放线菌酮诱导大鼠脾细胞凋亡的电镜观察

利用透射电镜观察了放线菌酮体内诱导大鼠脾细胞凋亡的形态学变化，结果显示，腹腔注射放线菌酮４小时后，大鼠脾细胞发生凋亡，凋亡脾细胞核和胞质发生一系列形态学变化。主要表现为胞核染色质浓缩，重排，呈不同形态，多数凋亡脾细胞的粗面内质网增殖，凋亡小体形成，其线粒体也大量增加，或单个分布或团聚，肿胀，空泡化。结果提示，凋亡脾细胞的主要细胞器主动参与凋亡过程。     

放线菌酮诱导大鼠胸腺细胞凋亡的电镜观察

利用透射电镜详细观察了放线菌酮体内诱导大鼠腺细胞凋亡的形态学变化。观察显示，腹腔注射放线菌酮４小时后，大鼠胸腺细胞发生凋亡，凋亡胸腺细胞胞核和胞质发生一系列形态学变化，产生凋亡小体。主要表现为染色质断裂、浓缩、边集、大部分细胞核变成花瓣状，其它细胞核变成半月状、黑洞样和空泡状；粗面内质网大量增殖，并包裹细胞成分形成自噬体；线粒体增多，嵴紊乱并空泡化。凋亡细胞及其形成的凋亡小体被其它细胞吞噬清除。结     

放线菌酮诱导大鼠肠系膜淋巴结淋巴细胞凋亡

本实验利用放线菌酮建造细胞凋亡动物模型 ,用原位末端标记和电镜方法观察了大鼠肠系膜淋巴结淋巴细胞凋亡的形态学变化。结果显示放线菌酮处理 4h后 ,肠系膜淋巴结部分淋巴细胞处于凋亡状态 ,主要表现为细胞核凝聚 ,染色质边集 ,可见线粒体、内质网增殖 ,自噬体和凋亡小体形成。结果表明放线菌酮主要诱发肠系膜淋巴结生发中心的 B淋巴细胞凋亡 ,线粒体、内质网的增殖是细胞凋亡的一种主动行为。     

实验性糖尿病对大鼠胸腺皮质上皮细胞的影响

目的　探讨糖尿病对大鼠胸腺皮质上皮细胞影响的形态学变化 ,了解其引起胸腺细胞凋亡及胸腺萎缩的可能机制。　方法　采用四氧嘧啶型糖尿病动物模型 ,电镜下观察了糖尿病大鼠胸腺皮质上皮细胞的超微结构变化。　结果　 2型上皮细胞发生 4种变化 :1 内分泌功能紊乱样变 ,以大量变性的空泡与囊泡为特征 ;2 退化样变 ,以大量的变性分泌泡、髓样小体和粗大张力细丝束为特征 ;3 吞噬细胞样变 ,以大量的溶酶体样致密颗粒及吞噬的凋亡细胞与小体为特征 ;4 细胞凋亡样变 ,以细胞凋亡的早期形态学变化为特征。 3型上皮细胞发生两种变化 :1 退化样变 ,伴有大量分泌肽类激素样的致密颗粒群。 2 增生样变 ,在其胞体周伴有大量的胶原原纤维。此外 ,在胸腺皮质中还发现大量的胸腺细胞凋亡及胸腺细胞数量减少。　结论　糖尿病可导致大鼠胸腺皮质上皮细胞发生多样性的超微结构变化 ,可能是影响胸腺细胞发育 ,并导致其凋亡及胸腺萎缩的主要机制之一。     

放线菌酮诱导大鼠脾淋巴细胞凋亡的初步探讨

近来研究认为，细胞凋亡（Ａｐｏｐｔｏｓｉｓ）与免疫细胞分化发育有着密切关系。但目前，凋亡与淋巴细胞的关系的研究大多采用体外实验方法，为了更有利于研究淋巴细胞凋亡的发生机制，我们建立了体内诱导大鼠脾脏淋巴细胞凋亡的方法。健康雌性ＳＤ大鼠禁食２４小时后，用乙醚麻醉后从腹腔或静脉注射５．０ｍｇ，ｋｇ－１的放线菌酮（Ｃｙｃｌｏ－ｈｅｘｉｍｉｄｅ）。４小时后处死动物，取出脾脏，常规固定、脱水和石蜡包埋。细胞凋亡判断依据光镜下典型的凋亡细胞形态变化并结合原位３′－末端标记方法。脾脏组织还进一步作免疫组化染色…     

创伤后应激障碍大鼠杏仁核神经元凋亡及三偏磷酸酶活性的变化

目的:研究创伤后应激障碍(PTSD)大鼠杏仁核神经元细胞凋亡及三偏磷酸酶(TMP)活性的变化.方法:成年健康雄性Wistar大鼠50只,随机分为连续单一应激(SPS) 模型的1、 4、 7、 14 d组及正常对照组.应用Annexin V-FITC/PI双标记流式细胞技术定量检测及分析PTSD大鼠杏仁核神经元细胞凋亡率;利用透射电镜观察杏仁核神经元超微结构改变;利用酶组织化学方法检测杏仁核神经元TMP酶活性的变化.结果:SPS后,杏仁核神经元发生了凋亡的特征性形态学改变,杏仁核神经元细胞凋亡率显著增加,TMP酶活性明显比正常对照组增强,均于第4日达高峰.7 d和14 d凋亡率逐渐减少、TMP酶活性减弱.结论:PTSD大鼠杏仁核神经元发生细胞凋亡,凋亡率增加的同时TMP酶活性相应增强,提示TMP酶参与杏仁核神经元凋亡产物的吞噬.     

肠淋巴液引流减轻失血性休克大鼠脾组织损伤的作用与机制

 目的：观察肠淋巴液引流对失血性休克大鼠脾组织形态、细胞凋亡、细胞周期与增殖指数的影响,探讨肠淋巴液在休克发病学中的意义。方法: 在休克组与休克+引流组大鼠中复制失血性休克模型,低血压1.5 h后行液体复苏,休克+引流组大鼠于低血压1 h起引流休克肠淋巴液至液体复苏结束后3 h。留取固定位置的脾组织,HE染色观察脾组织形态,Hoechst 33258染色观察细胞凋亡,免疫组织化学染色检测Bcl-2和Bax蛋白表达,流式细胞术检测细胞周期和p53蛋白表达,计算增殖指数。结果：与假手术组比较,休克组大鼠脾组织出现了形态学损伤,凋亡细胞显著减少,Bax与p53蛋白表达显著升高,Bcl-2表达下降;休克+引流组大鼠脾组织G2/M期细胞数显著增多。与休克组比较,休克+引流组大鼠脾组织的损伤程度较轻,凋亡细胞和G0/G1期细胞显著减少,Bax与p53表达显著降低,G2/M期细胞显著增多,Bcl-2表达与增殖指数显著升高。结论: 休克肠淋巴液引流减轻了失血性休克大鼠的脾损伤,其作用机制可能与减少脾细胞凋亡有关。     

航天特因环境下抑郁模型大鼠海马形态学变化

目的研究航天特因环境下郁证大鼠模型海马形态的变化。方法采用经典抑郁模型,从海马形态学研究航天郁证,具体考察大鼠海马组织CA1、CA2、CA3、CA4区形态学变化。结果海马病理切片显示,大鼠海马CA1区锥体细胞偶见散在的胞体固缩,深染或胞质溶解、空泡变性。CA4区可见少量锥体细胞胞体固缩,深染。海马齿状回细胞未见明显病变。结论航天特因环境下郁证可能导致海马CA1和CA4神经元的改变。     

缺氧导致体外培养乳鼠心肌细胞凋亡的研究

目的:通过体外培养乳鼠心肌细胞模拟缺氧模型,探讨心肌细胞缺氧损伤时是否发生心肌细胞凋亡。方法:取(1~3)d新生SD乳鼠心肌进行心肌细胞培养,采用模拟缺氧模型,通过HE染色、流式细胞仪等手段观察检测细胞凋亡的改变。结果:在本实验心肌细胞缺氧模型中,HE染色可见凋亡细胞体积变小,胞核浓缩,胞质嗜酸性增强。流式细胞仪检测出明显的凋亡峰。结论:本实验培养乳鼠心肌细胞缺氧模型可导致心肌细胞凋亡,可通过HE染色观察凋亡细胞形态学的变化,流式细胞术检测细胞DNA含量而对凋亡细胞进行定量分析。     

鸡胚脾脏凋亡细胞核及其环孔板的超微结构

目的：着重观察鸡胚脾脏凋亡细胞核及其环孔板的形态学变化。方法,用放线菌酮处理第１５天胚龄鸡胚,建立鸡胚脾脏细胞凋亡模型,用透射电镜进行观察。结果：凋亡细胞为各类幼稚血细胞,以幼稚淋巴细胞为主,巨噬细胞未见凋亡。凋亡细胞核在裂解分离为核碎块的过程中出现环孔板,凋亡细胞核完全裂解分离后,环孔板消失。结果：环孔板参与凋亡细胞核的分割以及核碎块的包囊,提示凋亡细胞核裂解成为核碎块形凋记录上体的过程是一个主     

Morphine Induces Splenocyte Apoptosis and Enhanced mRNA Expression of Cathepsin-B

Morphine has been demonstrated to modulate immune function. We studied whether morphine modulates apoptosis of splenocytes. Splenocytes were isolated from control and morphine treated rats. Splenocytes isolated from morphine treated rats showed increased percentage (P < 0.001) of apoptosis when compared to splenocytes isolated from untreated rats (control, 4.7 ± 1.0% apoptotic splenocytes/field vs. morphine, 47.8 ± 3.4% apoptotic splenocytes/field). These results were further confirmed by gel electrophoresis as well as by end-labeling DNA of splenocytes isolated from control and morphine treated rats. Splenocytes from morphine treated rats showed a classical ladder pattern with integer multiples of 180 base pairs. Splenocytes from morphine treated rats also showed increased mRNA expression of cathepsin-B, a gene associated with active cell death. These results suggest that morphine may also be modulating immune function by enhancing apoptosis of splenocytes.     

The relationship between apoptosis and splenocyte depletion in rats following ethanol treatment

Splenocyte depletion observed in chronic ethanol-treated rats (ETRs) was studied in relation to apoptosis. The rats were fed with ethanol in a Liber-DeCarli liquid diet (36% of total calories as ethanol) for 7 weeks. Spleens of ETRs and control rats were examined by the terminal deoxynucleotidyl transferase-mediated dUTP nick end-labeling (TUNEL) method, immunohistochemistry using anti-rat p53 and RM4 (specific for macrophages) monoclonal antibodies, and transmission electron microscopy (TEM). The splenic white pulp in ETRs decreased in size and showed a moth-eaten appearance because of the severe depletion of splenocytes. Most TUNEL-positive cells aggregated into clusters or nests and were not isolated in the white pulp of ETRs. The site of RM4 immunoreactivity was consistent with that of clusters of TUNEL-positive cells. The p53 immunoreactivity was observed in apoptotic splenocytes that were isolated or phagocytosed by macrophages. TEM study revealed the increase in tingible body macrophages phagocytosing apoptotic splenocytes in their cytoplasm in ETRs. Chronic ethanol intake certainly induces apoptosis in splenic white pulps, and tingible body macrophages act as both sentinels and scavengers of apoptotic splenocytes expressing p53.     

孕激素对更年期雌性大鼠脾细胞凋亡及相关基因的调控作用

为了解孕激素对更年期雌性大鼠脾细胞凋亡及相关基因的调控作用。首先通过应用Annexin V-FITC/PI双染色流式细胞仪测定脾细胞凋亡率;通过RT-PCR检测脾细胞凋亡基因Bcl-2、Bax的表达;RIA法测血孕酮(P)水平。结果发现更年期组血孕激素水平明显低于青年组和孕激素组(P<0.01),孕激素组与青年组相比差别无显著意义(P>0.05);更年期组大鼠脾细胞凋亡率高于青年组(10.3%和7.7%,P<0.05),促凋亡基因Bax的转录水平也高于青年组;孕激素组脾细胞的凋亡率与更年期组比较无显著差异,孕激素组促凋亡的Bax基因转录水平高于更年期组大鼠(P<0.01),而Bcl-2基因未见显著差异。研究结果提示,衰老过程中脾淋巴细胞凋亡增加,并受到相关基因的调控,孕激素治疗未能逆转更年期雌性大鼠脾细胞凋亡的增加,且使更年期大鼠脾细胞促凋亡的Bax基因表达上调。     

Mechanism of perfluorooctanesulfonate (PFOS)-induced apoptosis in the immunocyte

As a new type of persistent organic pollutant, perfluorooctane sulfonate (PFOS) has raised great concern in recent years due to its ubiquitous distribution in the general environment and its long elimination half-life in humans. PFOS has toxic and carcinogenic effects in animals and humans, but the effects of PFOS on apoptosis are still not clear. The present study aimed to determine the mode of cell death and its mechanism in splenocytes and thymocytes from adult male C57BL/6 mice administered 0, 1, 5, or 10 mg PFOS/kg/day by gavage daily for 7 days. The results showed that more apoptotic cells were present in PFOS-treated mice than in control mice. PFOS induced production of reactive oxygen species (ROS), dissipation of mitochondria membrane potential, and apoptosis of splenocytes and thymocytes. Moreover, activities of superoxide dismutase, catalase, and glutathione reductase were increased, whereas activities of glutathione-S-transferase and glutathione peroxidase were decreased, in splenocytes. Glutathione contents were reduced as well. Differential expressions of proteins such as p53, Bax, caspase-3, and caspase-9 were significantly up-regulated in PFOS-exposed hosts, whereas Bcl-2 expression was significantly down-regulated. One possible mechanism for the findings here was that PFOS could overwhelm homeostasis of anti-oxidative systems, boost ROS generation, impact on mitochondria, and affect protein expression of apoptotic regulators, the latter of which resulted in initiation of the apoptosis program. Results from this study may provide a new insight into the potential adverse effects of PFOS exposure on humans, at the cellular level.     

植物雌激素对去卵巢大鼠海马神经元线粒体超微结构的保护作用

采用去卵巢(OVX)SD大鼠模拟女性绝经后神经元的退变来观察雌激素、植物雌激素对海马神经元线粒体的保护作用。雌性SD大鼠分为5组:(1)正常对照组;(2)去卵巢对照组;(3)雌激素治疗组;(4)金雀异黄酮治疗组;(5)依普拉芬治疗组,并于术后12d取材进行超微结构观察和体视学定量分析。结果显示:(1)去卵巢后海马神经元的线粒体肿胀明显,嵴断裂明显,空泡化显著;3个治疗组细胞和线粒体相对于去卵巢对照组均有明显改善;(2)与去卵巢对照组相比较,3个治疗组线粒体的体积密度(Vv)、平均直径(D)、平均表面积(S)明显减小(P<0.05);3个治疗组的比表面(δ)、数密度(Nv)和粒子分散度(Cλz)较去卵巢对照组明显增大(P<0.05)。以上结果表明:雌激素和植物雌激素对去卵巢后大鼠的海马神经元线粒体具有保护作用。本研究结果为雌激素和植物雌激素以线粒体为靶点防治女性更年期后神经元的退变及老年性痴呆可能的作用机制提供了一定的形态学资料。     

老年学习记忆减退大鼠海马GABA能神经元树突内线粒体的变化

为进一步探讨ＧＡＢＡ 能神经元与老年学习记忆减退的关系，本研究以老年学习记忆减退大鼠为模型，用免疫电镜结合体视学方法观察了老年学习记忆减退大鼠海马ＣＡ１ 区放射 分子层的ＧＡＢＡ 能神经元树突内线粒体的改变。结果显示，线粒体的面数密度在老年学习记忆损害组明显小于青年组和老年正常组（Ｐ＜ ０．０１）；体密度的绝对值有所减少，但无统计学意义（Ｐ＞０．０５）。此外，衰老时较小的树突内线粒体的密度增大，老年学习记忆正常组小直径树突内线粒体面数密度的增加较老年学习记忆损害组更明显。结果提示，衰老过程中，ＧＡＢＡ 阳性树突内线粒体的数目减少，单个线粒体的体积代偿性增大。老年学习记忆减退时，树突结构和功能的内源性代偿能力下降，可能处于一种失代偿状态。     

Swelling of free-radical-induced megamitochondria causes apoptosis

Recently, we have found that cultured cells from various sources exposed to free radicals become apoptotic in the presence of megamitochondria (MG). The purpose of the present study is to answer the following two questions: (1) Do functions obtained from the "MG fraction" isolated from normal mitochondria by a routine procedure represent the functions of MG since the fraction consists of enlarged and normal-size mitochondria? (2) What is the correlation between MG formation and apoptotic changes of the cell? In the present study the heavy fraction rich in mitochondria enlarged to varying degrees and the light fraction consisting mainly of normal-size mitochondria were isolated independently from the livers of rats treated with hydrazine for 4 days (4H animals) and 8 days (8H animals), and some functions related to apoptosis were compared. Results were as follows: (1) Mitochondria in both fractions obtained from 8H animals swelled far less in various media than those obtained from the controls, suggesting that the permeability transition pores had been opened before they were exposed to swelling media. (2) The membrane potential of mitochondria in both fractions obtained from 8H animals was distinctly decreased. (3) The rates of reactive oxygen species generation from mitochondria of both fractions in 4H animals were equally elevated, while those in 8H animals were equally decreased compared to those of controls. These results, together with morphological data obtained in the present study, suggest that enlarged and normal-size mitochondria are a part of MG and that the secondary swelling of MG causes the apoptotic changes in the cell.