循环间歇性低氧对大鼠肾上腺β淀粉样前体蛋白裂解酶1表达的影响

目的观察β淀粉样前体蛋白裂解酶1(BACE1)在大鼠肾上腺的表达和定位,检测循环间歇性低氧(CIH)对BACE1表达的影响。方法采用Western blotting和免疫组织化学法检测BACE1在大鼠肾上腺的表达及定位;16只雄性SD大鼠随机分为2组,常氧对照组(control组)和CIH模型组,每组8只。模型制作2周,Western blotting检测各组大鼠肾上腺髓质BACE1和酪氨酸羟化酶(TH)蛋白的表达量。结果BACE1主要定位于大鼠肾上腺髓质神经纤维;与control组相比,CIH组大鼠肾上腺髓质BACE1蛋白水平下降,TH蛋白水平升高(P<0.05)。结论BACE1定位于大鼠肾上腺髓质神经纤维;BACE1水平下调可能参与减缓CIH引起的交感神经活性过度增强。     

糖皮质激素对大鼠下丘脑亨廷顿蛋白相关蛋白1表达的影响

目的 探讨外源性糖皮质激素对大鼠下丘脑亨廷顿蛋白相关蛋白1（HAP1）表达的影响。 方法 成年雄性Wistar大鼠80只,随机分为实验对照组（注射生理盐水）、皮质酮注射组、皮质酮+皮质酮受体拮抗剂(RU38486)注射组、饮用氢化可的松组。皮质酮注射［(5mg/(kg.12h)］组又分为注射持续1d、3d、5d、7d组;皮质酮+皮质酮受体拮抗剂(RU38486)注射［(10mg/(kg.12h)］组注射持续3d;饮用氢化可的松组大鼠饮用水中加氢化可的松（0.01g/L）,连续1个月。应用RT-PCR、免疫印迹法观察大鼠下丘脑HAP1表达的变化。 结果 注射皮质酮1d和3d后,下丘脑HAP1表达明显减少,其mRNA的表达明显下调,5d后HAP1开始回升,7d 后恢复到正常水平,HAP1 mRNA表达5d后明显增加,并高于正常水平;长期(一个月)饮用氢化可的松的大鼠,其下丘脑中HAP1的表达明显减少;在皮质酮注射的同时注射RU38486,可以对抗由皮质酮注射所引起的下丘脑HAP1的变化。 结论 外源性过量糖皮质激素能影响下丘脑HAP1的表达,下丘脑中的HAP1可能参与下丘脑神经元或神经内分泌细胞的功能活动。     

链脲佐菌素敏感的胰岛细胞表达亨廷顿蛋白相关蛋白1

目的探讨亨廷顿蛋白相关蛋白1（HAP1）在大鼠胰岛中的定位.方法应用一次性大剂量腹腔注射链脲佐菌素（streptozotocin）的方法选择性破坏大鼠胰岛B细胞复制糖尿病模型,免疫组织化学ABC法显示胰岛内HAP1和胰岛素免疫反应性.结果在正常大鼠胰腺内,HAP1选择性表达于胰岛内,HAP1免疫反应阳性细胞呈短条索状或团状分布在每个胰岛内,主要位于胰岛的中央部,与胰岛B细胞的分布十分相似.注射链脲佐菌素的大鼠,胰岛中含HAP1的细胞数量在注射链脲佐菌素3d后已明显减少,并随着注射后动物存活时间的延长而进一步进行性减少;在注射后4周的大鼠,胰岛中仅有少数散在分布的HAP1免疫反应弱阳性细胞.链脲佐菌素对胰岛中表达HAP1细胞的影响与对表达胰岛素细胞的影响一致.结论HAP1也存在于胰岛内,并主要定位于链脲佐菌素敏感的B细胞内.     

慢性复合应激对大鼠海马生长休止蛋白7表达的影响

目的 探讨慢性复合应激对大鼠海马神经元中生长休止蛋白7(Gas7)的表达变化及意义.方法 36只大鼠随机分为慢性复合应激组和正常对照组.复合应激组动物进行6周的垂直旋转、睡眠剥夺、捆绑(6h/d)和夜间光照等慢性复合性应激试验;实验结束后,所有动物采用免疫组织化学和免疫印迹等方法 检测大鼠海马Gas7蛋白表达的变化和神经元凋亡情况.结果 Gas7在对照组和实验组大鼠海马各区均有表达,主要表达在海马神经元的胞质和突起内.其中在CA1区和齿状回呈较强阳性表达,CA3区表达则较弱;慢性复合应激组CA1区和齿状回阳性染色加深,平均吸光度明显上升(P<0.05),而CA3区则不明显(P>0.05).慢性复合应激组中部分切片在下托可见少量Caspase-3免疫反应阳性细胞.免疫印迹检测表明,慢性复合应激组大鼠海马Gas7表达明显增强,与正常对照组相比,差异具有显著性(P<0.05).结论 Gas7可能参与了在应激情况对神经元的保护作用,以及促进神经元的发生和发育.     

自发性高血压大鼠交感神经节与肾上腺髓质内NPY的表达及超微结构的变化

目的:通过对自发性高血压大鼠(SHR)颈上神经节、肾上腺髓质内神经肽酪氨酸(NPY)表达及超微结构变化的研究,探讨NPY在高血压发生和发展中的作用。方法:随机选取成年大鼠60只,分成SHR组和正常对照组,每组各30只,采用Real-Time PCR技术和免疫组织化学法,检测两组大鼠颈上神经节、肾上腺髓质内NPYmRNA和蛋白的表达;应用透射电镜技术观察上述两种组织内超微结构的改变。结果:与同周龄的正常组大鼠相比,SHR大鼠的血压明显增高(P<0.05),RT-PCR和免疫组织化学方法检测到颈上神经节、肾上腺髓质内NPYmRNA和蛋白质的表达均增高(P<0.05),NPY免疫阳性神经元的数量和阳性细胞的光密度亦增加(P<0.05)。电镜结果示SHR大鼠两种组织内异染色质、线粒体、粗面内质网等数量均明显增加(P<0.05)。结论:SHR颈上神经节、肾上腺髓质内的NPY通过复杂的机制可能参与了高血压的形成。     

亨廷顿蛋白相关蛋白1在大鼠脊髓中的定位及与初级传入神经元的关系

本文首次通过免疫组织化学技术和免疫印迹方法(Western blotting)观察了亨廷顿蛋白相关蛋白1(huntingtin associated protein 1，HAP1)在大鼠脊髓和背根节中的分布以及HAP1与Gfiffonia simplicifolia Ⅰ-B4结合位点在背根节初级传入神经元的共存，并观察了单侧背根切断和辣椒素(capsaicin)损毁实验对脊髓背角HAP1表达水平的变化。     

亨廷顿蛋白相关蛋白-1在成年大鼠脑内的定位

目的:探讨亨廷顿蛋白相关蛋白1(HAP1)在成年大鼠脑内的分布.方法:健康成年大鼠取脑做连续冠状切片,免疫组织化学方法观察HAP1在脑内的表达.结果:大量的HAP-1表达出现在海马、齿状回、隔核、黑质、红核、连合下核、连合下器、脊髓前庭核、臂旁核、大脑脚基底部、束间核、后屈束、穹窿、被盖腹侧区、嘴侧线形中缝核以及丘脑、下丘脑各区;蜗背侧核、背侧纵束、前庭内侧核、中缝背核背侧部、中缝背核腹侧部、中缝背核腹外侧部、中央灰质等处的表达稍弱;其余像嗅球、大脑皮层各区、小脑皮层等处均有表达但强度极低,Stigmoid小体数目的变化趋势与HAP1基本一致.结论:HAP1广泛分布于脑的各部,推测HAP1可能参与了脑的多种生理功能.     

海洛因戒断及复吸对大鼠肾上腺髓质细胞β-内啡肽和促肾上腺皮质激素表达及血清皮质醇的影响

目的 探讨海洛因戒断、脱毒及复吸对大鼠肾上腺髓质细胞β-内啡肽（β-EP）和促肾上腺皮质激素（ACTH）表达及血清皮质醇（COR）的影响。方法 成年雄性SD大鼠63只,随机分为实验组（21只）、盐水对照组（21只）和正常对照组（21只）,实验组又分为戒断组、脱毒组和复吸组,采用免疫组织化学、图像分析法及放射免疫测定法进行研究。结果 与正常及盐水对照组比较,海洛因戒断组β-EP免疫反应（β-EP-IR）细胞染色变浅,平均灰度值升高（P <0.05）;经美沙酮脱毒治疗,β-EP-IR细胞染色较正常及盐水对照组加深,平均灰度值降低（ P <0.05）;海洛因复吸组β-EP-IR细胞表达再次减弱,但与正常及盐水组比较,平均灰度值差异无显著性。与正常及盐水对照组比较,实验各组大鼠肾上腺髓质ACTH免疫反应（ACTH-IR）细胞染色均变浅,平均灰度值增高,有显著差异（P <0.05）。放射免疫测定结果表明,实验各组大鼠血清COR均显著高于正常对照组（P <0.05）。结论 海洛因戒断、脱毒、复吸影响了大鼠肾上腺髓质细胞表达β-EP、ACTH以及血清COR的分泌。     

生脉注射液联合去甲肾上腺素对多器官功能障碍综合征大鼠淋巴细胞凋亡及凋亡蛋白的影响

目的:研究生脉注射液联合去甲肾上腺素对多器官功能障碍综合征大鼠细胞免疫的影响。方法:选取40 只健康雄性大鼠,通过应用酵母多糖腹腔注射致MODS 模型,采用数字表法随机分为对照组、生脉注射液组、去甲肾上腺素组以及生脉注射液联合去甲肾上腺组, 每组10 只。用药后6、24、48 h 检测血清中TNF鄄琢、IL鄄1茁、IL鄄12 和IL鄄2 的浓度,免疫组化检测大鼠脾脏caspase鄄3、caspase鄄8、caspase鄄9 活性,流式细胞技术测定淋巴细胞凋亡情况及Th1/ Th2。结果:淤与对照组相比,生脉注射液组、去甲肾上腺组以及联合用药组,TNF、IL鄄1茁浓度上升不是很明显,且随着治疗时间的延长,治疗各组TNF、IL鄄1茁浓度逐渐下降,联合用药组TNF、IL鄄1茁基本趋于正常;而IL鄄12、IL鄄2 趋势则相反,治疗各组较对照组血清IL鄄12、IL鄄2 浓度明显升高,联合用药组升高更加明显。于与对照组相比,生脉注射液组、去甲肾上腺素组以及联合用药组caspase鄄3、caspase鄄8、caspase鄄9 表达均显著减少,且联合用药组凋亡蛋白的表达下降更明显,差异有统计学意义(P<0郾05)。盂生脉注射液组、去甲肾上腺素组以及联合用药组淋巴细胞凋亡的阳性细胞数比率较对照组明显减少,联合用药组淋巴细胞凋亡的阳性细胞数减少较生脉注射液组以及去甲肾上腺素组更明显;相比于对照组,生脉注射液组、去甲肾上腺素组以及联合用药组Th1/ Th2 比率明显增高,联合用药组增高更明显,差异有统计学意义(P<0.05)。结论:生脉注射液联合去甲肾上腺素对多器官功能障碍大鼠具有减轻炎症反应和改善免疫功能的作用。     

睡眠剥夺后大鼠皮质、海马和杏仁核微管相关蛋白-2及神经丝蛋白的表达变化

目的探讨睡眠剥夺后大鼠皮质、海马和杏仁核微管相关蛋白-2(MAP-2)及神经丝蛋白(NF-200)表达的变化。方法采用改良小平台水环境法制作大鼠睡眠剥夺模型,大鼠随机分为睡眠剥夺组(SD组)、环境对照组(TC组)和空白对照组(CC组)。SD组包括睡眠剥夺6h、12h、1d、2d、3d、5d、7d共7个时点,每个时间点5只大鼠,CC组5只。免疫组织化学法观察MAP-2和NF-200阳性表达。结果睡眠剥夺5d和7d时,皮质、CA1、CA2、CA3、齿状回和杏仁核MAP-2和NF-200阳性表达减少(P0.05,P0.01)。结论睡眠剥夺可导致脑组织MAP-2和NF-200表达减少。     

Chromaffin cells in the adrenal homolog of Aphanius fasciatus (teleost fish) express piecemeal degranulation in response to osmotic stress: a hint for a conservative evolutionary process

The effect of severe osmotic stress on the ultrastructural morphology of chromaffin cells in the adrenal homolog of Aphanius fasciatus, a small eurhyaline teleost living in saltpans, was evaluated by electron microscopy quantitative analysis. Fishes were transferred from salt water, whose salinity was 3.7%, to dechlorinated tap water and chromaffin cells were studied at resting condition and after 2 and 48 hr from the beginning of the experiment. Ultrastructural examination revealed a series of granule and cytoplasmic changes highly specific for piecemeal degranulation (PMD), a secretory process based on vesicular transport of cargoes from within granules for extracellular release, which was previously described in chromaffin cells of the mouse, rat, and human adrenal medulla. There was indeed a significant trend toward loss of content material from chromaffin granules accompanied by enlargement of granule size. Remarkably, chromaffin granules maintained their individual close structure during the whole releasing process and eventually transformed into large empty containers. A dramatic increase in the density of small, membrane-bound, variably electron-dense vesicles free in the cytoplasm or attached to granules was recognized during the first 2 hr of stress response. These features fell to control levels after 48 hr. A similar time-course pattern was observed concerning the formation of budding projections from the surface of chromaffin granules. This study provides new insight into PMD physiology and suggests that PMD is part of an adaptive secretory response to severe osmotic stress in fishes. From an evolutionary point of view, this study lends support to the concept that PMD is a secretory mechanism highly conserved throughout vertebrate classes.     

PCNA在高血压大鼠肾上腺的表达

目的　观察Dahl大鼠肾上腺中增殖细胞核抗原 (PCNA)表达的动态变化 ,探讨原发性高血压肾上腺细胞增殖的关系。方法　应用日本引进的Dahl高血压大鼠 ,SP免疫组织化学染色。结果　PCNA免疫阳性细胞在Dah1S和Dah1R大鼠肾上素均有表达 ,主要存在于皮质中。阳性反应产物见于细胞核中。PCNA的阳性细胞数 ,在盐敏感性高血压 (Dah1S)大鼠 ,随着高血压的进展而呈先升高后降低趋势。结论　Dah1高血压可导致大鼠肾上腺皮质PCNA表达的动态变化 ,先升高后降低。初期的增高可能与肾素 血管紧张素 醛固酮系统活性增高有关。后期则可能与持续的高钠离子负荷有关。     

家兔脑膜炎双球菌感染性休克的肾上腺病变

目的:探讨家兔被脑膜炎双球菌(MC)攻击后肾上腺组织的病变特征。方法:采用MC-A群菌珠,按1×10¹¹菌体/kg体重剂量,清醒动物iv(模型组,n=10)复制成动物模型;并以生理盐水为对照(对照组,n=8)。给药前、给药后60min\,120min分别检查血浆TNF-α水平,120min后处死动物,取肾上腺做TNF-α细胞组化和超微结构检查。结果:模型组髓质区内可见TNF-α阳性细胞、TNF-α阳性物质释放入髓质并引起髓质坏死。肾上腺皮质细胞无病变。血浆TNF-α浓度增高。结论:MC攻击家兔后,可引起TNF-α在肾上腺髓质释放并引起病变,而早期肾上腺皮质腺细胞无损伤。     

创伤后应激障碍大鼠海马神经元凋亡中Caspase-12的表达

目的观察创伤后应激障碍(PTSD)大鼠海马神经元凋亡和Caspase-12表达的变化。方法采用单一延长应激(SPS)方法刺激大鼠建立PTSD模型,健康雄性Wistar大鼠随机分为SPS 1d、4d、7d组和正常对照组。应用透射电子显微镜观察PTSD大鼠海马神经元的形态学改变,采用免疫组织化学、免疫印迹法和RT-PCR技术检测海马神经元Caspase-12表达的变化。结果 SPS后,海马神经元发生了凋亡的形态学改变,Caspase-12的表达于刺激后4d最多。结论 PTSD激活凋亡因子Caspase-12启动海马神经元经内质网途径细胞凋亡,可能是PTSD患者海马萎缩的一个病理生理学基础。     

创伤后应激障碍大鼠海马神经元凋亡的形态学变化及相关基因表达

目的 探讨创伤后应激障碍(PTSD)大鼠海马神经元Bcl-2和Bax的表达及与神经元凋亡的关系.方法 采用国际认定的SPS方法刺激大鼠建立PTSD大鼠模型,取SPS刺激后1d、4d、7d、14d组和正常对照组.应用透射电镜和原位末端标记法观察PTSD大鼠海马神经元细胞凋亡的形态学变化并检测细胞凋亡指数;应用免疫组织化学、免疫荧光双标、激光共焦显微镜技术和免疫印迹法检测Bel-2和Bax蛋白的表达. 结果 PTSD大鼠海马神经元出现细胞凋亡改变;凋亡细胞数量随时问的延长而逐渐增多,于SPS刺激后7d达高峰;Bel-2蛋白于SPS刺激后4d达高峰;Bax蛋白于SPS刺激后7d达高峰,之后逐渐下降.Bcl-2/Bax的比值于SPS刺激后一过性增加,以后随时间延长比值下降. 结论海马神经元细胞凋亡可能是PTSD大鼠海马萎缩的原因之一;Bel-2和Bax蛋白含量增加以及两者的比值失衡可能是导致PTSD大鼠海马神经元凋亡的原因之一.     

Structure of the adrenal gland in rat strain with hereditary stress-induced arterial hypertension nursed by Wistar dams

The comparative investigation of the adrenal structure in two groups of rats of Inherited Stress-Induced Arterial Hypertension (ISIAH) strain was conducted. The animals of the first group were nursed by normotensive Wistar rats, while those of the second (control) group were reared by their own mothers. The volume of the adrenal medulla in rats of the first group was found to exceed that in the second group. In the adrenal zona glomerulosa and zona fasciculata of 3-week-old rats of the first group parenchymal-stromal ratio and the average volume of adrenocorticocytes were lower than those in the animals of the second group. This was accompanied by a decrease in mitochondrial volume density and accumulation of lipid droplets in the cytoplasm, indicating a reduction of hormone-producing activity of endocrinocytes in the animals the first group as compared to controls. By 6 month of age, arterial pressure and quantitative parameters of adrenal medulla and cortex in the animals the first group were..... as compared to those in the second group. It is suggested that the nursing of ISIAH rat pups by normotensive Wistar dams had modulating effect on the adrenal structure and therefore on arterial hypertension development.     

Morphological correlates of hormone secretion in the rat adrenal cortex and the role of filopodia

Rat adrenals in different states of stimulation were examined by transmission electron microscopy following perfusion fixation using an in situ isolated-circulation technique. In unstimulated glands, intracortical capillaries were constricted and the cells of the cortex were pressed closely together with little development of filopodia or intercellular spaces. Glands fixed during the period of operative stress, or following a 1 hr perfusion with Adrenocorticotropic hormone (ACTH) showed that the radially orientated capillaries of the cortex were massively expanded, and the cells of both the glomerulosa and fasciculata exhibited an extensive development of filopodia on their surfaces. These filopodia extended into large intercellular spaces, where they often entered into complex relationships with filopodia from neighboring cells. The development of filopodia by cells of the adrenal cortex was also observed using scanning electron microscope techniques. In cells either incubated with ACTH in vitro or isolated from adrenals of rats treated with ACTH in vivo, the filopodia were numerous, often branched, and could reach as much as 1 micro m in length. In contrast, adrenal cells obtained from animals pretreated with cortisol were smooth surfaced. Other cell characteristics, including mitochondria, smooth endoplasmic reticulum, dense granules, and coated vesicles did not show such dramatic correlations with the state of stimulation. It is considered that the development of filopodia and intercellular space is related to secretory mechanisms in the rat adrenal cortex.