早期活体判断新生小鼠缺氧-缺血性脑损伤程度

颜琳; 张如秋; 华海蓉; 王茜; 曾祥菲; 李娟娟; 江萍; 李凡

doi:10.16098/j.issn.0529-1356.2021.05.001

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解剖学报 ›› 2021, Vol. 52 ›› Issue (5) : 675-680. DOI: 10.16098/j.issn.0529-1356.2021.05.001

神经生物学

早期活体判断新生小鼠缺氧-缺血性脑损伤程度

颜琳¹ 张如秋¹ 华海蓉¹ 王茜¹ 曾祥菲¹ 李娟娟² 江萍^1* 李凡^1,3*

作者信息 +

Judging the degree of brain injury after hypoxia-ischemia in living neonatal mice in the early stage

YAN Lin¹ ZHANG Ru-qiu¹ HUA Hai-rong¹ WANG Qian¹ ZENG Xiang-fei¹ LI Juan-juan² JIANG Ping^1* LI Fan^{1, 3*}

Author information +

文章历史 +

摘要

目的探讨早期活体区分缺氧-缺血性脑损伤（HIBD）程度的方法，为后续研究不同程度HIBD的分子机制奠定基础。方法采用改良Rice-Vannucci法构建C57BL/6 J小鼠HIBD模型。模型建立后1 d和3 d经活体剪开小鼠头皮行脑组织大体观察，区分轻重损伤。其后运用2, 3, 5-氯化三苯基四氮唑（TTC）染色、激光散斑脑血流成像、HE染色、Fluoro-Jade B（FJB）染色及术前、术后体重差，验证活体观察脑损伤程度的可靠性。结果 HIBD后1 d和3 d经活体剪开小鼠头皮行脑组织大体观察，可将其区分为轻度损伤（HI-M）组和重度损伤（HI-S）组。HI-S组损伤侧出现明显的梗死灶，脑血流量显著减少，大量神经元坏死，体重较术前显著降低。HI-M组损伤侧脑血流量仅在HIBD后3 d减少，形态学观察到损伤侧皮层和海马区神经元排列疏松，体重较术前降低。结论剪开头皮行脑组织大体观察是早期活体情况下区分HIBD轻度及重度脑损伤的可靠方法。

Abstract

Objective To explore the method of distinguishing the degree of hypoxic-ischemic brain damage (HIBD) in living mice in early stage, so as to lay a foundation for the follow-up study of the molecular mechanism of different degrees of HIBD. Methods The modified Rice-Vannucci method was used to duplicate the HIBD model of C57BL/6 J mice. On the 1 day and 3 days after the model, the scalp of mice were cut and the brain tissue were observed to distinguish between mild and severe lesions in living mice, and then 2, 3, 5-triphenyltetrazolium chloride (TTC) staining, laser speckle cerebral blood flow imaging, HE staining, Fluoro-Jade B (FJB) staining and body weight difference before and after operation were used to verify the reliability of observation in living mice.

Results Through the gross observation of brain tissue in living mice, HIBD could be divided into mild injury (HI-M) group and severe injury (HI-S) group. On day 1 and day 3 after HIBD, a significant decrease in cerebral blood flow, obvious gray infarction and a large number of necrotic neurons were observed in the HI-S group, and the body weight was significantly lower than that before operation. In the HI-M group, the cerebral blood flow of the injured side decreased only on the 3rd day after HIBD, and the loose arrangement of neurons in the cortex and hippocampus of the injured side was observed morphologically. The body weight was lower than that before operation. Conclusion Gross observation of brain tissue by cutting the scalp is a reliable method to distinguish mild and severe brain injury in the early stage of HIBD in living mice.

导出引用

颜琳张如秋华海蓉王茜曾祥菲李娟娟江萍李凡. 早期活体判断新生小鼠缺氧-缺血性脑损伤程度[J]. 解剖学报. 2021, 52(5): 675-680 https://doi.org/10.16098/j.issn.0529-1356.2021.05.001

YAN Lin ZHANG Ru-qiu HUA Hai-rong WANG Qian ZENG Xiang-fei LI Juan-juan JIANG Ping LI Fan. Judging the degree of brain injury after hypoxia-ischemia in living neonatal mice in the early stage[J]. Acta Anatomica Sinica. 2021, 52(5): 675-680 https://doi.org/10.16098/j.issn.0529-1356.2021.05.001

中图分类号： R36

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