内皮祖细胞CXC趋化因子受体7对脑缺血再灌注后血管新生的作用

doi:10.3969/j.issn.1006-9771.2019.11.007

《中国康复理论与实践》 ›› 2019, Vol. 25 ›› Issue (11): 1283-1292.doi: 10.3969/j.issn.1006-9771.2019.11.007

内皮祖细胞CXC趋化因子受体7对脑缺血再灌注后血管新生的作用

范加维¹, 杨拯¹, 王近吴¹, 范道贵², 蒋仕秋³, 曾江华⁴, 易红梅³, 戴小珍², 刘海荣³

1.成都医学院基础医学院，四川成都市 610500
2.成都医学院生物科学与技术学院，四川成都市 610500
3.成都医学院第一附属医院，四川成都市 610500
4.遂宁市第一人民医院，四川遂宁市 629000

收稿日期:2019-06-30 修回日期:2019-08-26 出版日期:2019-11-25 发布日期:2019-11-21
通讯作者: 刘海荣，E-mail: 1332801661@qq.com E-mail:1332801661@qq.com
作者简介:范加维(1993-)，男，汉族，重庆市人，硕士研究生，主要研究方向：心血管病变及干预机制研究。
基金资助:
1.国家自然科学基金项目(No. 81770305)；2.四川省教育厅资助科研项目(No. 18ZA0144)；3.四川省科技创新苗子工程培育项目(No. 2018071)；4.四川省医学会项目(No. S17021)；5.四川省教育厅科研重点项目(No. 18ZB0168)；6.成都医学院创新团队项目(No. CYTD17-01)

Effects of CXC-chemokine Receptor 7 Expression in Endothelial Progenitor Cells on Angiogenesis after Cerebral Ischemia-reperfusion Injury

FAN Jia-wei¹, YANG Zheng¹, WANG Jin-wu¹, FAN Dao-gui², JIANG Shi-qiu³, ZENG Jiang-hua⁴, YI Hong-mei³, DAI Xiao-zhen², LIU Hai-rong³

1.School of Basic Medicine, Chengdu Medical College, Chengdu, Sichuan 610500, China
2.School of Biosciences and Technology, Chengdu Medical College, Chengdu, Sichuan 610500, China
3.The First Affiliated Hospital of Chengdu Medical College, Chengdu, Sichuan 610500, China
4.Suining First People's Hospital, Suining, Sichuan 629000, China

Received:2019-06-30 Revised:2019-08-26 Published:2019-11-25 Online:2019-11-21
Contact: LIU Hai-rong, E-mail: 1332801661@qq.com E-mail:1332801661@qq.com
Supported by:
Supported by National Natural Science Foundation of China (No. 81770305), Sichuan Education Department Scientific Research Fund (No. 18ZA0144), Sichuan Sci-tech Innovation Seedling Project Cultivation Fund (No. 2018071), Sichuan Medical Association Project (No. S17021), Sichuan Education Department Key Scientific Research Fund (No. 18ZB0168) and Chengdu Medical College Innovative Team Project (No. CYTD17-01)

摘要/Abstract

摘要： 目的探讨上调内皮祖细胞(EPCs)中CXC趋化因子受体7 (CXCR7)的表达对EPCs促进脑缺血再灌注后血管新生的作用。方法从人脐带血分离培养EPCs并鉴定。构建CXCR7过表达慢病毒载体，转染EPCs，实时定量PCR和Western blotting检测转染效率。体外通过管样结构形成实验和Annexin V/PI染色分别检测氧化低密度脂蛋白(ox-LDL)处理下EPCs管样结构形成及抗凋亡能力。线栓法制备大鼠脑缺血再灌注损伤模型，再灌注24 h后根据神经功能评分，将合格模型随机分为三组，PBS组(n = 12)尾静脉注射磷酸盐缓冲液，对照组(n = 12)注射对照病毒转染EPCs，移植组(n = 12)注射CXCR7过表达病毒转染EPCs。分别于干预后7 d和14 d予神经功能评分，14 d测定大鼠脑梗死体积，冰冻切片观察缺血部位GFP阳性细胞数量和毛细血管密度。结果转染CXCR7过表达载体能明显上调EPCs中CXCR7表达(P < 0.01)；CXCR7表达上调后，EPCs在ox-LDL处理下管样结构形成能力改善(P < 0.05)，EPCs凋亡减轻(P < 0.05)。相比PBS组和对照组，移植组神经功能改善(P < 0.05)，梗死体积减少(P < 0.05)，缺血部位EPCs数量和毛细血管密度增加(P < 0.05)。结论上调CXCR7可改善EPCs的存活和血管形成功能，促进血管新生，改善脑缺血再灌注损伤修复。

关键词: 脑缺血再灌注, 内皮祖细胞, CXC趋化因子受体7, 血管新生, 大鼠

Abstract: Objective To explore the effect of upregulating CXC-chemokine receptor 7 (CXCR7) in endothelial progenitor cells (EPCs) on angiogenesis after cerebral ischemia-reperfusion injury. Methods EPCs were isolated and cultured from human umbilical cord blood and identified. Then, the EPCs were transfected with CXCR7 overexpression lentiviral vector, and the expression of CXCR7 was identified with real-time PCR and Western blotting. The tube-like structure formation and apoptosis of EPCs under oxidized low density lipoprotein (ox-LDL) were detected with tube-like structure formation test and Annexin V/PI staining. Cerebral ischemia-reperfusion injury model in rats was established, and the qualified model rats were randomly divided into three groups after 24 hours reperfusion: PBS group (n = 12) was injected with phosphate buffers through tail vein, control group (n = 12) was injected the EPCs infected with control lentiviral vector, and CXCR7 group (n = 12) was injected with EPCs infected with CXCR7 overexpression lentiviral vector. Neurological function scores were determined seven and 14 days after transplantation. The cerebral infarct volume was measured, the number of GFP-positive cells in the ischemic site and the density of capillary were observed. Results The expression of CXCR7 in EPCs increased after transfection (P < 0.01). Overexpression of CXCR7 improved tube formation and reduced apoptosis of EPCs under ox-LDL (P < 0.05). Compared with PBS and control groups , neurological function improved in CXCR7 group, with less infarct volume, more GFP-positive cells and density of capillary (P < 0.05). Conclusion Up-regulating CXCR7 can improve the survival and angiogenesis of EPCs, and improve the repair of cerebral ischemia-reperfusion injury.

Key words: cerebral ischemia-reperfusion, endothelial progenitor cells, CXC-chemokine receptor 7, angiogenesis, rats

中图分类号:

R743.3

范加维, 杨拯, 王近吴, 范道贵, 蒋仕秋, 曾江华, 易红梅, 戴小珍, 刘海荣. 内皮祖细胞CXC趋化因子受体7对脑缺血再灌注后血管新生的作用[J]. 《中国康复理论与实践》, 2019, 25(11): 1283-1292.

FAN Jia-wei, YANG Zheng, WANG Jin-wu, FAN Dao-gui, JIANG Shi-qiu, ZENG Jiang-hua, YI Hong-mei, DAI Xiao-zhen, LIU Hai-rong. Effects of CXC-chemokine Receptor 7 Expression in Endothelial Progenitor Cells on Angiogenesis after Cerebral Ischemia-reperfusion Injury[J]. 《Chinese Journal of Rehabilitation Theory and Practice》, 2019, 25(11): 1283-1292.

参考文献

1 LiuR, YangS H. Window of opportunity:estrogen as a treatment for ischemic stroke [J]. Brain Res, 2013, 1514: 83-90.
2 MohammadA, GhaniU, SchwindtB, et al. Endothelial progenitor cells in patients with acute cerebrovascular ischemia [J]. Can J Neurol Sci, 2010, 37(6): 797-802.
3 ZhangJ, ChenC, HuB, et al. Exosomes derived from human endothelial progenitor cells accelerate cutaneous wound healing by promoting angiogenesis through Erk1/2 signaling [J]. Int J Biol Sci, 2016, 12(12): 1472-1487.
4 XuQ B. Endothelial progenitor cells in angiogenesis [J]. Sheng Li Xue Bao, 2005, 57(1): 1-6.
5 XinB, LiuC L, YangH, et al. Prolonged fasting improves endothelial progenitor cell-mediated ischemic angiogenesis in mice [J]. Cell Physiol Biochem, 2016, 40(3-4): 693-706.
6 ChungC P, HuangP H, ChenJ S, et al. The level of circulating endothelial progenitor cell is associated with cerebral vasoreactivity: a pilot study [J]. Biomed Res Int, 2015, 2015: 258279.
7 DaiX, YanX, ZengJ, et al. Elevating CXCR7 improves angiogenic function of EPCs via Akt/GSK-3beta/Fyn-Mediated Nrf2 activation in diabetic limb ischemia [J]. Circul Res, 2017, 120(5): e7-e23.
8 SobrinoT, HurtadoO, MoroM A, et al. The increase of circulating endothelial progenitor cells after acute ischemic stroke is associated with good outcome [J]. Stroke, 2007, 38(10): 2759-2764.
9 FadiniG P, AgostiniC, AvogaroA. Endothelial progenitor cells in cerebrovascular disease [J]. Stroke, 2005, 36(6): 1112-1113.
10 BoytsovS, LogunovaN, KhomitskayaY. CEPHEUS II investigators. Suboptimal control of lipid levels: results from the non-interventional Centralized Pan-Russian Survey of the Undertreatment of Hypercholesterolemia II (CEPHEUS II) [J]. Cardiovasc Diabetol, 2017, 16(1): 158.
11 BravataD M, DaggyJ, BroschJ, et al. Comparison of risk factor control in the year after discharge for ischemic stroke versus acute myocardial infarction [J]. Stroke, 2018, 49(2): 296-303.
12 FanY, ShenF, FrenzelT, et al. Endothelial progenitor cell transplantation improves long-term stroke outcome in mice [J]. Ann Neurol, 2010, 67: 488-497.
13 RouhlR P, van OostenbruggeR J, DamoiseauxJ, et al. Endothelial progenitor cell research in stroke: a potential shift in patho-physiological and therapeutical concepts [J]. Stroke, 2008, 39: 2158-2165.
14 GuyonA. CXCL12 chemokine and its receptors as major players in the interactions between immune and nervous systems [J]. Front Cell Neurosci, 2014, 8: 65.
15 ZhangY, ZhangH, LinS, et al. SDF-1/CXCR7 chemokine signaling is induced in the peri-infarct regions in patients with ischemic stroke [J]. Aging Dis, 2018, 9: 287-295.
16 ChengX, WangH, ZhangX, et al. The role of SDF-1/CXCR4/CXCR7 in neuronal regeneration after cerebral ischemia [J]. Front Neurosci, 2017, 11: 590.
17 LiY, TangG, LiuY, et al. CXCL12 gene therapy ameliorates ischemia-induced white matter injury in mouse brain [J]. Stem Cells Transl Med, 2015, 4: 1122-1130.
18 DaiX, TanY, CaiS, et al. The role of CXCR7 on the adhesion, proliferation and angiogenesis of endothelial progenitor cells [J]. J Cell Mol Med, 2011, 15: 1299-1309.
19 O'NeillK M, CampbellD C, EdgarK S, et al. NOX4 is a major regulator of cord blood-derived endothelial colony-forming cells which promotes postischaemic revascularization [J]. Heart2018, 104(Suppl 4): A1-A10.
20 ZhangH, TaoY, RenS, et al. Simultaneous harvesting of endothelial progenitor cells and mesenchymal stem cells from the human umbilical cord [J]. Exp Ther Med, 2018, 15: 806-812.
21 SmithP J, HowesE A, TreherneJ E. Cell proliferation in the repairing adult insect central nervous system: incorporation of the thymidine analogue 5-bromo-2-deoxyuridine in vivo [J]. J Cell Sci, 1990, 95(Pt 4): 599-604.
22 HuC H, LiZ M, DUZ M, et al. Human umbilical cord-derived endothelial progenitor cells promote growth cytokines-mediated neorevascularization in rat myocardial infarction [J].Chin Med J, 2009, 122(5): 548-555.
23 范加维,杨森,杨拯,等. 原花青素对脑缺血再灌注损伤后肠道功能的保护作用[J]. 中国康复理论与实践, 2015, 21(10): 1138-1144.
24 ZhangY, LiY W, WangY X, et al. Remifentanil preconditioning alleviating brain damage of cerebral ischemia reperfusion rats by regulating the JNK signal pathway and TNF-α/TNFR1 signal pathway [J]. Mol Biol Rep, 2013, 40(12): 6997-7006.
25 BalajiS, KingA, CrombleholmeT M, et al. The role of endothelial progenitor cells in postnatal vasculogenesis: implications for therapeutic neovascularization and wound healing [J]. Adv Wound Care (New Rochelle), 2013, 2(6): 283-295.
26 AokiM, YasutakeM, MuroharaT. Derivation of functional endothelial progenitor cells from human umbilical cord blood mononuclear cells isolated by a novel cell filtration device [J]. Stem Cells, 2004, 22(6): 994-1002.
27 UrbichC, DimmelerS. Endothelial progenitor cells: characterization and role in vascular biology [J]. Circ Res, 2004, 95(4): 343-353.
28 QiuJ, LiW, FengS, et al. Transplantation of bone marrow-derived endothelial progenitor cells attenuates cerebral ischemia and reperfusion injury by inhibiting neuronal apoptosis, oxidative stress and nuclear factor-κB expression [J]. Int J Mol Med, 2013, 31(1): 91-98.
29 BaiY Y, PengX G, WangL S, et al. Bone marrow endothelial progenitor cell transplantation after ischemic stroke: an investigation into its possible mechanism [J]. CNS Neurosci Ther, 2015, 21(11): 877-886.

[1]	李芳, 霍速, 杜巨豹, 刘秀贞, 李小爽, 宋为群. 经颅直流电刺激联合任务导向性康复训练对脊髓损伤大鼠前肢运动障碍的效果[J]. 《中国康复理论与实践》, 2023, 29(7): 777-781.
[2]	康晓宇, 刘丽旭, 王文竹, 王云雷. 普拉克索联合左旋多巴对全脑缺血再灌注损伤大鼠认知能力及线粒体功能的影响[J]. 《中国康复理论与实践》, 2023, 29(5): 533-540.
[3]	罗兰, 李璐, 金沐. 氙气后处理对脊髓缺血再灌注损伤的效果：Akt信号通路和自噬机制[J]. 《中国康复理论与实践》, 2023, 29(2): 174-181.
[4]	黄志霖, 徐发邵, 施静, 黄淦, 刘梅芳, 张霞辉. 线栓法建立卒中后吞咽障碍的大鼠模型[J]. 《中国康复理论与实践》, 2023, 29(10): 1147-1153.
[5]	秦彦强, 董浩, 孙迎春, 程先宽, 姚海江. 不同针刺方案对卒中后抑郁大鼠神经递质及相关炎性因子的影响[J]. 《中国康复理论与实践》, 2023, 29(1): 30-37.
[6]	缪培,张通,米海霞,张伟东. 不同线栓法复制局灶性脑缺血模型大鼠恢复期学习记忆能力的差异及其机制[J]. 《中国康复理论与实践》, 2022, 28(7): 789-796.
[7]	周小珏,冯婧,庞日朝,刘捷,张安仁. 隔日限食减轻脊髓损伤大鼠炎症反应的芳香烃受体/细胞因子信号传导抑制因子2/核转录因子-κB信号通路机制[J]. 《中国康复理论与实践》, 2022, 28(5): 544-551.
[8]	宋绍霏,侯园园,王云雷,张通. 异常光周期所致昼夜节律紊乱对大鼠腓肠肌时钟基因和葡萄糖摄取相关基因表达节律的影响[J]. 《中国康复理论与实践》, 2022, 28(5): 552-558.
[9]	李童,方志鹏,邵玉萍,王平. 有氧运动对睡眠剥夺大鼠学习记忆及海马神经元突触可塑性的效果[J]. 《中国康复理论与实践》, 2022, 28(11): 1270-1277.
[10]	王静怡,尹杰,刘建成,庞日朝,王文春. 蜘蛛香环烯醚萜类成分对急性脊髓损伤大鼠神经细胞焦亡的影响[J]. 《中国康复理论与实践》, 2021, 27(6): 653-660.
[11]	周文美,陶陶,吴霜,王廷龙,杨正奕,张莹. 丰富环境对脑缺血再灌注损伤大鼠神经功能和缺血半暗带区葡萄糖代谢的影响[J]. 《中国康复理论与实践》, 2021, 27(5): 522-529.
[12]	王琼芬,王风波,王科,钟永强,王娇娇. 电针风池对急性脑梗死大鼠脑星形胶质细胞和神经元的保护效果[J]. 《中国康复理论与实践》, 2021, 27(3): 302-309.
[13]	李向哲,丁洁,王庆华,董传明,王彤,吴勤峰. 减重平板训练对脊髓损伤大鼠神经病理性疼痛及脊髓后角谷氨酸脱羧酶-65/67表达的影响[J]. 《中国康复理论与实践》, 2021, 27(2): 131-136.
[14]	艾坤,许明,刘琼,邓石峰,刘继生,祁芳,易细芹,瞿启睿,张泓. 电针对骶上脊髓损伤后逼尿肌亢进大鼠逼尿肌中环腺苷酸和蛋白激酶A含量及肌球蛋白轻链激酶磷酸化的影响[J]. 《中国康复理论与实践》, 2021, 27(2): 137-144.
[15]	凌梦钰,杨一卓,刘帅,叶超群. 运动训练对脑缺血再灌注大鼠认知功能及前额皮层神经元核抗原和突触素1表达的效果[J]. 《中国康复理论与实践》, 2021, 27(11): 1272-1281.

内皮祖细胞CXC趋化因子受体7对脑缺血再灌注后血管新生的作用

Effects of CXC-chemokine Receptor 7 Expression in Endothelial Progenitor Cells on Angiogenesis after Cerebral Ischemia-reperfusion Injury

PDF (PC)

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

Metrics

本文评价

推荐阅读 0