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.

Xiaobo HAN; Xinkai DENG; Hong LI; Chao LIU; Jiawei GAO; Xiao LI; Yahao CHAI; Xiaoli HAN; Hongwei HAN; Yinjie ZHAO; Xi Yang; Lei ZHANG

doi:10.16016/j.2097-0927.202406020

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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.

Basic Medicine | 更新时间：2026-01-09

- 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.
- Journal of Army Medical University Vol. 46, Issue 18, Pages: 2057-2068(2024)
- 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：
- 基金信息：
  
  the General Project of Science Foundation of Xinjiang Uygur Autonomous Region(2020D01A139)
- DOI：10.16016/j.2097-0927.202406020
  CLC： R594.11;R692.05;R977.6
- Received：08 June 2024，
  
  Revised：2024-07-20，
  
  Published：30 September 2024
- 稿件说明：
移动端阅览
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：

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

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

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，

=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

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(

0.05)。治疗后72小时Exos

VEGF-CD47

组，肌酐、尿素氮下降(

0.000 1)；Exos

VEGF-CD47

组Tubular casts score评分显著低于AKI+Exos组、AKI+Exos

VEGF

组(

0.000 1)；促炎因子TNF-α、NF-κB蛋白水平明显下调(

0.000 1)；而Ki67、FGF2显著上调(

0.05)，FGF23显著下调(

0.000 1)。

结论

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

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

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

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 (

0.05). In 72 h after treatment

the Exos

VEGF-CD47

group had significantly lower serum BUN and Scr levels (

0.0001)

and notably lower Tubular casts score (

0.0001)

decreased levels of pro-inflammatory factors TNF-α and NF-κB (

0.0001)

up-regulated Ki67 and FGF2 expression (

0.05)

and down-regulated FGF23 expression (

0.0001) when compared with the AKI+Exos group and AKI+Exos

VEGF

group.

Conclusion

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

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