稳定内皮和周细胞微管改善脊髓损伤中微循环障碍的作用及其机制

doi:10.16098/j.issn.0529-1356.2021.01.003

解剖学报 ›› 2021, Vol. 52 ›› Issue (1): 21-29.doi: 10.16098/j.issn.0529-1356.2021.01.003

稳定内皮和周细胞微管改善脊髓损伤中微循环障碍的作用及其机制

段洋洋张雅群柴勇张璐萍赵冬梅杨成*

滨州医学院基础医学院人体解剖学教研室，山东烟台264003

收稿日期:2019-12-26 修回日期:2020-02-19 出版日期:2021-02-06 发布日期:2021-02-06
通讯作者: 杨成 E-mail:yangchxn@163.com
基金资助:
山东省自然科学基金;山东省重点研发计划项目

Role and mechanism of stabilizing microtubules of endothelial cells and pericytes in improving the microvasculature dysfunction after spinal cord injury

DUAN Yang-yang ZHANG Ya-qun CHAI Yong ZHANG Lu-ping ZHAO Dong-mei YANG Cheng*

Department of Anatomy, Binzhou Medical University of Basic Medical Sciences, Shandong Yantai264003，China

Received:2019-12-26 Revised:2020-02-19 Online:2021-02-06 Published:2021-02-06
Contact: YANG Cheng E-mail:yangchxn@163.com

摘要/Abstract

摘要：

目的探讨稳定内皮细胞和周细胞微管改善脊髓损伤（SCI）中微循环障碍的作用及其机制。方法培养大鼠脑微血管内皮细胞和周细胞，建立糖氧剥夺（OGD）模型，采用CCK-8法检测细胞活力，采用免疫荧光和Western blotting分别检测α-微管蛋白（α-tubulin）表达；建立脊髓横断损伤模型(n=36)，采用免疫荧光检测大鼠内皮细胞抗原1（RECA-1）及血小板衍生生长因子受体β（PDGFRβ）表达，采用Western blotting检测血管内皮生长因子A（VEGFA）、VEGF受体 2（VEGFR2）、血小板衍生生长因子B（PDGFB）、PDGFRβ、血管生成素1（Ang-1）、特异性酪氨酸激酶受体2（Tie-2）等蛋白表达。结果 OGD组细胞活性显著低于对照组，且具有时间依赖性；埃博霉素B（Epo B）组细胞活性显著高于OGD组；OGD致微管断裂，且随时间延长断裂愈加明显；Epo B组微管比OGD组趋于稳定，断裂减轻。SCI组RECA-1和PDGFRβ表达水平明显低于假手术组，Epo B治疗组RECA-1和PDGFRβ表达水平显著高于SCI组，VEGFA、VEGFR2、PDGFB、PDGFRβ、Ang-1表达水平明显高于SCI组。结论脊髓损伤可导致血管和周细胞减少及微循环障碍，这与细胞微管破坏相关；稳定微管治疗可保护周细胞和微血管，促进微循环重建，建立有利于脊髓损伤修复的微环境。

关键词: 微管, 脊髓损伤, 微循环重建, 内皮细胞, 周细胞, 免疫荧光;大鼠

Abstract:

Objective To explore the role and mechanism of stabilizing microtubules of endothelial cells and pericytes for ameliorating the dysfunction of microvasculature after spinal cord injury（SCI）. Methods The endothelial cells and pericytes from rat brain microvascular tissue (microvessel) were separated and subjected to glucose oxygen deprivation (OGD). The cell viability was detected by CCK-8 and the expression of α-tubulin was detected by immunofluorescence and Western blotting. Rats (n=36) were subjected to dorsal spinal cord transection at T_9-10 vertebra. Expression of rat endothelial cell antigen-1（RECA-1）and platelet-derived growth factor receptor β (PDGFRβ)was localized by immunofluorescence. The expression levels of vascular endothelial growth factor A (VEGFA), VEGF receptor 2 (VEGFR2), platelet-derived growth factor-B (PDGFB), PDGFRβ, angiopoietin-1 (Ang-1) and tyrosine kinase with immunoglobulin-like and epidermal growth factor homology domains（Tie-2）were detected by Western blotting. Results The cell viability in OGD group, which was time-dependent, was significantly lower than that of the control group, whereas the cell viability in OGD and epothilone B（Epo B）group(OGD-Epo B group) was significantly higher than that of the OGD group. Rupture of cell microtubules was observed after OGD, and the breakage of microtubules was time-dependent. However, the microtubules in OGD-Epo B group were more stable and less destroyed than those of the OGD group. The expression levels of RECA-1 and PDGFRβ in the SCI group were significantly lower than those of the sham group, and the levels in SCI-Epo B group were significantly higher than those of the SCI group at day 2 and day 7 post injury. Moreover, the expression levels of VEGFA, VEGFR2, PDGFB, PDGFRβ, Ang-1 in SCI-Epo B group were significantly higher than the levels of the SCI group. Conclusion The endothelial cells and pericytes of microvessels decrease and the microcirculation is dysfunctional after SCI. This corresponds to the microtubule breakage in the cells. The microvessels and pericytes are protected by stabilization treatment of the microtubules, which promotes microcirculation reconstruction and the SCI recovery.

Key words: Microtubule, Spinal cord injury, Microcirculation regeneration, Endothelial cell, Pericyte, Immunofluorescence, Rat

中图分类号:

R744.9

段洋洋张雅群柴勇张璐萍赵冬梅杨成. 稳定内皮和周细胞微管改善脊髓损伤中微循环障碍的作用及其机制[J]. 解剖学报, 2021, 52(1): 21-29.

DUAN Yang-yang ZHANG Ya-qun CHAI Yong ZHANG Lu-ping ZHAO Dong-mei YANG Cheng. Role and mechanism of stabilizing microtubules of endothelial cells and pericytes in improving the microvasculature dysfunction after spinal cord injury[J]. Acta Anatomica Sinica, 2021, 52(1): 21-29.

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稳定内皮和周细胞微管改善脊髓损伤中微循环障碍的作用及其机制

Role and mechanism of stabilizing microtubules of endothelial cells and pericytes in improving the microvasculature dysfunction after spinal cord injury

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