杂色曲霉素对小鼠室周器官及其它脑区的影响及其机制的研究

发布时间：2018-01-23 14:27

  本文关键词： 杂色曲霉素 室周器官 超微结构 TNF-α TGF-β2 凋亡　出处：《河北医科大学》2006年博士论文　论文类型：学位论文

【摘要】： 目的:霉菌毒素对脑组织的损伤受到了国内外许多学者的广泛关注,但是,杂色曲霉素(Sterigmatocystin, ST)对脑组织的影响国内外尚未见报道。室周器官(circumventricular organs, CVOs)是脑室周围的一些微小器官,包括穹隆下器、正中隆起、终板血管器、脉络丛、连合下器、松果体后叶及最后区等,由于其缺乏血脑屏障,因此是监控血成分的理想位点。本实验通过灌胃给药构建ST动物实验模型,研究给药后不同时间点ST对CVOs生物学效应。观察给药后小鼠穹隆下器、脉络丛和外侧隐窝形态学变化及其室管膜和室管膜下细胞超微结构变化。检测小鼠穹隆下器、脉络丛和外侧隐窝TNF-α、TGF-β2基因的表达情况。检测小鼠穹隆下器、脉络丛和外侧隐窝和其他脑区细胞凋亡,探讨ST对脑组织的影响及其机制。其意义在于首次研究ST毒素对CVOs的作用,为以后更加深入了解ST毒素对脑组织的影响奠定了一定的基础,同时为研究其他毒素对脑组织的影响开辟了一条新的途径。 方法: 1扫描电镜和透射电镜检查 雄性BALB/c小鼠,体重18~20 g,由河北医科大学实验动物部提供。将18只BALB/c雄性小鼠随机分为6组,每组3只。处理组5组,对照组1组,处理组按ST 3000μg/kg(sigma公司)的剂量,溶于0.5 ml生理盐水中灌胃。对照组1组用等量的生理盐水灌胃。灌胃后即刻处死动物,处理组动物于灌胃后1、2、4、8、16 h处死动物。 1.1扫描电镜标本制备 麻醉动物后,开胸经心脏常规灌注,灌注液为1.5%多聚甲醛和2.5%戊二醛磷酸缓冲液(0.1 mol/L,pH7.2)。灌注完毕取出整脑,在固定剂中(4℃)后固定12 h。将修好的脑组织块用磷酸缓冲液(0.1 mol/L,pH7.2)仔细冲洗3次,再放入磷酸缓冲液中过夜,梯度酒精脱水,叔丁醇置换,二氧化碳临界点干燥,真空喷金,日立S-3500N扫描电镜观察、照像。
[Abstract]:Objective: the damage of mycotoxins to brain tissue has been widely concerned by many scholars at home and abroad, but Sterigmatocystin. The effects of ST) on brain tissue have not been reported at home and abroad. Periventricular organ circumventricular organs (CVOss) are some small organs around the ventricle. These include the subfornix, the median eminence, the endplate vascular apparatus, the choroid plexus, the subconjunctival apparatus, the posterior lobe of the pineal gland and the final area, due to its lack of blood-brain barrier. Therefore, it is an ideal site to monitor the blood composition. In this experiment, St animal model was established by intragastric administration, and the biological effects of St on CVOs were studied at different time points after administration, and the subfornial organ of mice was observed after administration. Morphological changes of choroid plexus and lateral recess and ultrastructural changes of ependymal and subependymal cells were observed. Expression of TGF- 尾 2 gene. Apoptosis of mouse subfornical organ, choroid plexus, lateral recess and other brain regions were detected. The purpose of this study is to study the effect of St toxin on CVOs for the first time, and to lay a foundation for further understanding the effect of St toxin on brain tissue. At the same time, it provides a new way to study the effects of other toxins on brain tissue. Methods: 1 scanning electron microscopy and transmission electron microscopy Male BALB/c mice, weighing 1820 g, were provided by the experimental animal department of Hebei Medical University. Eighteen BALB/c male mice were randomly divided into 6 groups, 3 in each group and 5 in treatment group. In the control group 1, the dose of ST3000 渭 g / kg sigma was used in the treatment group. The rats in the control group were given the same amount of normal saline. The animals in the treatment group were killed immediately after gastric perfusion, and the animals in the treatment group were killed at 816 hours after 1 hour of gastric perfusion. 1.1 preparation of SEM specimens After anesthesia, the chest was routinely perfused with 1.5% paraformaldehyde and 2.5% glutaraldehyde phosphate buffer (0.1 mol / L 路L ~ (2 +)). The whole brain was taken out after perfusion. After fixation at 4 鈩，

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