胆绿素改善大鼠脑缺血再灌注损伤的作用机制

罗靖; 高娴玲; 邵建林; 张超; 张琦

doi:10.12259/j.issn.2095-610X.S20201107

胆绿素改善大鼠脑缺血再灌注损伤的作用机制

doi: 10.12259/j.issn.2095-610X.S20201107

罗靖^{1, 2},
高娴玲³,
邵建林¹,
张超⁴,
张琦^1, ,

1.
昆明医科大学第一附属医院麻醉科
2.
云南省第一人民医院麻醉科，云南昆明　650032
3.
普洱市人民医院麻醉科云南普洱　665000
4.
昆明医科大学第一附属医院神经外科，云南昆明　650032

基金项目: 国家自然科学基金资助项目（81760248；81960250）；云南省应用基础研究重点资助项目（2018FA042）；云南省教育厅科学研究基金资助项目（2019J1250）；昆明医科大学研究生创新基金资助项目（2019S030）

详细信息

作者简介:
罗靖（1993～），男，拉祜族，云南普洱人，硕士研究生，住院医师，主要从事免疫炎症与脑缺血再灌注损伤研究工作

通讯作者:
张琦，E-mail：cuteqq@163.com

中图分类号: R743.3
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出版历程
- 收稿日期: 2020-08-12
- 网络出版日期: 2020-12-03
- 刊出日期: 2020-11-25

Mechanism of Biliverdin in Improving Cerebral Ischemia-reperfusion Injury in Rats

1.
Dept. of Anesthesiology, The 1st Affiliated Hospital of Kunming Medical University
2.
Dept. of Anesthesiology，First People’s Hospital of Yunnan Province，Kunming Yunnan 650032
3.
Dept. of Anesthesiology, Pu’er People’s Hospital, Pu’er Yunnan 665000
4.
Dept. of Neurosurgery, The 1st Affiliated Hospital of Kunming Medical University, Kunming Yunnan 650032, China

摘要

摘要: 目的探讨胆绿素对大鼠脑缺血再灌注损伤的保护作用以及机制。方法采用线栓法建立大鼠大脑中动脉栓塞模型（middle cerebral artery occlusion model，MCAO），给予胆绿素治疗后，采用改良版神经功能缺损评分评价大鼠神经功能缺损情况，干湿重法测定脑组织含水量，TTC染色测定脑梗死面积，Iba-1/DAPI染色标记缺血侧脑组织小胶质细胞活化情况，免疫荧光染色检测脑组织炎症因子TNF-α表达，Western blot测定脑组织Iba-1/CD68的蛋白表达。结果和IR组比较，胆绿素治疗可改善MCAO大鼠的神经功能缺损（P < 0.05），同时，还能减轻脑组织含水量（P < 0.05），降低大脑梗死面积（P < 0.05），以及降低缺血侧脑组织中小胶质细胞的表达和Iba-1/CD68蛋白的表达（P < 0.05）。此外，胆绿素治疗还能减少缺血侧脑组织中炎症因子TNF-α的表达（P < 0.05）。结论胆绿素在MCAO大鼠中可能通过抑制小胶质细胞活化而减轻大鼠脑缺血再灌注损伤。
- 胆绿素 /
- 小胶质细胞 /
- 脑缺血再灌注损伤
Abstract: Objective To investigate the protective effect and underlying mechanism of biliverdin on cerebral ischemia-reperfusion injury in rats. Methods The middle cerebral artery occlusion model（MCAO）was established by inserting nylon thread, after biliverdin treatment, the neurological deficit were evaluated by modified neurological severity score, the brain water content were detected by dry and wet weight method, the cerebral infarction were detected by TTC staining, the activation of microglia in ischemic brain tissue were marked by Iba-1/DAPI staining, the expression of TNF-α were detected by immunofluorescence staining, the protein expression of Iba-1/CD68 were detected by Western blot. Results Compared with the IR group, after biliverdin treatment, the neurological deficits were improved（P < 0.05）, Meanwhile, the brain water content was reduced（P < 0.05）, and the cerebral infarction were attenuated（P < 0.05）. What's more, the microglia expression and Iba-1/CD68 protein expression were decreased（P < 0.05）and the inflammatory factor TNF-α expression was also reduced（P < 0.05）. Conclusion Biliverdin may attenuate cerebral ischemia-reperfusion injury by inhibiting microglia activation in rats.
- Biliverdin /
- Microglia /
- Cerebral ischemia reperfusion injury

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图 1 mNSS评分结果（n = 8）

与Sham组比较，^***P < 0.001；与IR组比较，^#P < 0.05。

Figure 1. The result of mNSS score(n = 8)

下载: 全尺寸图片幻灯片

图 2 大鼠脑组织含水量测定结果

与Sham组比较，^*P < 0.05；与IR组比较，^#P < 0.05。

Figure 2. The results of water content detection in brain tissues of in MCAO rats

下载: 全尺寸图片幻灯片

图 3 胆绿素对MCAO大鼠缺血侧脑梗死面积的影响

A：不同组大鼠脑组织TTC染色结果。B：三组大鼠脑梗死面积比的统计结果。与Sham组比较，^***P < 0.001；与IR组比较，^#P < 0.05。

Figure 3. Effects of Biliverdin on brain infarction volume in MCAO rats

下载: 全尺寸图片幻灯片

图 4 胆绿素对MCAO大鼠缺血侧小胶质细胞活化的影响

A：小胶质细胞在三个实验组中的免疫荧光双染结果。B：Iba-1和CD68在三个实验组中的WB结果；C：Iba-1和CD68在三个实验组中的WB结果分析。与Sham组比较，^**P < 0.01；与IR组比较，^#P < 0.05。

Figure 4. Effects of Biliverdin on activation of microglia in MCAO rats

下载: 全尺寸图片幻灯片

图 5 胆绿素对MCAO大鼠缺血侧小胶质细胞释放TNF-α的影响

A：Iba-1/TNF-α的免疫荧光双染结果；B：TNF-α阳性细胞数比统计结果。与Sham组比较，^***P < 0.001；与IR组比较，^#P < 0.05。

Figure 5. Effect of biliverdin on the release of TNF-α from microglia in MCAO rats

下载: 全尺寸图片幻灯片

表 1 动物分组和处理

Table 1. Animal grouping and treatment

组别	处理	检测指标	数量
Sham组	只暴露颈内动脉，不做其他处理	mNSS/干湿重	n = 8
		TTC染色	n = 4
		WB	n = 4
		免疫荧光双染	n = 8
IR组	缺血2 h再灌注24 h	mNSS/干湿重	n = 8
		TTC染色	n = 4
		WB	n = 4
		免疫荧光双染	n = 8
IR + BV组	缺血2 h再灌注24 h + 胆绿素治疗	mNSS/干湿重	n = 8
		TTC染色	n = 4
		WB	n = 4
		免疫荧光双染	n = 8

下载: 导出CSV

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