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。     

上皮组织增生与肥大细胞变化的形态学研究

Ｃ－１号小鼠４０只,随机分为４组,每组１０只,用食醋抹小鼠右耳皮肤,每日一次,以左耳作对照。在实验后第１０、３０、９０、１４０天各处死一组,取耳廓组织,常规石蜡制片,进行ＨＥ染色及肥大细胞染色,光镜下观察。结果发现：实验组１４０天皮肤上皮组织明显的增生,结缔组织内肥大细胞（ＭＣ）的数量比对照组显著增多（Ｐ＜０．０１）。在ＡＢ－Ｓ染色时,实验组ＭＣ由藏红阳性为主逐渐转变成以Ａｌｃｉａｎ蓝阳性为主,而对照组各时期ＭＣ均呈藏红阳性。实验组与对照组皮肤ＭＣ的临界电解质浓度分别为０．９５Ｍ,０．９７Ｍ。硫酸小蘖碱荧光染色均呈强黄色萤光反应。结果提示：①上皮组织增生可致结缔组织内ＭＣ数量增多,②增多的ＭＣ的组化性质发生了变化,③ＭＣ可参与上皮组织的增殖,ＭＣ与上皮细胞间具有相互促进作用     

改良的肥大细胞甲苯胺蓝染色法

目的:本研究旨在运用一种改良的甲苯胺蓝染色方法鉴定体外分离培养的肥大细胞形态,为肥大细胞研究提供简便的检测方法。方法:体外诱导分化培养小鼠骨髓来源的肥大细胞,4周后收集细胞、固定、染色。比较不同固定温度及时间对甲苯胺蓝染色效果的影响,确定最佳条件。结果:SCF和IL-3体外培养诱导出小鼠骨髓来源肥大细胞。肥大细胞用改良的甲苯胺蓝染色后细胞着色效果较好,细胞多呈圆形或椭圆形且胞膜较完整,胞浆中充满大量的紫红色颗粒。结论:本研究成功运用一种适用于体外培养小鼠骨髓源肥大细胞的甲苯胺蓝染色法,并发现肥大细胞在37℃充分固定后进行染色,可以降低肥大细胞发生脱颗粒。此操作方法稳定性好、简便,适用于体外培养的肥大细胞形态学观察。     

从甲状腺肥大细胞探讨胎儿器官肥大细胞的差异

研究肥大细胞在人胎儿甲状腺发育中数量、分布及组化性质的改变,以探讨胎儿器官发育中肥大细胞的差异。取45例不同胎龄的人胎甲状腺石蜡切片做甲苯胺蓝染色和阿尔辛蓝－－藏红染色,并测定肥大细胞的临界电解质浓度值及进行硫酸小蘖硷荧光染色。结果显示：3月龄胎儿甲状腺内开始出现肥大细胞,数量极少,主要分布在被膜及小叶间结缔组织内,甲苯胺蓝染色肥大细胞颗粒呈淡紫蓝色,阿尔辛蓝－－藏红染色呈蓝色,临界电解质浓度值较低,硫酸小蘖硷染色未见显黄色荧乐的肥大细胞,从3月龄到足月随着胎龄增长,肥大细胞数量缓慢增多,8月龄时肥大细胞经甲苯胺蓝染色,其颗粒呈紫红色,阿尔辛蓝－－藏红染色出现少量含红色和红蓝混合染色颗粒的肥大细胞,临界电解质浓度值偏高,可见少量显黄色荧光的肥大细胞,结果表明：在人胎儿3月龄时甲状腺发育中开始出现肥大细胞,但随胎儿发育肥大细胞的组化性质改变不明显。     

不同固定液和染色方法显示山羊子宫内肥大细胞效果的比较

目的探讨用不同固定液和染色方法对显示处于间情期山羊子宫肥大细胞的影响。方法用四种不同的固定方法,应用改良甲苯胺蓝（MTB）染色法和阿尔辛蓝-番红花红（AB-S）染色法显示处于间情期山羊子宫肥大细胞。结果山羊子宫组织采用Carnoy氏液固定,MTB和AB-S染色对所有的肥大细胞均可获得良好的染色反应,但10％中性福尔马林,4％多聚甲醛,Bouin氏液固定的组织仅有少量肥大细胞着染。结论MTB和AB-S染色法均是山羊子宫肥大细胞良好的染色方法。     

食管癌胃癌间质中肥大细胞淋巴细胞光镜与电镜研究

对３５例食管癌与３０例胃癌间质中的肥大细胞与淋巴细胞进行光镜与电镜观察。结果发现,两种癌间质中的肥大细胞数量在恶性程度低组均比恶性程度高组与对照组显著增加。在各组食管癌或胃癌中,有淋巴细胞高度浸润并形成淋巴小结者,肥大细胞也增多。用Ａｌｃｉａｎ蓝—藏红染色,对照组Ａｌｃｉａｎ蓝阳性和藏红阳性的肥大细胞都可以见到;而在食管癌胃癌间质中,大部分肥大细胞均呈Ａｌｃｉａｎ蓝阳性。电镜观察,对照组肥大细胞颗粒多具有涡卷状结构,而癌组者则显示多样化形态。一些肥大细胞和淋巴细胞与癌细胞接触,肥大细胞可见活跃脱颗粒现象。结果提示,在抗肿瘤生长的防御机制中,肥大细胞与淋巴细胞有协同作用。     

动情周期中大鼠子宫和输卵管壁肥大细胞数量变化的研究

用放射免疫分析法对动情周期中大鼠血清雌二醇浓度进行检测;取子宫、输卵管常规石蜡切片、H－E染色,并用甲苯胺蓝染色显示肥大细胞,于光镜低倍视野下计数。结果显示：动物血清雌二醇浓度依次为：动情期（E）组＞动情前期（PE）组＞动情后期（ME）组＞动情间(DE)且,各组间差异均有显性;在子宫,肥大细胞分布于宫壁肌怪平滑肌束间的结缔组织内、近小血管处,以微血管周居多,常见单个散在,于ME子宫内膜尚偶见肥大细胞;输卵管肥大细胞局限于其外膜层内、近小血管周围,亦多散在。子宫、输卵管壁内的肥大细胞镜下呈圆形、椭圆形或略不规则形,胞浆内充满紫红色粗大颗粒,子宫肥大细胞数量依次为：ME＞DE＞PE＞E,各组间差异有生（DE与PE、PE与E,P＜0.05,余组间P＜0.01）;输卵管壁内肥大细胞数量各组间差异无显性（P＞0.05）。本尚对大鼠血清雌二醇水平波动与子宫、输卵管壁内肥大细胞数量变化的关系及其生理意义进行了讨论。     

奥尼罗非鱼胃肠道中肥大细胞的分布特征

本研究采用改良甲苯胺蓝染色法探讨了奥尼罗非鱼(Oreochromis niloticus ♀×O. aureus♂)胃肠道肥大细胞的分布及其形态特点。结果发现,经甲苯胺蓝染色的肥大细胞其核着深蓝色,颗粒被染成紫红色,着色深浅不一。肥大细胞大小不一,形态各异,呈圆形、椭圆形或梭形、菱形,散在或集中分布在黏膜层固有膜和黏膜下层,尤其常见分布于小血管周围。经统计,肥大细胞在奥尼罗非鱼的胃、幽门盲囊、后肠、前肠、中肠的数量依次减少。胃和幽门盲囊内肥大细胞数量显著高于前肠和中肠(P0.05),与后肠无显著差异;前肠、中肠和后肠内肥大细胞数量并无明显差别。     

常用实验动物不同组织部位肥大细胞异质性的组织学特点比较

目的探讨并比较六种常用实验动物不同部位肥大细胞异质性的形态学特点。方法应用麻醉后处死的方法,取小鼠、大鼠、豚鼠、兔、犬和猴的皮肤、肺脏、乙状结肠和脾脏组织,经4%中性缓冲甲醛溶液或Bouin’s液固定后,制作常规组织切片,分别作HE、甲苯胺蓝、阿尔新蓝-藏红和P物质免疫组织化学染色,镜下观察并应用彩色病理图像分析软件进行形态学分析;另取皮肤组织固定于磷酸缓冲戊二醛溶液,制作常规超薄切片,透射电镜下观察肥大细胞超微结构。结果六种动物不同组织中肥大细胞各有其分布特点,且在形态大小、异染性及染色特性等方面表现各异,肥大细胞密度和形态学参数差异有显著性（P〈0.05）;豚鼠、犬和猴皮肤肥大细胞颗粒具有特殊亚微结构;小鼠皮肤组织内可见P物质免疫阳性肥大细胞和神经纤维,犬脾脏内可见P物质免疫阳性肥大细胞。结论在六种实验动物的同一组织中以及同一种动物不同组织中,肥大细胞具有明显的异质性,此异质性对采用实验动物开展与肥大细胞功能相关的动物实验研究具有重要参考价值。     

肥大细胞用甲苯胺蓝染色之体会

肥大细胞是存在于多种组织内的一类细胞 ,并因所在组织器官的不同而具有不同的功能。肥大细胞是抗感染免疫的第一线细胞 ,在疾病过程中具有重要作用 ,因此 ,研究肥大细胞的生物学性质具有重要的理论意义和广泛的应用价值。常用的肥大细胞染色方法有甲苯胺蓝染色法、阿尔辛蓝—番红染色法等。肥大细胞具有复杂的异质性 ,因此上述方法不仅难以全面反映其生物学特性 ,而且有时也难客观地反映其在组织中的分布。我们用改良的甲苯胺蓝染色法对食管、肠、子宫、乳腺等器官中的肥大细胞进行染色时得到了一点体会 ,供参考。甲苯胺蓝改良染色法 :根据…     

An assessment of mast-cell deficient mice (W/Wv) as a model system to study the role of histamine in implantation and deciduoma formation

The ovaries from mast cell-normal (+/+) and mast cell-deficient (W/Wv) mice were examined with light and electron microscopy. In addition the effect of ovariectomy and subsequent steroid treatment on total uterine histamine content, total mast cell numbers and surface and glandular epithelial cell heights was measured. The ovaries of +/+ mice were normal, displaying various stages of follicular growth and atresia and numerous corpora lutea; the ovaries of W/Wv mice lacked follicles and corpora lutea but contained numerous hyperplastic interstitial cells which contained numerous lipid droplets, vesiculated mitochondria and abundant endoplasmic reticulum suggestive of steroid synthesis. Steroid treatment of ovariectomized +/+ and W/Wv mice caused a significant increase in uterine wet weight and endometrial surface and glandular epithelial cell heights. In +/+ mice, steroid treatment caused a concomitant increase in total mast cells per uterine horn while mast cells were totally absent in W/Wv mice. The increase in uterine histamine in +/+ mice is consistent with the increase in mast cell numbers. Measurable amounts of uterine histamine, which increases slightly after steroid treatment, were demonstrated in W/Wv mice. Since the uteri of +/+ and W/Wv mice respond to steroids in a similar manner with the sole exception being histamine content and mast cell numbers, our results demonstrate the potential of using these animals to investigate the role(s) of uterine mast cells and non-mast cell uterine histamine in the process of implantation and the formation of a decidual cell response.     

舒必利与垂体移植诱发小鼠子宫腺肌病的比较

目的比较舒必利诱导和垂体移植诱导两种造模方法的效果,评价适用于子宫腺肌病研究的小鼠模型。方法 45只7周龄未曾受孕的雌性BALB/c小鼠,随机分为对照组、舒必利组和垂体移植组,每组15只。舒必利组每日皮下注射舒必利,每20 g体重注射800μg,垂体移植组则将同种同龄雄鼠的垂体放入雌鼠右侧子宫内,对照组不予任何处理。5个月后以子宫湿重、终末体重、子宫湿重/终末体重的比值、卵巢湿重、子宫HE染色评分及模型成功率等指标评价造模效果。结果 5个月后舒必利组和垂体移植组小鼠的子宫湿重、子宫湿重/终末体重的比值、子宫HE染色评分及成功率均显著高于对照组,但两模型组各项指标比较差别无统计学意义,三组小鼠终末体重和卵巢湿重比较差别均无统计学意义。结论对照组无一例形成子宫腺肌病,舒必利诱导和垂体移植诱导两种造模方法均可引起小鼠子宫腺肌病,造模的各项评价指标均无显著差异,都可用于子宫腺肌病的研究。     

Inhibitory effect of progesterone on cell death of mouse uterine epithelium

The protective effect of progesterone against cell death of mouse uterine epithelium was evaluated by examining the retention of 5'-[125I]iodo-2'-deoxyuridine [( 125I]IdUrd) incorporated into the whole uterus and the apoptotic index (percentage of apoptotic cells in total cells), which is a good index of physiological cell death. Castrated adult female mice were given a daily injection of oestradiol-17 beta for 3 days, and then an injection of [125I]IdUrd. They were then divided into 4 groups, which received a daily injection of vehicle only, oestradiol-17 beta (E), progesterone (P), or both oestradiol-17 beta and progesterone (EP), and were killed at intervals during these treatments for determination of 125I radioactivity retained in the whole uterus. On treatment with vehicle only, the 125I radioactivity retained in the uterus decreased rapidly, but treatment with E, P or EP reduced the loss of 125I radioactivity significantly. Progesterone did not antagonize the effect of oestradiol-17 beta on the 125I radioactivity retained in the uterus. The apoptotic index of uterine cells was examined by a similar experimental protocol, but without injection of [125I]IdUrd. In the group treated with vehicle only, the apoptotic indices of both luminal and glandular epithelia increased markedly, but the injection of E, P or EP suppressed these increases significantly. Progesterone did not antagonize the effect of oestradiol-17 beta on the apoptotic index. The apoptotic index of stroma was not affected by the injection of E, P or EP. On the other hand, progesterone completely inhibited the increase in the mitotic index of uterine epithelia induced by oestradiol-17 beta. These results show that progesterone alone or in combination with oestrogen reduced cell death in mouse uterine epithelium and that the effects of oestrogen and progesterone on uterine cell death were independent of their actions on cell division.     

The influence of ovarian hormones on the rat oviductal and uterine concentration of noradrenaline and 5-hydroxytryptamine

This paper describes the effects of estradiol and progesterone on the concenirations of noradrenaline and 5-hydroxytryptamine in the Wistar rat oviduct and uterus. The levels of noradrenaline and 5-hydroxytryptamine are higher in the oviduct than in the uterus whereas p-tyrosine and tryptophan are similar in both tissues. Estradiol treatment reduced the oviductal concentration of noradrenaline but not 5-hydroxytryptamine in oviduct, while the concentrations of both noradrenaline and 5-hydroxytryptamine were reduced in uterine horn. The levels of noradrenaline in the oviduct and uterus in rats in estrus were lower than those of diestrous rats. Bilateral ovariectomy produced an increase in uterine noradrenaline and 5-hydroxytryptamine levels. These changes were reversed in the presence of ovarian hormones as indicated by experiments where unilateral ovariectomy was performed. Reserpine administration reduced noradrenaline concentration in both the oviduct and the uterus but did not change oviductal or uterine 5-hydroxytryptamine.These results indicate the existence of noradrenaline within postganglionic sympathetic nerve terminals and suggest that estrogens increase the utilization and the synthesis of noradrenaline in both the oviducts and the uterine horns. With respect to 5-hydroxytryptamine the data support the concept that it is mainly associated with mast cells.     

Effects of Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae) larvae on the degranulation of dermal mast cells in mice; an electron microscopic study

The pine caterpillar Thaumetopoea pityocampa (Lepidoptera: Thaumetopoeidae) is found in pine woods. Hairs of the T. pityocampa caterpillar cause a cutaneous reaction in humans and animals. Mast cells are responsible for allergic reactions in mammals. In this study male swiss albino mice were divided into two groups: 5 mice in the control group and 25 mice in the experimental group. The dorsal skin of mice was shaved. The mice in the experimental group and T. pityocampa larvae (fifth instar, approximately n=100) were put in the same cage. Dermal mast cells of mice exposed to T. pityocampa were examined with a transmission electron microscope and compared to the control group 1, 3, 6, 12 and 24 hours after exposure. Dermal mast cell degranulation in mice was observed 12 and 24 hours after exposure.     

Inhibitory effect of androgen on cell death of mouse uterine epithelium

The protective effect of androgen against the cell death of mouse uterine epithelium was evaluated by examining the retention of 5'-[125I]iodo-2'-deoxyuridine ([125I]IdUrd) incorporated into the whole uterus and the apoptotic index (percentage of the apoptotic cells to the total cells) which is a good index of physiological cell death. Castrated adult female mice were daily injected with oestradiol-17 beta for 3 days, followed by the injection of [125I]IdUrd. Thereafter, these mice were daily injected with only the vehicle or 5 alpha-dihydrotestosterone (DHT), and the 125I-radioactivity retained in the whole uterus was determined. When only the vehicle was injected, the 125I-radioactivity retained in the whole uterus rapidly decreased but injections of DHT reduced the loss of 125I-radioactivity. The effect of DHT on the retention of 125I-radioactivity depended on doses of DHT and was abolished by the pure antiandrogen, flutamide. The apoptotic index of uterine cells was examined by a similar experimental protocol, but without an injection of [125I]IdUrd. Injections of only the vehicle caused marked increases in the apoptotic indices of both luminal and glandular epithelia, but injections of DHT decreased them significantly. The apoptotic index of stroma was not affected by the injection of DHT. The present results indicated that androgen reduces the cell death of mouse uterine epithelium through the androgen receptor.     

Uterine contractions depend on KIT-positive interstitial cells in the mouse: genetic and pharmacological evidence

In the gastrointestinal tract, interstitial cells of Cajal (ICCs) generate a pacemaker activity. They produce electric slow waves that trigger and coordinate gut smooth muscle contractions. Interstitial cells of Cajal's slender shape is revealed by KIT immunostaining. Based on several features, including KIT expression and KIT dependence, ICC-like cells were identified in nongastrointestinal tissues. Here, we investigated in the mouse whether uterine contractions depend on ICC-like cells' activity. By labeling KIT-expressing cells, we found putative ICC-like cells in the uterus, observed as KIT-positive interstitial, long spindle-shaped cells with fine branched cytoplasm processes, distributed in muscular layers and in subepithelial connective tissue. We then checked the potential KIT dependence of ex vivo contractile activity of the uterus by combining genetic and pharmacological approaches, using the Kit W-v hypomorphic mutation, and imatinib as a KIT noncompetitive inhibitor. We found a significant reduction in frequency of longitudinal uterine contractions in Kit W-v/Kit W-v compared with Kit+/+ mice, whereas amplitude was unaffected. There was no difference in frequency or amplitude of circular uterine contractions between Kit W-v/Kit W-v and Kit+/+ mice. Ex vivo treatment of Kit+/+ uterine horns with imatinib resulted in a dose-dependent reduction of the frequency and amplitude of longitudinal myometrial contractions. Amplitude and frequency of circular contractions were unaffected in presence of imatinib. These concurrent results suggest that longitudinal contractions of the uterus depend on a KIT signaling pathway of ICC-like cells. The existence of ICC-like cells in the myometrium may enhance our understanding of uterine spontaneous contractile activity and suggest new approaches for treatment of uterine contractility disorders.