的类胰蛋白酶释放 ,而TPCK则只有在 2 0min预培养后才能显示此作用。结论 TLCK和TPCK抑制IgE依赖性和非依赖性类胰蛋白酶释放提示具有胰蛋白酶和糜蛋白酶活性的酶参与肥大细胞的激活 分泌偶联过程。