NGF对小脑皮质细胞凋亡影响的实验研究

目的　探讨神经生长因子 (NGF)对小脑皮质神经细胞凋亡的影响 ,为临床治疗小脑变性疾病提供新的措施。方法　以原代培养的新生SD乳鼠小脑皮质细胞建立谷氨酸诱导的神经细胞凋亡模型。为观察NGF对受损细胞的保护作用 ,应用MTT法测定细胞的存活率和细胞代谢情况 ;利用光学显微镜技术观察细胞凋亡的形态学改变。结果　Glu (5 0 0 μmol/L)作用 5min可诱导小脑神经细胞凋亡。MTT法计数结果显示 ,NGF高剂量治疗组细胞存活率显著高于模型组 (P <0 0 1) ,光学显微镜观察发现受损细胞的形态学变化也得到明显改善。结论　外源性NGF能够减轻大鼠小脑皮质神经细胞凋亡。     

Trehalose protects against ocular surface disorders in experimental murine dry eye through suppression of apoptosis

The disaccharide trehalose is a key element involved in anhydrobiosis (the capability of surviving almost complete dehydration) in many organisms. Its presence also confers resistance to desiccation and high osmolarity in bacterial and human cells by protecting proteins and membranes from denaturation. The present study used a novel murine dry eye model induced by controlled low-humidity air velocity to determine whether topically applied trehalose could heal ocular surface epithelial disorders caused by ocular surface desiccation. In addition, the efficacy of 87.6 mM trehalose eyedrops was compared with that of 20% serum, the efficacy of which has been well documented. Mice ocular surface epithelial disorders were induced by exposure of murine eyes to continuous controlled low-humidity air velocity in an intelligently controlled environmental system (ICES) for 21 days, which accelerated the tear evaporation. The mice were then randomized into three groups: the control group received PBS (0.01M) treatment; a second group received 87.6 mM trehalose eyedrops treatment; and the third group received mice serum eyedrops treatment. Each treatment was administered as a 10 μl dose every 6 h for 14 days. The resultant changes in corneal barrier function and histopathologic examination of cornea and conjunctiva were analyzed and the level of apoptosis on the ocular surface was assessed using active caspase-3. After 14 days of treatment, the corneal fluorescein staining area, the ruffling and desquamating cells on the apical corneal epithelium, as well as the apoptotic cells on ocular surface epithelium had significantly reduced in eyes treated with trehalose compared with those treated with serum and PBS. In contrast, after 14 days of treatment, improvements in the thickness of the corneal epithelium, the squamous metaplasia in conjunctival epithelium and the number of goblet cells of the conjunctiva were less marked in eyes treated with trehalose compared with serum. These results demonstrated that trehalose could improve the appearance of ocular surface epithelial disorders due to desiccation through suppression of apoptosis. Trehalose produces some of the same responses as serum upon topical application and can maintain corneal health.     

亚低温对大鼠短暂性脑缺血迟发性神经元死亡的影响

目的 观察亚低温（33℃）对大鼠短暂性脑缺血后神经元的保护作用。方法 32只DS大鼠分为假手术组、常温缺血组和即刻亚低温组，采用尼氏体亚甲蓝特殊染色观察存活神经元、原位细胞凋亡检测法（TUNEL染色）检测及电镜观察脑缺血后大鼠CA1区神经元凋亡情况。结果 与假手术组相比，常温缺血组海马CA1区存活的锥体细胞数目减少（P＜0．01）；与常温缺血组相比，亚低温缺血组海马CA1区存活的锥体细胞数目明显增多（P＜0．01）。亚低温缺血组大鼠海马CA1区TUNEL染色阳性细胞数目明显少于常温缺血组（P＜0．01）。结论 脑缺血后迟发性神经元死亡很可能通过凋亡途径，亚低温对缺血后神经元的保护作用与减少神经元凋亡有关。     

重组人白细胞介素6对SW872脂肪细胞增殖和凋亡的影响

目的　探讨白细胞介素 6(IL 6)对SW872脂肪细胞增殖及凋亡的影响。方法　体外培养SW872脂肪细胞 ,0 .6mmol/L油酸诱导SW872前脂肪细胞分化 ,化学比色法检测细胞内三酰甘油的总量 ;油红O染色观察细胞内脂肪的聚积 ;MTT法检测不同浓度IL 6对SW872前脂肪细胞增殖活力的影响 ;流式细胞仪分析IL 6对SW872成熟脂肪细胞凋亡的影响。结果　 0 .6mmol/L油酸刺激 72h ,几乎 10 0 %SW872前脂肪细胞分化为成熟的脂肪细胞 ,细胞内三酰甘油的含量明显增加 ,油红O染色胞浆内可见丰富的脂滴 ;5 μg/LIL 6对SW872前脂肪细胞的增殖没有影响 ,10～ 5 0 μg/LIL 6明显抑制SW872前脂肪细胞的增殖 ;IL 6对SW872成熟脂肪细胞的凋亡无影响。结论　IL 6能明显抑制SW872前脂肪细胞的增殖 ,其抑制效应呈剂量依赖性 ,表明IL 6可能与肥胖有关 ,高剂量IL 6可降低肥胖的程度。     

灯盏细辛黄酮类成分对压力诱导的视网膜神经节细胞凋亡的影响

目的考察灯盏细辛中黄酮类成分(总黄酮、灯盏花素)对体外高压诱导的视网膜神经节细胞(retinal gan-glion cells,RGCs)凋亡的影响,探讨其视神经保护作用的物质基础。方法采用胰酶消化法将24只出生2~3d的SD(Sprague-Dawley)乳鼠视网膜制成细胞悬液,接种于经多聚鸟氨酸(HA)和层粘连蛋白(LN)包被的盖玻片中。培养72h后,将覆有细胞的血盖片转入加压装置中,加入黄酮类成分,继续培养48h,采用Fas蛋白免疫组化染色法及原位末端脱氧核苷酸转移酶标记(Tunel-POD)法进行检测,每天观察细胞形态,同时对部分细胞行NSE染色检查。结果细胞生长良好,NSE染色表明,85%以上的细胞为RGCs。给药组的Fas蛋白阳性表达指数和凋亡指数均明显低于模型组(P<0.05~0.01)。结论黄酮类成分均能对抗压力诱导的RGCs凋亡,为灯盏细辛视神经保护的有效组分。     

脑内缝隙连接

缝隙连接普遍存在于动物组织中，具有重要的生理功能。近年来，由于分子生物学等技术的应用，对许多组织的缝隙连接蛋白的特性及其功能的研究取得了很大进展。本文重点介绍脑内缝隙连接蛋白的类型，分布及其生物物理学和药理学特性，并阐述胶质细胞之间、神经元之间缝隙连接的作用，以及缝隙连接与脑发育和癫痫等神经系统疾病的关系。     

Involvement of programmed cell death in preimplantation embryo demise

Fragmentation is frequently observed in animal and human embryosobtained via in-vitro fertilization (IVF), and is known to beassociated with decreased pregnancy rates and poor survivalfollowing cryopreservation. We postulate that embryo fragmentationis a consequence of activated programmed cell death (PCD) andsubsequent apoptosis and discuss evidence of morphological,histological and biochemical features compatible with the occurrenceof PCD in preimplantation embryos. If PCD is an underlying causeof the high incidence of the fragmentation seen in human pre-embryos,it remains to be determined whether this is reflective of thenatural incidence of lethal chromo somes in the human populationor due to the IVF procedure and culture conditions.     

结肠肿瘤细胞株体外培养过程中的自发性凋亡的观察

研究了Ｖ２３５Ｅ、Ｖ２３５Ｌ、Ｖ４１１三个结肠肿瘤细胞株的细胞凋亡现象。ＤＮＡ电泳分析和ＡＯ／ＥＢ荧光染色形态学检查表明，这三个肿瘤细胞株均存在自发性凋亡。提示肿瘤细胞，包括恶变的癌细胞仍可保留与增殖相对立的凋亡过程。肿瘤细胞存在自发性细胞凋亡的观察，有助于更好地认识肿瘤的发生和发展过程。此外，这些细胞株也为进一步研究细胞凋亡的诱导及其调控提供了有用的细胞模型。     

尼氏小体染色方法的改进及其在神经病理学研究中的应用

组织或细胞的染色在病理学诊断、科学研究和教学工作中,都具有非常重要的意义和使用价值。组织切片染色的质量好坏对于医学诊断,科研和教学至关重要。为了更好的研究神经组织,使医学诊断、科研和教学工作更为方便,本文对Toluidine Blue(甲苯胺蓝)染色方法做了一些改进。在传统的甲苯胺蓝染色过程中,仅考虑对细胞核和尼氏小体进行染色,未考虑细胞浆和其他细胞器:而改进后的甲苯胺蓝染色方法在甲苯股蓝染色后用伊红再染色,既考虑对细胞核和尼氏小体进行染色,也对细胞浆进行了染色。结果显示传统的Toluidine Blue染色结果光镜下观察,细胞核和尼氏小体都可见,即尼氏小体为深蓝色,细胞核为蓝色,染色背景为淡蓝色;改进后的染色结果光镜下观察,尼氏小体为紫蓝色,细胞核为蓝色,染色背景为粉红色。可见,改进后的染色方法染出的组织切片比传统的要清晰、美观。随着科学技术的飞速发展,病理学的研究也随之发展,病理技术势必进一步提高,来适应科技的进步和医学的发展。改进的尼氏小体染色法能够使脑组织切片更清楚观察,更有利于医学工作者对神经组织及尼氏小体的研究。