一氧化氮诱导大鼠肺动脉平滑肌细胞凋亡机制研究

目的:研究一氧化氮(NO)诱导大鼠肺动脉平滑肌细胞(PASMC)凋亡的作用机制。方法:体外培养Wistar大鼠PASMC,加入NO供体硝普钠(SNP)于常氧和低氧条件下孵育12h或24h,通过流式细胞术碘化丙啶染色法观察PASMC细胞周期时段及凋亡亚二倍体峰变化,应用细胞免疫化学法检测PASMCcaspase-3和NF-κB的表达,同时行DNA断裂琼脂糖凝胶电泳。结果:SNP作用后PASMC出现剂量-依赖性促凋亡作用(P＜0.01),表现为凋亡指数与caspase-3表达的不同程度的增强。随凋亡的进展,出现PASMC凋亡核小体DNAladder,同时PASMCNF-κB阳性核表达较对照组少(P＜0.01)。结论:外源性NO诱导大鼠PASMC凋亡,caspase-3与NF-κB可能涉及PASMC凋亡的调控信号途径。     

CDK4-pRB-E2F1通路是Aβ1-40诱导皮层神经元凋亡的可能途径

目的:探讨β淀粉样肽_1-40(beta-amyloidpeptide_1-40, Aβ_1-40)诱导大鼠皮层神经元凋亡的可能分子机制。方法:以40mg/L的Aβ_1-40诱导离体培养的大鼠皮层神经元凋亡, 流式细胞仪及免疫印迹方法检测细胞周期相关的周期依赖性激酶4(CDK4)、磷酸化的视网膜神经胶质瘤蛋白(pRB)水平;RT-PCR技术检测E2F1mRNA表达水平;荧光分光光度计检测半胱氨酸基天冬氨酸特异性蛋白酶3(caspase-3)活力;观察在Aβ_1-40诱导凋亡过程中是否伴随上述指标的变化。结果:①CDK4、磷酸化pRB水平在Aβ_1-40作用后2-4h显著升高;②Aβ_1-40孵育3h后皮层神经元E2F1基因表达上调;③Aβ_1-40作用12-24h后caspase-3活力明显升高。结论:CDK4-pRB-E2F1信号转导通路可能在Aβ_1-40诱导的大鼠皮层神经元凋亡中起主要作用。     

9602对脑缺血再灌大鼠海马CA1区诱发电位与形态学改变的影响

目的:探讨脑缺血再灌注后海马CA1区诱发群峰电位的变化与形态学改变的关系及中药9602的影响。方法:在整体脑缺血再灌注后不同时间制备的海马脑片上,记录CA1区诱发群峰电位(PS)的变化。采用TUNEL,Nissl染色法进行形态学检测。结果:脑缺血再灌组诱发PS的阈强度明显大于假手术组,波幅显著减小;加条件刺激后PS增幅显著低于假手术组,持续时间缩短。上述变化始于脑缺血再灌后8h,随再灌时间的延长而加重。海马CA1区再灌后8h起TUNEL阳性细胞明显增多,24h达高峰,异常细胞8h最多,随后降低并在低水平持续到7d,细胞总数随再灌时间的延长逐渐减少。中药9602明显降低缺血再灌脑片PS阈强度,增大PS波幅;加大条件刺激后PS增幅并延长持续时间;明显减少海马CA1区的TUNEL阳性细胞数,阻止CA1区细胞数的减少。结论:脑缺血再灌后海马CA1区神经细胞兴奋性和反应性降低,与脑缺血再灌后迟发性神经元死亡(DND)有关,细胞凋亡起重要作用。9602明显改善CA1区神经细胞的兴奋性和反应性,可能与其抑制脑缺血再灌诱导的细胞凋亡,减轻DND的发生有关。     

大鼠松果体衰老的形态学研究

目的 探讨松果体形态结构的衰老性变化,为进一步研究松果体与衰老的关系提供形态学基础。方法 随机选取3月龄和34月龄SD大鼠10只分别作为青年组和老年组,用光镜和电镜结合形态计量学分法分析松果体细胞和神经胶质细胞密度及其指数;松果体细胞线粒体的Vv、Sv、Nv,高尔基器的Vv、Sv、粗面内质网的Sv、溶酶体的Vv;松果体神经终末的Vv及其线粒体的Vv、Sv、Nv、小颗粒囊泡的NA。结果 与青年组相比,老年组松果体细胞核形状不规则,核膜皱褶增多和加深。细胞浆内粗面内质网减少,排列紊乱,分散,脱颗粒明显;线粒体结构不清、肿胀、嵴断裂、消失、空泡化等。老年组松果体细胞密度比青年组减少,而神经胶质细胞密度增加,差异有高度显著性(P<0.01)。老年组神经胶质细胞指数比青年组高(P<0.01)。老年线粒体Vv与青年组无差别,但老年组线粒体Sv、Nv均比青年组减少(P<0.01)。老年组高尔基器的Vv、Sv均比青年组减少(P<0.01);老年组粗面内质网Sv比青年组减少,而老年组溶酶体Vv比青年组增加(P<0.01);老年组神经终末Vv比青年组减少(P<0.01);老年组神经终末中小颗粒囊泡NA比青年组减少(P<0.01);老年组线粒体Sv、Nv比青年组减少(P<0.05),两组神经终末中线粒体Vv无差别(P>0.05)。结论 老年大鼠松果体已发生衰老性变化,这可能与机?     

体外培养诱导大鼠骨髓间质干细胞分化为肌样细胞的实验研究

目的:体外培养并诱导大鼠骨髓间质干细胞(MSCs)分化为肌样细胞。方法:采用常规技术对SD鼠MSCs进行体外培养传代、鉴定、诱导分化和免疫组化、透射电镜检测分析。结果:流式细胞仪检测, 细胞表达CD29和CD44, 不表达CD11b和CD45;经一定浓度5-氮杂胞苷和两性霉素B诱导分化后细胞desmin和myoglobin染色阳性;电镜观察肌样细胞胞浆靠胞膜缘可见无细胞器的条状肌丝区带。结论:传代贴壁生长的梭形细胞为MSCs。MSCs可能具有表达肌细胞的特异性启动或分化调控基因;5-氮胞苷等化合物可使DNA的胞嘧啶去甲基化, 从而诱导MSCs向肌源性细胞分化。临床有运用MSCs治疗肌萎缩性疾病的前景。     

Effect of treadmill exercise on food intake and body weight in lean and obese rats

Body weight and food intake of lean and obese, male and female Osborne-Mendel rats following treadmill exercise were compared. Rats were assigned, separately by sex, to one of three diet groups; Group 1 was fed a low fat (10%) diet throughout the study, Group 2 was fed a high fat (55%) diet for 16 weeks and then switched to the low fat diet 1 week prior to exercise, and Group 3 was fed the high fat diet throughout the study. To control for differences in work output between the leanest and heaviest animals, exercise intensity was adjusted across groups such that all exercised rats had equivalent energy expenditure. After a 3 day training period, the exercise was successively increased over 8 days until a work output of 374.9J was reached. Relative to their respective controls, obese exercised males showed a reduction in body weight but no change in food intake. In contrast, exercised females showed no change in body weight or food intake, regardless of dietary condition.     

溶酶体在实验性心肌肥大过程中的作用

本实验用大鼠主动脉结扎的心肌肥大模型,结合电镜酶细胞化学技术,观察心肌肥大发展过程中溶酶体的变化。结果表明:心肌肥大发展时,溶酶体参与心肌细胞改建;到心肌肥大后期,出现大量自噬现象,造成心肌不可逆损伤。     

肾炎益气液对肾毒血清性肾炎大鼠肾脏超微结构的影响

目的:观察肾毒血清性肾炎大鼠肾组织超微结构变化及肾炎益气液对该变化的影响。方法: 采用大鼠肾毒血清性肾炎模型,常规电镜制片、染色,观察肾组织超微结构的变化。结果: 注射肾毒血清(NTS)后,病理组大鼠电镜下可见明显的系膜细胞增生,系膜基质增多,毛细血管腔闭塞及上皮下、内皮下电子致密物沉积等多种病理损伤性变化。而肾炎益气液组上述病变有所减轻。结论: 肾炎益气液有不同程度减轻肾小球电镜下病变的作用。     

The effects of age and strain on aggression in male rats

Aggressive behaviors were compared for male Fischer-344 and Long-Evans rats formed into mixed-sex colonies at 100, 200, or 300 days of age. Within each strain, male aggression did not differ across these ages, nor did it differ for an additional group of 600-day old Long-Evans males: both the form and the incidence of specific aggressive and defensive behaviors remained relatively constant over an age range representing nearly the entire normal life span for this species. However, Fischer-344 males at all ages were reliably lower in aggression than the Long-Evans rats. The specific aggression components seen in the Fischer-344 rats tended to be nondamaging, resembling “play fighting” more than the tissue damaging attack behaviors typical of other strains. Despite the frequent use of this strain in gerontological research, such low levels of aggressiveness counterindicate their use in research in aging and aggression.     

Further studies on the aberrant crossed visual corticotectal pathway in rats

Summary An aberrant crossed corticotectal pathway can be generated by removal of one visual cortex and the contralateral superior colliculus from newborn rats. This aberrant crossed corticotectal projection arises from the pyramidal neurons located in layer V of the visual cortex and terminates in a spatially orderly manner in the appropriate laminae of the cortically deafferented contralateral colliculus. Comparable results cannot be reproduced by unilateral collicular lesions alone. The significance of these findings and the possible mechanisms involved in the formation of the aberrant pathway are discussed and compared with the retinotectal system.The research was supported by USPHS Grant EY-00596 from the National Institutes of Health