天花粉对小鼠子宫肥大细胞超微结构的影响

本研究用雌性中国１号小鼠１２只,分实验组和对照组,实验组分别腹腔注射天花粉后处死,取子宫组织作超薄切片,电镜观察。在天花粉作用下,常见子宫肥大细胞脱颗粒,呈功能活化状态,并与子宫平滑肌细胞紧密相邻。提示肥大细胞可能通过脱颗粒释放组胺促进子宫平滑肌的收缩。同时,也见肥大细胞与子宫结缔组织中的成纤维细胞,淋巴细胞等密切接触,提示成纤维细胞和淋巴细胞与天花粉作用下子宫肥大细胞的数量增多有关。     

The role of mast cells in thioglycollate-induced inflammation

The possible role of mast cells in the initiation of inflammation was studied in genetically mast cell-deficient mice, WBB6F1-W/Wv. Inflammation was induced by i.p. injection of thioglycollate. The influx of neutrophils was markedly delayed in WBB6F1-W/Wv mice as compared to the WBB6F1-+/+, mice (congeneic controls). At the time (14 h) of maximum influx of neutrophils in WBB6F1-+/+ mice, thioglycollate caused a 3-fold increase in the total cell number in the peritoneal lavage fluid, and the neutrophil count was elevated 14-fold. At the same time point in W/Wv mice, the total cell number in the peritoneal lavage fluid was not increased significantly and the neutrophils were increased only three- to four-fold. Not only was the neutrophil influx in WBB6F1-W/Wv mice delayed, but the length of time during which the neutrophil count was elevated in the peritoneal fluid was significantly shortened. Transfer (i.p.) of mast cells cultured from the bone marrow of congeneic controls corrected the delay in the neutrophil influx. The magnitude of the neutrophil influx in WBB6F1-W/Wv mice was equivalent to that of congeneic controls 9 days after mast cell repletion. Histologic studies were performed to follow the migration and differentiation of mast cells after adoptive transfer into WBB6F1-W/Wv mice. No connective tissue mast cells could be identified on day 9 when the inflammatory reaction was restored. Migration of mast cells into the tissue, as studied in the cecum, progressed steadily. On day 9 after adoptive transfer, the mast cell number was 38% of congeneic controls. Therefore, the increase in thioglycollate-induced neutrophil influx in WBB6F1W/Wv mice after mast cell repletion seemed to be correlated, at least to some extent, with the migration of mast cells into tissues and not with differentiation into connective tissue mast cells. However, a certain maturation and differentiation may have occurred. These results suggest that mast cells play an important role, although they do not seem to be the only cell type responsible for the initiation of inflammation.     

Human uterine mast cells. Isolation, purification, characterization, ultrastructure, and pharmacology.

As part of an ongoing investigation of human mast cell heterogeneity, we have isolated, partially purified, and characterized the uterine mast cell and compared it with mast cells isolated from other organs. The average histamine content of myometrium and leiomyofibroma obtained from hysterectomies was 2.1 +/- 0.3 (mean +/- SEM) microgram/g of tissue (n = 10), and the histamine content of the two tissues did not differ significantly. A mild collagenase, hyaluronidase, and DNase digestion was used to disperse the uterine mast cells, with an average yield of 9.5% (range, 0 to 21%). The average histamine/uterine mast cell was 2.1 +/- 0.2 pg (n = 3), and 61 +/- 7% (n= 3) of the uterine mast cells survived overnight culture. Early purification efforts with Percoll gradients have yielded up to 80% pure uterine mast cells, with an average of 27 +/- 10% (n = 5). Uterine mast cells released histamine in response to the secretogogues anti-IgE and A23187 but did not respond to substance P or to the basophil secretogogues FMLP, C5a, and 12-O-tetradecanoylphorbol-13-acetate. After 1 microgram/ml anti-IgE stimulation, the uterine mast cell appeared to make significant quantities of PGD2 (89 +/- 26 ng/10(6) cells, n = 6) (p less than 0.05), as assayed by RIA. Simultaneously, leukotriene C4 release was 45 +/- 15 ng/10(6) cells, (n = 6) (p less than 0.05), as assayed by RIA. Combined gas-chromatography mass spectroscopy analysis of anti-IgE-stimulated cell supernatants confirmed the production of PGD2. In pharmacologic studies, isobutyl-methylxanthine and isoproterenol blocked anti-IgE-induced histamine release. The uterine mast cell is similar to the lung mast cell in terms of response to secretogogues and release of arachidonic acid metabolites. Ultrastructurally, the uterine mast cell contains scroll granules, crystal granules, combined granules, homogeneously dense granules, and large lipid bodies, many with focal lucencies within them. Particle granules, most frequently present in gut mast cells of mucosal origin, were absent from uterine mast cells. Although certain features are analogous to the ultrastructure of skin or lung mast cells, the combination of structures is distinctive for uterine mast cells.     

天花粉注射后不同时间小鼠子宫肥大细胞的变化

雌性中国１号小鼠３６只,分为实验组和对照组。实验组分别腹腔注射天花粉后不同时间处死,取子宫组织,作甲苯胺蓝及Ａｌｃｉａｎ蓝,藏红染色,对肥大细胞进行计数后结果表明：给小鼠天花粉,可引起其子宫肥大细胞数量明显增多,而且随天花粉作用时间延长,肥大细胞数量逐渐增多。本文对肥大细胞的这一数量变化与子宫功能的关系也作了进一步探讨。     

输卵管妊娠时输卵管壁肥大细胞的研究

目的　探讨肥大细胞在输卵管妊娠中的数量变化及其与血清性激素的关系。方法 :取输卵管妊娠时的输卵管及月经周期的增生期、分泌期和正常宫内早孕时的输卵管 ,常规石蜡切片 ,用甲苯胺蓝染色法显示肥大细胞 ;用酶免疫分析法检测输卵管妊娠患者、正常育龄未孕妇女 (增生期和分泌期 )及正常宫内早孕妇女血清雌二醇和孕酮水平。结果 :输卵管妊娠患者血清雌二醇和孕酮水平均高于正常育龄未孕妇女 (增生期和分泌期 ) ,低于正常宫内早孕妇女 ,四组间两两比较差异均有显著性 (P <0 0 5 ) ;肥大细胞主要分布于输卵管肌层 ,其数量变化为 :输卵管妊娠组较增生期和分泌期这两组均少 ,差异均有显著性 (P <0 0 5 ) ,而增生期和分泌期这两组肥大细胞数量变化不明显 ,差异无显著性 (P >0 0 5 ) ;两例正常宫内早孕时的输卵管壁肥大细胞数量明显比输卵管妊娠组多 ,与增生期和分泌期这两组比较 ,肥大细胞数量变化不明显。结论 :1 人输卵管壁内肥大细胞的数量不受血清性激素水平的影响。 2 输卵管妊娠时肥大细胞数量减少     

Relationship between Numerous Mast Cells and Early Follicular Development in Neonatal MRL/MpJ Mouse Ovaries

In the neonatal mouse ovary, clusters of oocytes called nests break into smaller cysts and subsequently form individual follicles. During this period, we found numerous mast cells in the ovary of MRL/MpJ mice and investigated their appearance and morphology with follicular development. The ovarian mast cells, which were already present at postnatal day 0, tended to localize adjacent to the surface epithelium. Among 11 different mouse strains, MRL/MpJ mice possessed the greatest number of ovarian mast cells. Ovarian mast cells were also found in DBA/1, BALB/c, NZW, and DBA/2 mice but rarely in C57BL/6, NZB, AKR, C3H/He, CBA, and ICR mice. The ovarian mast cells expressed connective tissue mast cell markers, although mast cells around the surface epithelium also expressed a mucosal mast cell marker in MRL/MpJ mice. Some ovarian mast cells migrated into the oocyte nests and directly contacted the compressed and degenerated oocytes. In MRL/MpJ mice, the number of oocytes in the nest was significantly lower than in the other strains, and the number of oocytes showed a positive correlation with the number of ovarian mast cells. The gene expression of a mast cell marker also correlated with the expression of an oocyte nest marker, suggesting a link between the appearance of ovarian ? 4mast cells and early follicular development. Furthermore, the expression of follicle developmental markers was significantly higher in MRL/MpJ mice than in C57BL/6 mice. These results indicate that the appearance of ovarian mast cells is a unique phenotype of neonatal MRL/MpJ mice, and that ovarian mast cells participate in early follicular development, especially nest breakdown.     

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。     

Presence of a blastocyst and mast cell depletion of the mouse uterus

The mast cell population has been studied during early decidualization of the mouse uterus. The number increases in early pregnancy until the attachment phase when a sharp depletion is noticed. This fall has been correlated with stimuli of maternal origin, but at the same time the presence of a blastocyst at the presumptive implantation sites seems to exert a significant effect on the depletion of mast cells. A relationship between the number of mast cells in the uterus and the physiological state of the organ has definitely been established [Harvey, 1964; Likar and Likar, 1964; Gibbons and Chang, 1972; Brandon and Evans, 1983]. The number of mast cells in the pregnant uterus is known to decrease around the time of implantation in the rat [Shelesnyak, 1960; De Feo, 1967; Brandon and Bibby, 1979]. Several workers believe that the mast cell population and histamine content decrease as a result of a rise in circulating estrogen [Westin, 1955; Gibbons and Chang, 1972; Spaziani, 1975]. In the present communication the possibility of a relationship between the decrease in mast cell population and the presence of a blastocyst, whose estrogenic role has been reported [Dickmann and Dey, 1973; Dickmann et al., 1975; Sengupta et al., 1977], in the uterine horn has been discussed.     

Compartmentalized mast cell degranulations in the ovarian hilum, fat pad, bursa and blood vessel regions of the cyclic hamster: relationships to ovarian histamine and blood flow.

Ovaries from hamsters on each day of the oestrous cycle at 09.00 h were observed for the number of mast cells, the pattern of mast cell degranulation, histamine concentration and blood flow. On day 4 (pro-oestrus), ovaries were also observed at 9.00, 15.00 and 21.00 h. Mast cell degranulation was evaluated by 3 criteria: (1) no degranulation = less than 5 granules dispersed from the cell; (2) moderate degranulation = 5 or more granules dispersed but less than 15, and (3) extensive degranulation = 15 or more granules released. Blood flow was determined using radio-active microspheres in anaesthetized animals. Mast cells were observed in fat pad (beyond 2 mm of the bursal mesothelium), bursa (within 2 mm of the bursal mesothelium), hilum and near ovarian blood vessels (these 4 regions are collectively called the ovarian complex). The distribution of ovarian mast cells was not uniform. Most mast cells were near ovarian blood vessels (42.2%) and in the fat pad (37.2%). A moderate number of cells were in the bursal wall (20%) and only a few cells were observed in the hilum (0.64%). Mast cell number remained unchanged on days 1-4 of the cycle in each ovarian compartment. However, summation of the number of mast cells in the entire ovarian complex revealed a significant decline in number at 15.00 h on pro-oestrus. Alterations in mast cell degranulation were primarily restricted to 2 periods of the cycle (pro-oestrus and di-oestrus). An increase in moderate but not extensive degranulation was observed in only the fat pad and bursa on day 2 when compared with day 1 values. In most ovarian compartments on pro-oestrus, degranulation was higher than on any other day of the cycle. At 15.00 h on pro-oestrus, extensive degranulation in bursa, fat pad and blood vessel regions (but not hilum) coincided with an increase in ovarian histamine and decline in number of mast cells; ovarian blood flow also increased at the time but remained unchanged the remainder of the cycle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Cause and effect relationship between myocardial mast cell number and matrix metalloproteinase activity

The objectives of this study were to investigate the temporal response of left ventricular (LV) matrix metalloproteinase (MMP) activity and collagen volume fraction (CVF) induced by an aortocaval fistula and the role of cardiac mast cells in regulating MMP activity. LV tissue was analyzed for MMP activity, CVF, and mast cell number in rats euthanized at 0.5, 1, 2, 3, 5, 14, 21, 35, and 56 days. Additional rats treated with the mast cell membrane-stabilizing drug cromolyn sodium were euthanized 1, 2, and 3 days postfistula. Marked increases in MMP activity occurred rapidly and remained significantly elevated for 5 days before returning toward normal. A significant decrease in CVF occurred by day 5, but thereafter CVF rebounded to normal or above normal values. The number of myocardial mast cells also significantly increased postfistula, and there was a close association between mast cell density and MMP activity. Cromolyn treatment prevented the increase in mast cell number and MMP activity. Thus it is concluded that cardiac mast cells play a major role in the regulation of MMP activity.     

Ontogeny of histamine-immunoreactive cells in rat stomach

Summary An antiserum against hemocyanin-conjugated histamine was used to study the cellular stores of histamine in the stomach, especially the oxyntic mucosa, of fetal and early postnatal rats. Tissues were fixed in 4% 1-ethyl-3(3-dimethyl-aminopropyl) carbodiimide (EDC-DI) and standard immunofluorescence technique was used. Histamine was first detected on the 16th embryonic (E16) day when a few histamine-immunoreactive (HA-ir) cells and nerve fibers were observed in the muscular layer of the stomach wall. On day E18, HA-ir cells were visualized for the first time in the oxyntic mucosa of the stomach, and from that day on the number of such cells increased slowly initially and after day E20 more rapidly. At birth many of the HA-ir cells in the oxyntic mucosa possessed processes giving them a paracrine-like appearance typical of enterochromaffin-like cells (ECL cells). Only a very small number of the HA-ir cells represented metachromatically stained mast cells and were located in the submucosa. After birth, the number of HA-ir ECL cells increased steadily, until day 21 when the distribution and number was very similar to that of the adult. The results suggest that histamine-containing neurons and ECL cells appear in the stomach wall before birth, and that there are histamine-containing ECL cells in the mucosa and mast cells in the submucosa of the stomach wall at birth.     

CD43 is relocated from the basal to the apical plasma membrane of rat uterine epithelial cells by progesterone

Adhesion molecules play an important part in preparing uterine epithelial cells for receptivity to the implanting embryo, and their rearrangement is crucial in allowing successful implantation. CD43 is an adhesion molecule which has previously been suggested to take part in implantation in mice. Indirect immunofluorescence microscopy localising CD43 was performed on uterine tissue during early pregnancy, and tissue obtained from ovariectomised rats administered with ovarian hormones. Western blotting was performed during early pregnancy on isolated epithelial cells and ovariectomised rats for comparison of the amount of CD43. Immunofluorescence microscopy showed CD43 was situated basally in uterine luminal epithelial cells on day 1 of pregnancy and during oestrogen administration, corresponding to a 95-kDa band of CD43 seen in western blotting. At the time of implantation, and during progesterone or progesterone plus oestrogen combined treatment, CD43 is apical in uterine luminal epithelial cells, resulting in an 85-kDa form of CD43. We suggest that a de-glycosylated form of CD43 moves from basally to apically at the time of implantation, thus facilitating blastocyst attachment to uterine epithelial cells as well as their removal.     

Mast cell changes in the organs of the immune system under physical loading

By means of histological and morphological methods reaction of mast cells has been studied in the thymus and inguinal lymph nodes of mature non-inbred white male rats, subjected to systematic physical loadings (daily swimming) with increasing time from 5 up to 100 min during 5 months. Morphological changes in the organs studied and manifestation of the mast cell reaction essentially depend on the degree of the animals' adaptation to the loading. In the animals adapted to swimming, decreasing area, occupied by the connective tissue elements, in comparison to that in the control--increasing cortical area, increasing number of lymphoid cells, decreasing number of the mast cells in the inguinal lymph nodes--are noted. When the adaptation of the animals to the loading is insufficient, outgrowth of the connective tissue elements, decreasing cortical zone, impoverishment of the parenchyma with lymphocytes occur. The number of the mast cells increases, many of them are at the state of degranulation.     

Bovine mast cells: distribution, density, heterogeneity, and influence of fixation techniques

Mast cells can be distinguished according to various characteristics: rodent mast cells have been subtyped by histochemical criteria, whereas canine and human mast cells have been classified according to their proteases. Comparisons of mast cells from different species have therefore resulted in contradictory and confusing opinions on mast cell heterogeneity. Thus, it is essential to obtain species-specific data on mast cell density and heterogeneity. The present study was carried out to determine the physiological distribution of mast cell numbers and types in bovines according to tissue location, staining, and fixation methods. Samples were fixed in formalin or Carnoy’s fluid. The average number of mast cells was determined by using a metachromatic staining method. Protease content of mast cells was examined with a double-enzyme-immunohistochemical staining technique. Three mast cell subtypes were distinguished: T-, TC-, and C-mast cells. The T-mast cell was the predominant subtype in nearly all investigated organs and tissue locations. Only tryptase-positive mast cells could be demonstrated in bovine skin and uterus. No chymase activity was found in these organs, regardless of the fixation type. A larger number of mast cells was observed after fixation in Carnoy’s fluid. The three different mast cell subtypes were only demonstrated in formalin-fixed tissue; chymase-positive mast cells were not found after fixation in Carnoy’s fluid. Increasing experimental data suggest that mast cell subtypes have different functions in promoting and modulating inflammation and in remodeling the extracellular matrix. Since mast cell tryptase and chymase have different functional properties, these results may clarify the different reaction patterns observed in various organs and species.     

Estrogen-induced increase in eosinophil number and peroxidase activity in uterus of genetically mast cell-deficient W/Wv mice

The role of mast cells in induction of uterine eosinophilia was investigated by using genetically mast cell-deficient (WB X C57BL/6)F1-W/Wv (hereafter called WBB6F1-W/Wv) mice. The injection of estradiol-17 beta (0.16 micrograms/g body weight) increased the peroxidase activity and eosinophil number in the uteri of castrated WBB6F1-W/Wv and WBB6F1-+/+ mice. Since no significant differences were detectable between these two type of mice, mast cells did not seem to be essential for the estrogen-induced uterine eosinophilia, at least in mice.     

Dihydrocapsaicin treatment depletes peptidergic nerve fibers of substance P and alters mast cell density in the respiratory tract of neonatal sheep

In the present study we administered dihydrocapsaicin (DHC) to neonatal lambs to deplete C-fibers of neuropeptides. We measured the density of substance P (SP)-fibers in nasal septum to assess the effectiveness of the treatment at 3, 9, and 21 days. The numbers of mast cells in the upper and lower respiratory tract were determined at the same time points and histamine content was determined from lung tissue. DHC treatment depleted SP-fibers for up to the 21 day time point. This depletion was estimated as 85% in comparison with controls. In vehicle-treated lambs, the density of SP-fibers decreased progressively with age, but not to the degree of DHC-treated lambs whose SP-fibers were depleted from the initial 3-day measurement. In both, vehicle- and DHC-treated lambs, numbers of mast cells increased progressively with time; however, the density of mast cells was augmented in the entire respiratory tract of DHC-treated animals. Apparently, DHC treatment exerts a single and initial effect in increasing mast cells whereas time maintains a continuous influence; both factors exert their influence independently. Despite large numbers of mast cells in DHC-treated animals, histamine content in the lung had similar levels as controls. Our study provides fundamental data for a better understanding of conditions that may influence defense mechanisms dependent on the mast cell-nerve axis in the respiratory tract.     

Heparin-binding EGF-like growth factor is seen on the extracellular surface of uterine epithelial cells only after the initial stages of blastocyst attachment

The presence and distribution of heparin-binding epidermal growth factor in rat uterine epithelial cells was determined immunohistochemically and localized ultrastructurally. Rat uterine tissue was examined on days 1, 3, 6 and 8 of pregnancy and it was found that while presence of this growth factor was evident from day 1, spatial reorganization occurred by the time of blastocyst implantation. Strong apical staining was evident from day 6 to day 8, day 6 being the approximate time of blastocyst implantation. Electron microscopy further revealed that this growth factor while shown to be expressed very strongly apically from day 6, actually localized on the plasma membrane only after attachment of the blastocyst. This suggests that heparin-binding epidermal growth factor is not involved in the initial stages of implantation but is more likely involved in the post attachment stages of pregnancy.     

Delayed mast cell mediated mitogenic reactivity in diabetic rats

Rats with diabetes of 4 weeks' duration have previously demonstrated increased mitogenesis in normal connective tissue cells following mast cell secretion. The appearance of this augmented mast cell-mediated mitogenic reactivity was studied in the mesentery of insulin-deficient rats, 3, 7, and 28 days after they had been rendered diabetic by a single dose of streptozotocin. On day 7 mast cell secretion induced a subnormal mitogenic response which, however, increased above normal on day 28. This time lag in the augmentation of mitogenic responsiveness may be important since proliferative lesions in diverse mesenchymal tissues typically develop with a similar delay in both experimental and clinical diabetes.