论文部分内容阅读
目的建立一种新的鼠脑局部蛛网膜下腔注射氧合血红蛋白(OxyHb)的动物模型,探讨OxyHb在体诱导鼠脑细胞凋亡空间分布及时间依赖性规律的研究。方法采集ICR鼠新鲜血液制备OxyHb,建立小鼠脑局部蛛网膜下腔注射OxyHb动物模型。动物被分为实验组(40只)及对照组(35只),实验组注射OxyHb(50μL),对照组注射生理盐水(50μL)于鼠脑蛛网膜下腔,再按处死时间将小鼠分为3h、6h、12h、24h和48h亚组。采用HE染色、透射电子显微镜及原位细胞凋亡检测法(TUNEL)三种技术辨别细胞坏死或凋亡。结果实验组3h OxyHb集中分布于注射点同侧脑表面,6～12h OxyHb积聚于脑基底池及交叉池．24～48h OxyHb已被吸收；实验组鼠脑细胞凋亡主要分布于OxyHb注射区同侧脑皮质及双侧海马区；经HE染色、透射电子显微镜观察显示鼠脑细胞发生凋亡形态学改变；实验组TUNEL阳性细胞较对照组明显增多,且随时间延长凋亡细胞逐渐减少。结论本实验建立的鼠脑局部蛛网膜下腔注射OxyHb的动物模型,方法较简单,重复性好。OxyHb在体可诱导鼠脑细胞凋亡而非坏死,凋亡细胞主要分布于OxyHb注射区同侧脑皮质及双侧海马区,且随时间延长而凋亡细胞逐渐减少,具有空间分布和时间依赖性的规律。
Objective To establish a new animal model of subarachnoid injection of oxygenated hemoglobin (OxyHb) in mice and to investigate the spatial distribution and time-dependent regulation of apoptosis of OxyHb in vivo. Methods Fresh blood of ICR mice was collected to prepare OxyHb, and the animal model of OxyHb injection into the subarachnoid space was established. The animals were divided into experimental group (40) and control group (35). OxyHb (50μL) was injected into the experimental group and normal saline (50μL) to the subarachnoid space of the mouse brain. 3h, 6h, 12h, 24h and 48h subgroups. HE staining, transmission electron microscopy and in situ cell apoptosis assay (TUNEL) were used to identify cell necrosis or apoptosis. Results OxyHb in the experimental group was concentrated on the ipsilateral brain surface at 3h after injection, and accumulated in brain basal cistern and cross pool in 6 ~ 12h. 24 ~ 48h OxyHb has been absorbed; experimental group of rat brain apoptosis mainly in the ixyHb injection area ipsilateral cerebral cortex and bilateral hippocampus; by HE staining, transmission electron microscopy showed morphological changes of apoptotic brain cells; TUNEL-positive cells in the experimental group were significantly increased compared with the control group, and apoptotic cells gradually decreased with time. Conclusion The animal model of OxyHb injected into the subarachnoid space of mouse brain established in this experiment is simple and reproducible. OxyHb can induce apoptosis in rat brain cells rather than necrosis. Apoptotic cells are mainly distributed in ipsilateral cerebral cortex and bilateral hippocampus of OxyHb injection area, and the apoptotic cells gradually decrease with time, with spatial distribution and time-dependent Sexual patterns.