肥大细胞在动脉粥样硬化形成中的作用

动脉粥样硬化,是冠心病的病理基础,被认为是一种慢性炎症性疾病,涉及如巨噬细胞和T淋巴细胞等许多炎性细胞。肥大细胞是一种重要的免疫细胞,其功能主要是在超敏反应方面的作用。有病理学研究表明：肥大细胞在动脉粥样硬化斑块周围表达增加,这表明肥大细胞可能与疾病的进展有关。最近的研究表明,肥大细胞在动脉粥样硬化中确实起着重要的作用。本文通过总结肥大细胞在动脉粥样硬化形成中的作用,为在疾病进程中,通过调节肥大细胞功能来改善动脉粥样硬化的这种治疗方式的可能性提供依据。     

肥大细胞在动脉粥样硬化形成中的作用

动脉粥样硬化,是冠心病的病理基础,被认为是一种慢性炎症性疾病,涉及如巨噬细胞和T淋巴细胞等许多炎性细胞。肥大细胞是一种重要的免疫细胞,其功能主要是在超敏反应方面的作用。有病理学研究表明:肥大细胞在动脉粥样硬化斑块周围表达增加,这表明肥大细胞可能与疾病的进展有关。最近的研究表明,肥大细胞在动脉粥样硬化中确实起着重要的作用。本文通过总结肥大细胞在动脉粥样硬化形成中的作用,为在疾病进程中,通过调节肥大细胞功能来改善动脉粥样硬化的这种治疗方式的可能性提供依据。     

Impacts of peripheral obestatin on colonic motility and secretion in conscious fed rats

Obestatin, a novel putative 23-amino acid peptide, was found to be derived from a mammalian preproghrelin gene by using a bioinformatics approach. Although the effects of obestatin on food intake and upper gut motility remain controversial, no studies have been carried out to explore its influence on lower gut motility and secretion. We investigated the impacts of intravenous (IV) injection of obestatin on rat colonic motor and secretory functions. Colonic transit time, fecal pellet output, and fecal content were measured in freely fed, conscious rats, which were chronically implanted with IV and colonic catheters. To test the validity of this animal model, human/rat corticotropin-releasing factor (h/rCRF) served as a stimulatory inducer of colonic motility and secretion. IV injection of obestatin (45, 100, and 300nmol/kg) did not affect the colonic transit time, whereas IV injection of h/rCRF (30nmol/kg) effectively accelerated colonic transit time. IV obestatin, in every dose we tested, also did not modify fecal pellet output, frequency of watery diarrhea, total fecal weight, fecal dried solid weight, or fecal fluid weight in the first hour after injection. On the other hand, IV injection of h/rCRF significantly enhanced fecal pellet output, as well as increased the frequency of watery diarrhea, total fecal weight, fecal dried solid weight, and fecal fluid weight during the first hour after injection compared with IV saline controls. In conclusion, peripheral obestatin administration has no impact on colonic motility and secretion in conscious fed rats.     

Protein kinase C alpha, betaI, and betaII isozymes regulate cytokine production in mast cells through MEKK2/ERK5-dependent and -independent pathways

Regulation of MAPK pathways by PKC isoforms was examined in murine bone marrow-derived mast cells (BMMCs). The PKCalpha, betaI, and betaII isoforms showed the most robust activation after FcepsilonR1-mediated stimulation by anti-ovalbumin specific IgE and ovalbumin (IgE-ova). PKCalpha, betaI, and betaII were all involved in activation of JNK, MEKK2, and ERK5, with differential relative contributions of each isoform to specific MAPK pathway components. BMMCs from mice lacking MEKK2 showed reduced production (50-60%) of IL-6, IL-13, and TNF-alpha after stimulation, demonstrating MEKK2-dependent and -independent pathways for cytokine production. Cytokine production was stimulated by over-expression of PKC in cells from MEKK2-deficient and wild-type mice. Activation of ERK5 did not occur in BMMCs lacking MEKK2, indicating that MEKK2-independent cytokine production was also ERK5-independent. Since MAPK modules differentially regulate mast cell functions, including degranulation and cytokine production, it is suggested that specific functions could be targeted by inhibiting specific PKC isoforms.     

Synthesis and pharmacological evaluation of the individual stereoisomers of 3-[methyl(1,2,3,4-tetrahydro-2-naphthalenyl)amino]-1-indanone, a potent mast cell stabilising agent

Each stereoisomer of 3-[methyl(1,2,3,4-tetrahydro-2-naphthalenyl)amino]-1-indanone, 1a-d, was prepared and evaluated in vitro for its ability to prevent mediator release induced by different degranulating agents from rodent mast cells and also in vivo against passive cutaneous anaphylaxis. The manner in which the stereoisomers prevented direct membrane activation was found to be highly dependent on the stereochemistry of the individual isomers. Stereoisomer 1b was the most active isomer in vivo, exhibiting superior activity to disodium cromoglycate.     

儿童功能性便秘肠道传输功能与肠道微生态的关系

目的 采用不透光X线标志物测定结肠传输指数(TI)对功能性排便障碍进行分型,探讨各型功能性便秘患儿之间及其与健康儿童之间肠道菌群差异。 方法 选取功能性便秘儿童40例,进行结肠传输试验并进行分型,其中33例出口梗阻型(出口梗阻组),7例慢传输型(慢传输组)。留取便秘儿童新鲜粪便样本,同时收集20例正常儿童(对照组)粪便标本,存放于-80 ℃冰箱。采用16S rRNA测序技术鉴定菌群。 结果 菌群多样性分析:出口梗阻组、慢传输组与对照组粪便标本在丰富度上差异存在统计学意义(P结论 出口梗阻型与慢传输型相比,有部分菌种在慢传输型便秘儿童中明显减少,这需要进一步研究论证;便秘儿童肠道菌群与健康人群存在较大差异,改变肠道菌群可能成为治疗慢性功能性便秘的新策略。慢传输型便秘病例数较少,需增加样本量后进一步分析。     

肥大细胞在大鼠肝癌发生过程中的作用研究

应用组织化学、免疫组织化学、电子显微镜及形态测量等方法对二乙基亚硝胺（ＤＥＮ）诱发大鼠肝肿瘤过程中肥大细胞（ＭＣ）对肝细胞生化代谢、细胞核及核仁组成区（ＮＯＲ）的影响等进行了研究。结果显示肝细胞葡萄糖６磷酸酶（Ｇ－６－Ｐａｓｅ）和α－抗胰蛋白酶（α－ＡＴ）在ＭＣ最多与最少组之间的分布无明显差异,而嗜银蛋白颗粒（ＡｇＮＯＲ）计数和纤维中心数密度及核仁面积之比则自第十四周起显示出分布的个体差异,它们在ＭＣ最多组明显低于ＭＣ最少组、本文并对该种差异的意义进行了简要讨论。     

尼罗罗非鱼消化道肥大细胞的组化性质

实验采用改良甲苯胺蓝(MTB)、阿利新蓝-沙黄(AB/SO)、甲基绿-派洛宁(MG-P)、天青Ⅱ-伊红-瑞氏混合液和硫堇5种组化染色法,对尼罗罗非鱼(Nile tilapia)消化道组织中的肥大细胞(Mast cell,MC)组化性质进行研究。尼罗罗非鱼的食管、胃及小肠壁内均显示有肥大细胞,在食管和胃的切片标本上肥大细胞主要分布在黏膜固有层和胃腺体之间。在肠道中的肥大细胞主要分布在黏膜固有层和肠上皮下方,少量肥大细胞存在于黏膜下层结缔组织中。细胞呈圆形、椭圆形,也有长梭形的。而且肥大细胞有沿血管分布的特点。5种组化染色结果表明:AB/SO、MTB和MG-P显示的MC效果较好,尤其AB/SO染色效果最好,肥大细胞轮廓清楚,胞质颗粒较清晰;尼罗罗非鱼肥大细胞胞浆颗粒都呈红色,即肥大细胞胞浆主要含肝素,不含组胺。天青Ⅱ-伊红-瑞氏混合液染色效果也很好,但被染的肥大细胞较少;80%乙醇硫堇染色,在尼罗罗非鱼消化道各段组织中均未能鉴定出肥大细胞。尼罗罗非鱼消化道肥大细胞大多分布于浅层的黏膜或血管、腺体周围的结缔组织等易表露于环境抗原的位点。罗非鱼消化道黏膜层结缔组织中的肥大细胞与大多数脊椎动物的肥大细胞一样,具有沿血管分布的特性,说明硬骨鱼的肥大细胞如哺乳动物肥大细胞一样与血管有着密切的关系。     

Mouse mast cell protease-1 cleaves angiotensin I to form angiotensin II

The ability to convert angiotensin (Ang) I to Ang II was compared between human alpha-chymase and two mouse beta-chymases, mouse mast cell protease (mMCP)-1 and mMCP-4. Human chymase hydrolyzed Ang I to produce Ang II without further degradation. mMCP-1 similarly generated Ang II from Ang I in a time-dependent manner and the formation of the fragment other than Ang II was marginal. In contrast, mMCP-4 hydrolyzed Ang I at two sites, Tyr(4)-Ile(5) and Phe(8)-His(9), with Ang II formation being tentative. Consistently, mMCP-4 but not human chymase hydrolyzed Ang II and mMCP-1 showed little hydrolytic activity against Ang II. These data suggest that not only human chymase but also mMCP-1 might possess a physiological role in Ang II formation. Our findings also imply that the Ang-converting activity of chymase may not be related to the categorization of chymase into alpha- or beta-type based on their primary structure.     

Cellular and extracellular changes following mast-cell secretion in avascular rat mesentery

Summary Mast cell (MC) secretion induces local cell proliferation lasting 48–72 h in fibroblasts and mesothelial cells in the almost avascular true mesentery of the rat. We studied this membranous tissue by transmission electron microscopy with regard to cellular and extracellular features occurring during the first 72 h following MC secretion.After MC secretion elicited by compound 48/80, apparently all individual tissuebound cells (i.e. fibroblasts, mesothelial cells, and macrophages) show signs of accelerated metabolic activity. In fibroblasts, conspicuous increases in the volume of Golgi apparatus and rough endoplasmic reticulum and in the amount of plasmalemmal indentations suggest an increased production and secretion of the extracellular matrix. Released MC granules lying close to projections of nearby phagocytosing cells cause areas free from electrondense material in the extracellular matrix. MC secretion therefore appears to produce a remodelling of extracellular matrix. Most of the activities initiated by MC secretion start to subside within (48-)72 h.The findings indicate a close functional relationship between the tissue MC and all its neighbouring cells and the surrounding extracellular matrix. The striking chain of events that it induces emphasizes strongly that the secreting MC plays a prominent although as yet in many respects enigmatic role in normal tissue.Key to Abbreviations CM cytoplasmic matrix - GA Golgi apparatus - MC mast cell - PL primary lysosome - RER rough endoplasmic reticulum - TEM transmission electron microscopySupported by grants from the Swedish Medical Research Council, Project 5942     

Über die Entstehung von Mastzellgranula

Zusammenfassung In den Mastzellen der Gallenblase fetaler und neugeborener Meerschweinchen kommen neben wenigen homogenen Körnchen reichlich grobkörnige, reticuläre und lamelläre Granula vor. Da der Golgi-Apparat nur schwach entwickelt ist und nicht alle Granula von Membranen umgeben sind, wird geschlossen, daß die spezifischen Granula nicht in dieser Zellorganelle entstehen. Es gibt Anzeichen dafür, daß sie sich in situ im Zytoplasma entwickeln.

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The genesis of mast cell granules

Summary The mast cells in the gall bladder of fetal and newborn guinea-pigs contain few homogeneous granules and many coarse-granular, reticular and lamellate ones. Since the Golgi apparatus is poorly developed and since not all the granules are membrane bound, it is concluded that the specific granules do not originate in this organelle. There are indications that the granules develop in situ in the cytoplasm.

     

Rapid degradation of neurotensin by stimulated rat mast cells

A RIA towards neurotensin (NT) using C-terminal- and N-terminal-specific antisera was used to study degradation of this tridecapeptide by isolated rat mast cells. Incubation of NT (10 μM) with peritoneal or pleural mast cells resulted in a rapid loss of NT immunoreactivity (iNT), as measured by C-terminal-directed antiserum, with little effect on N-terminal iNT. The rate of the reaction was faster with pleural cells (T_1/2, 30 s) than with peritoneal cells (T_1/2, 180 s) and was > 10-fold slower in the presence of metabolic poisons. The enzyme(s) involved is most likely released from the cells during secretion, as NT was degraded by media conditioned by compound 48/80-stimulated mast cells 40–60 times faster than by media from unstimulated cells. This degradation by conditioned media was concentration dependent, pH dependent, and temperature sensitive. HPLC analyses indicated a near stoichiometric conversion of NT to NT(1–12) (66%) and NT(1–11) (34%) after incubation for 10–30 s with conditioned media. By 30 min only NT(1–11) and NT(1–10) were present. Phenanthroline (1 mM), an inhibitor of carboxypeptidase, prevented the loss of C-terminal iNT and the generation of NT(1–12) and NT(1–11). While NT(1–12) was effective in releasing histamine from mast cells in vitro and increasing vascular permeability in vivo, NT(1–11) was not. These results suggest that carboxypeptidase-like enzyme(s) could modulate the level and form of NT-related peptides in various states involving activation of mast cells.     

新生大鼠雌激素注射后睾丸肥大细胞的变化

新生大鼠注射雌二醇后,睾丸肥大细胞于第30天可见到,细胞数量随年龄增长而增多,生后4－6个月,睾丸网附近仍可见大量肥大细胞。睾丸内的肥大细胞比皮肤内的结缔组织肥大细胞（CTMC）小而与小肠粘膜的粘膜肥大细胞（MMC）相近,AB－S染色后基本着蓝色,硫酸小檗碱荧光染色后呈现中等强度黄色荧光,结果提示,新生大鼠雌激素注射后睾丸内肥大细胞的增多可能与免疫过程有关,睾丸内肥大细胞与CTMC和MMC皆有所不同。     

Induction of histamine release from rat mast cells by bradykinin analogues

Independently of their agonistic or antagonistic activity on different isolated tissue preparations, the kinin analogues investigated induce histamine release on rat peritoneal mast cells. The effectivity of most compounds is 10 to 100 times higher than that of bradykinin. Beside the positively charged amino acids, the elongation at the N-terminus with hydrophobic amino acids and the replacement of amino acids in the bradykinin sequence (especially at position 7) with aromatic residues is important for a high histamine-releasing activity.     

卷曲乳杆菌对三种结肠上皮细胞的粘附性

目的 研究卷曲乳杆菌对3种结肠上皮细胞（SW480细胞、SW620细胞、LOVO细胞）的粘附性。方法 将处于对数生长期的卷曲乳杆菌A7分别与SW480细胞、SW620细胞、LOVO细胞进行体外粘附试验,革兰染色后显微镜观察卷曲乳杆菌A7对3种结肠上皮细胞的粘附结果并计数。结果 卷曲乳杆菌A7对3种结肠上皮细胞的粘附均具有显著性,其中对于SW480细胞和LOVO细胞的粘附性明显高于SW620细胞。结论 卷曲乳杆菌A7对SW480细胞、SW620细胞、LOVO细胞均具有较强的粘附性,提示该菌株有望成为肠道益生菌的新成员。     

甘氨酸偶联皮质醇抗Compound 48/80刺激大鼠急性过敏反应的作用研究

目的:观察甘氨酸偶联皮质醇(HG)对Compound 48/80(C48/80)刺激大鼠急性过敏反应的影响,以期为后期深入探讨糖皮质激素非基因组机制建立有效研究工具。方法:将60只雄性SD大鼠随机分为4组:急性过敏模型组(腹腔注射C48/80+对照溶剂,n=15)、皮质醇(GC)干预组(腹腔注射C48/80+GC,n=15)、HG干预组(腹腔注射C48/80+HG,n=15)、空白对照组(腹腔注射等量生理盐水,n=15)。造模组及各干预组分别在腹腔注射C48/80 5 min后分别给予腹腔注射对照溶剂(等量无水乙醇,为GC及HG的稀释溶剂)、GC或HG,之后每5 min监测大鼠肛温及平均动脉压(MAP),腹腔注射C48/80后30 min时处死大鼠,取大鼠咽喉组织及肺组织,多聚甲醛固定切片后观察组织形态;同时摘眼球取血,分离血清,采用酶联免疫吸附实验(ELISA)测定血清中组胺含量。结果:与空白对照组相比,造模组大鼠肛温及平均动脉压呈持续降低趋势,C48/80刺激可诱发大鼠咽喉及肺组织水肿、肺泡损坏及炎性细胞浸润,大鼠血清中组胺水平较对照组显著升高(P0.05);较造模组各干预组均可减轻造模诱发的上述过敏反应(P0.05),但HG干预组较GC干预组促进大鼠肛温降低恢复起效时间快、复温效率高(P0.05),HG干预组减弱大鼠平均动脉压持续性降低趋势较GC干预组效果明显(P0.05);HG干预组显著减轻C48/80刺激导致的咽喉及肺组织水肿、肺泡损坏及炎性细胞浸润;HG干预组能显著抑制C48/80刺激导致的血清组胺升高水平(P0.05),但HG干预组与GC干预组抑制效果无明显差异(P0.05)。结论:HG能够快速抑制C48/80刺激诱导的大鼠急性过敏反应;能够显著抑制肥大细胞组胺的释放;HG可能通过非基因组作用快速抑制肥大细胞的脱颗粒,抑制炎性介质组胺的释放,从而抑制免疫炎症反应,发挥抗过敏及抗炎症效应,从而为研究糖皮质激素非基因组机制提供了有效的研究工具。     

Human mast cells express intracellular TRAIL

Recently we demonstrated that human mast cells (MC) express functional TRAIL death receptors. Here we assessed the expression of TRAIL on both mRNA and protein level in cord blood derived MC (CBMC) and HMC-1. The TRAIL release either spontaneous or induced by LPS, IFN-γ and IgE-dependent activation, was evaluated as well. The protein location was restricted to the intracellular compartment in CBMC, but not in HMC-1. The intracellular TRAIL was not localized inside the granules. The treatment with IFN-γ and LPS up-regulated intracellular TRAIL expression in CBMC, but did not induce its release. These in vitro data show that human MC can produce and express intracellular TRAIL whose location could not be altered by different stimuli.