浏览全部资源
扫码关注微信
VEGF和CD47双修饰外泌体靶向递送治疗沙漠干热环境热射病急性肾损伤的作用及机制研究
Effect and mechanism of targeted delivery of VEGF and CD47 dual-modifiedexosomes in treatment of acute kidney injury induced by heat stroke in rats in the desert dry hot environment.
- 陆军军医大学学报 2024年46卷第18期 页码：2057-2068
- 作者机构：
1.830000 乌鲁木齐，新疆军区总医院急诊医学科
2.400037 重庆, 陆军军医大学第二附属医院生殖医学中心
3.100853 北京，解放军总医院肾内科
4.830000 乌鲁木齐，新疆军区总医院影像科
5.830000 乌鲁木齐，新疆军区总医院重症医学科
6.830000 乌鲁木齐，新疆军区总医院科研教学办公室
- 作者简介：
[杨曦, yxx1984121@163.com Yang Xi, E-mail: yxx1984121@163.com
[张雷, Email: zhanglei_xjjqzyy@163.comZHANG Lei, Email: zhanglei_xjjqzyy@163.com
- 基金信息：2020年度新疆自治区面上项目(2020D01A139)
DOI：10.16016/j.2097-0927.202406020
中图分类号： R594.11;R692.05;R977.6收稿：2024-06-08，
修回：2024-07-20，
纸质出版：2024-09-30
- 稿件说明：
移动端阅览
韩小博, 邓鑫凯, 李红, 等. VEGF和CD47双修饰外泌体靶向递送治疗沙漠干热环境热射病急性肾损伤的作用及机制研究[J]. 陆军军医大学学报, 2024,46(18):2057-2068.
Xiaobo HAN, Xinkai DENG, Hong LI, et al. Effect and mechanism of targeted delivery of VEGF and CD47 dual-modifiedexosomes in treatment of acute kidney injury induced by heat stroke in rats in the desert dry hot environment.[J]. Journal of Army Medical University, 2024, 46(18): 2057-2068.
韩小博, 邓鑫凯, 李红, 等. VEGF和CD47双修饰外泌体靶向递送治疗沙漠干热环境热射病急性肾损伤的作用及机制研究[J]. 陆军军医大学学报, 2024,46(18):2057-2068. DOI： 10.16016/j.2097-0927.202406020.
Xiaobo HAN, Xinkai DENG, Hong LI, et al. Effect and mechanism of targeted delivery of VEGF and CD47 dual-modifiedexosomes in treatment of acute kidney injury induced by heat stroke in rats in the desert dry hot environment.[J]. Journal of Army Medical University, 2024, 46(18): 2057-2068. DOI： 10.16016/j.2097-0927.202406020.
摘要
目的
2
设计一种基于外泌体高效递送的方式，将VEGF和CD47双修饰的外泌体递送到热射病引起的肾损伤，减轻并修复肾损伤。
方法
2
构建靶向肾损伤的融合表达VEGF和CD47的质粒，转染至大鼠骨髓间充质干细胞(bone marrow derived mesenchymal stem cells
BMMSCs)后分离提取外泌体。通过透射电子显微镜及纳米颗粒跟踪分析、蛋白质印迹法等方法对外泌体进行鉴定。经大鼠尾静脉分别注射200 μg DiD标记的未修饰外泌体、VEGF修饰的外泌体和VEGF-CD47双修饰的外泌体，采用小动物活体成像仪检测并分析外泌体对肾脏的靶向作用。60只SD大鼠随机分6组每组10只，sham组为空白对照组(A组，
n
=10)，其余各组均复制热射病肾损伤模型。模型复制成功后12、24、36 h，B组给予相同剂量生理盐水；C组给予质粒空载体(Empty plasmids
Ep)组、D组给予Exos组、E组给予Exos
VEGF
组、F组给予Exos
VEGF-CD47
组。于3次给药治疗后第72小时取肾脏组织和血：从组织水平观察肾组织病理变化并进行损伤评分；检测血清血尿素氮(BUN)、血清肌酐(Scr)评价治疗效果；WB和qRT-PCR分析炎症介质TNF-α、NF-κB的表达水平；检测增殖调控信号分子Ki67、FGF2、pAMPK、pERK
和纤维化调控分子FGF23，综合分析对增殖和抑制纤维化的作用。
结果
2
成功获取了BMMSCs和Exos
VEGF-CD47
并进行鉴定，急性肾损伤模型动物均复制成功。Exos
VEGF-CD47
在活体肾组织中比Exos
Ctrl
组和Exos
VEGF
组的荧光强度高(
P
<
0.05)。治疗后72小时Exos
VEGF-CD47
组，肌酐、尿素氮下降(
P
<
0.000 1)；Exos
VEGF-CD47
组Tubular casts score评分显著低于AKI+Exos组、AKI+Exos
VEGF
组(
P
<
0.000 1)；促炎因子TNF-α、NF-κB蛋白水平明显下调(
P
<
0.000 1)；而Ki67、FGF2显著上调(
P
<
0.05)，FGF23显著下调(
P
<
0.000 1)。
结论
2
VEGF和CD47可以高效靶向热射病急性肾损伤，有效减轻损伤并促进修复，其机制可能与外泌体传递抑制肾组织炎症反应、促进增殖和抑制纤维化有关。
Abstract
Objective
2
To develop an effective method for delivering VEGF and CD47 double-modified exosomes to treat renal damage induced by heat stroke so as to reduce and repair renal damage.
Methods
2
A plasmid fusion-expressing VEGF and CD47 targeting renal injury was constructed
transfected into rat bone marrow derived mesenchymal stem cells (BMMSCs)
and then fusion-exosomes were isolated and extracted. Transmission electron microscopy
nanoparticle tracking analysis
and Western blotting were used to identify the obtained exosomes. Rats were intravenously injected with 200 μg of DiD-labeled unmodified exosomes
VEGF-modified exosomes and VEGF-CD47 double-modified exosomes
respectively
through the tail vein
and the effects of exosomes on the kidneys were detected and analyzed using a small animal
in vivo
imaging instrument. A total of 60 SD rats were randomly divided into 6 groups
with 10 rats in each group
that is
blank control group (group A)
heat stroke-induced renal injury model receiving PBS in 12
24 and 36 h after modelling (group B)
empty plasmid group (group C)
Exos group (group D)
Exos
VEGF
group (group E) and Exos
VEGF-CD47
. Kidney tissue and blood samples were collected in 72 h aft
er 3 times of treatment. Pathological changes in kidney tissue were observed at the tissue level and the damage were scored. Changes in serum blood urea nitrogen (BUN) and serum creatinine (Scr) levels were detected to evaluate the therapeutic effect. Western blotting and qRT-PCR were used to analyze the expression of the pro-inflammatory factors TNF-α and NF-κB
the proliferation regulatory signaling molecules Ki67
FGF2
pAMPK and pERK
and the fibrosis regulatory molecule FGF23
in order to comprehensively analyze the effects on proliferation and inhibition of fibrosis.
Results
2
BMMSCs and Exos
VEGF-CD47
were successfully isolated and characterized
and a rat model of acute kidney injury was effectively constructed. Higher fluorescence intensity was found in the kidney tissue of the Exos
VEGF-CD47
group than the Exos-Ctrl group and Exos
VEGF
group (
P
<
0.05). In 72 h after treatment
the Exos
VEGF-CD47
group had significantly lower serum BUN and Scr levels (
P
<
0.0001)
and notably lower Tubular casts score (
P
<
0.0001)
decreased levels of pro-inflammatory factors TNF-α and NF-κB (
P
<
0.0001)
up-regulated Ki67 and FGF2 expression (
P
<
0.05)
and down-regulated FGF23 expression (
P
<
0.0001) when compared with the AKI+Exos group and AKI+Exos
VEGF
group.
Conclusion
2
VEGF and CD47 show promise in targeting acute kidney injury induced by heat stroke
effectively mitigate damage and facilitate repair
which may be due to exosome-mediated inhibition of renal tissue inflammation
promotion of proliferation
and inhibition of fibrosis.
关键词
Keywords
references
何嘉骐, 倪军, 张静. 热射病相关肾脏损伤的机制及研究进展[J]. 中华灾害救援医学, 2021, 9(1): 754-757, 767. DOI: 10.13919/j.issn.2095-6274.2021.12.008.
HE J Q, NI J, ZHANG J. Review on the mechanism and research progress ofrenal injury related heat radiation disease[J]. Chin J Disaster Med, 2021, 9(1): 754-757, 767. DOI: 10.13919/j.issn.2095-6274.2021.12.008.
BOUCHAMA A, ABUYASSIN B, LEHE C, et al . Classic and exertional heatstroke[J ] . Nat Rev Dis Primers, 2022, 8(1): 8. DOI: 10.1038/s41572-021-00334-6.
SHEN C F, WEI D H, WANG G J, et al . Swine hemorrhagic shock model and pathophysiological changes ina desert dry-heat environment[J ] . PLoS One, 2021, 16(1): e0244727. DOI: 10.1371/journal.pone.0244727.
WEN F L, XU Y J, XUE L E , et al . Proteomics analyses of acute kidney injury biomarkers in a rat exertional heat stroke model[J ] . Front Physiol, 2023, 14: 1176998. DOI: 10.3389/fphys.2023.1176998.
WU M, WANG C L, LIU Z Y, et al . Clinical characteristics and risk factors associated with acute kidney injury inpatient with exertional heatstroke: an over 10-year intensive care survey[J ] . Front Med, 2021, 8: 678434. DOI: 10.3389/fmed.2021.678434.
WITWER K W, THÉRY C. Extracellular vesicles or exosomes? On primacy, precision, and popularity influencing a choice of nomenclature[J]. J Extracell Vesicles, 2019, 8(1): 1648167. DOI: 10.1080/20013078.2019.1648167.
BOST J P, SAHER O, HAGEY D, et al . Growth media conditions influence the secretion route and release levels of engineered extracellular vesicles[J ] . Adv Healthc Mater, 2022, 11(5): e2101658. DOI: 10.1002/adhm.202101658.
OH S, KWON S H. Extracellular vesicles in acute kidney injury and clinical applications[J]. Int J Mol Sci, 2021, 22(16): 8913. DOI: 10.3390/ijms22168913.
KUSABA T, LALLI M, KRAMANN R, et al . Differentiated kidney epith elial cells repair injured proximal tubule[J ] . Proc Natl Acad Sci U S A, 2014, 111(4): 1527-1532. DOI: 10.1073/pnas.1310653110.
ZHA Y, LI Y W, LIN T Y, et al . Progenitor cell-derived exosomes endowed with VEGF plasmids enhance osteogenic induction and vascular remodeling in large segmental bone defects[J ] . Theranostics, 2021, 11(1): 397-409. DOI: 10.7150/thno.50741.
JOSUTTIS D, SCHWEDLER C, HEYMANN G, et al . Vascular endothelial growth factor as potential biomarker for COVID-19 severity[J ] . J Intensive Care Med, 2023, 38(12): 1165-1173. DOI: 10.1177/08850666231186787.
LIU C, WANG J, HU J, et al . Extracellular vesicles for acute kidney injury in preclinical rodent models: a meta-analysis[J ] . Stem Cell Res Ther, 2020, 11(1): 11. DOI: 10.1186/s13287-019-1530-4.
WANG J, DONG Y, LI Y, et al . Designer Exosomes for active targeted chemo-photothermal synergistic tumor therapy[J ] . Adv Funct Mater, 2018;28(18): 1-14. DOI: 10.1002/adfm.201707360.
KANELLIS J, MUDGE S J, FRASER S, et al . Redistribution of cytoplasmic VEGF to the basolateral aspe ct of renal tubular cells in ischemia-reperfusion injury[J ] . Kidney Int, 2000, 57(6): 2445-2456. DOI: 10.1046/j.1523-1755.2000.00103.x.
ZHONG X, TANG T T, SHEN A R, et al . Tubular epithelial cells-derived small extracellular vesicle-VEGF-A promotes peritubular capillary repair in ischemic kidney injury[J ] . NPJ Regen Med, 2022, 7(1): 73. DOI: 10.1038/s41536-022-00268-x.
PICKERING J W, ENDRE Z H. Bench to bedside: the next steps for biomarkers in acute kidney injury[J]. Am J Physiol Renal Physiol, 2016, 311(4): F717-F721. DOI: 10.1152/ajprenal.00268.2016.
BELHADJ Z, HE B, DENG H L, et al . A combined "eat me/don't eat me" strategy based on extracellular vesicles for anticancer nanomedicine[J ] . J Extracell Vesicles, 2020, 9(1): 1806444. DOI: 10.1080/20013078.2020.1806444.
ZHAO H, SONG S S, MA J W, et al . CD47 as a promising therapeutic target in oncology[J ] . Front Immunol, 2022, 13: 757480. DOI: 10.3389/fimmu.2022.757480.
ZHOU R O, LIU J W, ZHANG D, et al . Heatstroke model for desert dry-heat environment and observed organ damage[J ] . Am J Emerg Med, 20 14, 32(6): 573-579. DOI: 10.1016/j.ajem.2014.02.017.
ZHAO Y H, SHEN C F, WANG G J, et al . Curcumin alleviates acute kidney injury in a dry-heat environment by reducing oxidative stress and inflammation in a rat model[J ] . J Biochem Mol Toxicol, 2021, 35(1): e22630. DOI: 10.1002/jbt.22630.
RICCI Z, ROMAGNOLI S. Acute kidney injury: diagnosis and classification in adults and children[J]. Contrib Nephrol, 2018, 193: 1-12. DOI: 10.1159/000484956.
GOTO H, NAKASHIMA M, NAKASHIMA H, et al . Heat acclimation ameliorated heat stress-induced acute kidney injury and prevented changes in kidney macrophages and fibrosis[J ] . Am J Physiol Renal Physiol, 2022, 323(3): F243-F254. DOI: 10.1152/ajprenal.00065.2022.
HADE M D, SUIRE C N, SUO Z C. Mesenchymal stem cell-derived exosomes: applications in regenerative medicine[J]. Cells, 2021, 10(8): 1959. DOI: 10.3390/cells10081959.
LIU C, CHI K, GENG X D, et al . Exogenous biological renal support improves kidney function in mice with rhabdomyolysis-induced acute kidney injury[J ] . Front Med, 2021, 8: 655787. DOI: 10.3389/fmed.2021.655787.
XUE L, GUO W L, LI L, et al . Metabolomic profiling identifies a novel mechanism for heat stroke-related acute kidney injury[J ] . Mol Med Rep, 2021, 23(4): 241. DOI: 10.3892/mmr.2021.11880.
全军热射病防治专家组, 热射病急诊诊断与治疗专家共识组. 热射病急诊诊断与治疗专家共识(2021版)[J]. 中华急诊医学杂志, 2021, 30(11): 1290-1299. DOI: 10.3760/cma.j.issn.1671-0282.2021.11.002.
Army Expert Group on Prevention and Treatment of Heat Stroke Disease, the consensus group of experts on Emergency Diagnosis and Treatment of Heat Stroke. Expert consensus on emergency diagnosis and treatment of heat stroke (2021 edition)[J]. Chin J Emerg Med, 2021, 30(11): 1290-1299. DOI: 10.3760/cma.j.issn.1671-0282.2021.11.002.
LI Y, ZHU X T, ZHANG M, et al . Heatstroke-induced hepatocyte exosomes promote liver injury by activating the NOD-like receptor signaling pathway in mice[J ] . PeerJ, 2019, 7: e8216. DOI: 10.7717/peerj.8216.
HUN M, WEN H, HAN P, et al . Bibliometric analysis of scientific papers on extracellular vesicles in kidney disease published between 1999 and 2022[J ] . Front Cell Dev Biol, 2022, 10: 1070516. DOI: 10.3389/fcell.2022.1070516.
CHEN Y M, ZHANG C Y, DU Y J, et al . Exosomal transfer of microRNA-590-3p between renal tubular epithelial cells after renal ischemia-reperfusion injury regulates autophagy by targeting TRAF6[J ] . Chin Med J, 2022,135(20): 2467-2477. DOI: 10.1097/CM9.0000000000002377.
SALUNKHE S, DHEERAJ, BASAK M, et al . Surface functionalization of exosomes for target-specific delivery and in vivo imaging & tracking: Strategies and significance[J ] . J Control Release, 2020, 326: 599-614. DOI: 10.1016/j.jconrel.2020.07.042.
KLABUKOV I, BALYASIN M, KRASILNIKOVA O, et al . Angiogenic modification of microfibrous polycaprolactone by pCMV-VEGF165 plasmid promotes local vascular growth after implantation in rats[J ] . Int J Mol Sci, 2023, 24(2): 1399. DOI: 10.3390/ijms24021399.
WYSOCZYNSKI M, PATHAN A, MOORE J B 4th, et al . Pro-angiogenic actions of CMC-derived extracellular vesicles rely on selective packaging of angiopoietin 1 and 2, but not FGF-2 and VEGF[J ] . Stem Cell Rev Rep, 2019, 15(4): 530-542. DOI: 10.1007/s12015-019-09891-6.
MAISTO R, OLTRA M, VIDAL-GIL L, et al . ARPE-19-derived VEGF-containing exosomes promote neovascularization in HUVEC: the role of the melanocortin receptor 5[J ] . Cell Cycle, 2019, 18(4): 413-424. DOI: 10.1080/15384101.2019.1568745.
SIMONS M, GORDON E, CLAESSON-WELSH L. Mechanisms and regulation of endothelial VEGF receptor signalling[J]. Nat Rev Mol Cell Biol, 2016, 17(10): 611-625. DOI: 10.1038/nrm.2016.87.
KIM H, YUN N R, MUN D, et al . Cardiac-specific delivery by cardiac tissue-targeting peptide-expressing exosomes[J ] . Biochem Biophys Res Commun, 2018, 499(4): 803-808. DOI: 10.1016/j.bbrc.2018.03.227.
MURPHY D E, DE JONG O G, BROUWER M, et al . Extracellular vesicle-based therapeutics: natural versus engineered targeting and trafficking[J ] . Exp Mol Med, 2019, 51(3): 1-12. DOI: 10.1038/s12276-019-0223-5.
YUAN F F, CAI J Z, WU J F, et al . Z-DNA binding protein 1 promotes heatstroke-induced cell death[J ] . Science, 2022, 376(6593): 609-615. DOI: 10.1126/science.abg5251.
FIGUREK A, RROJI M, SPASOVSKI G. FGF23 in chronic kidney disease: bridging the heart and Anemia[J]. Cells, 2023, 12(4): 609. DOI: 10.3390/cells12040609.
0
浏览量
150
下载量
0
CSCD
- 网络PDF下载
- Meta-XML下载
- BibTeX下载
- Endnote下载
- RefMan 下载
- NoteExpress下载
- NoteFirst下载
关联资源
相关文章
相关作者
相关机构
- 技术支持由北京北大方正电子有限公司提供 京ICP备09064830号-19
京公网安备11010802024621 - 本系统建议在Chrome、 IE9+ 以上版本浏览器阅读本站内容，360浏览器请切换至极速模式
- Cookies帮助我们提供服务并提供个性化体验。使用本网站，即表示您同意我们使用Cookies
