敲除Vanin-1促进肾缺血再灌注损伤后功能恢复的作用及机制

王玲; 汪晓月; 封蕾; 王梨名; 罗佳; 陈佳; 陈客宏

doi:10.16016/j.1000-5404.202009162

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敲除Vanin-1促进肾缺血再灌注损伤后功能恢复的作用及机制

基础医学 | 更新时间：2026-01-09

- 敲除Vanin-1促进肾缺血再灌注损伤后功能恢复的作用及机制
- Effect and mechanism of vanin-1 knockout in promoting recovery of kidney function after renal ischemia reperfusion injury in mice
- 第三军医大学学报 2021年43卷第7期页码：593-598
- 作者机构：
  
  400042 重庆，陆军军医大学（第三军医大学）大坪医院肾内科
- 作者简介：
  
  [陈客宏, E-mail: 19346028@qq.com CHEN Kehong, E-mail: 19346028@qq.com
- 基金信息：
  
  国家自然科学基金面上项目(81770731);重庆市技术创新与应用发展专项面上项目(CSTC2019jscx-msxmX0258)
- DOI：10.16016/j.1000-5404.202009162
  中图分类号： R363.21;R364.12;R692
- 收稿：2020-09-19，
  
  修回：2021-01-11，
  
  纸质出版：2021-04-15
- 稿件说明：
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王玲, 汪晓月, 封蕾, 等. 敲除Vanin-1促进肾缺血再灌注损伤后功能恢复的作用及机制[J]. 第三军医大学学报, 2021,43(7):593-598.

Ling WANG, Xiaoyue WANG, Lei FENG, et al. Effect and mechanism of vanin-1 knockout in promoting recovery of kidney function after renal ischemia reperfusion injury in mice[J]. Journal of Third Military Medical University, 2021, 43(7): 593-598.
王玲, 汪晓月, 封蕾, 等. 敲除Vanin-1促进肾缺血再灌注损伤后功能恢复的作用及机制[J]. 第三军医大学学报, 2021,43(7):593-598. DOI： 10.16016/j.1000-5404.202009162.

Ling WANG, Xiaoyue WANG, Lei FENG, et al. Effect and mechanism of vanin-1 knockout in promoting recovery of kidney function after renal ischemia reperfusion injury in mice[J]. Journal of Third Military Medical University, 2021, 43(7): 593-598. DOI： 10.16016/j.1000-5404.202009162.

摘要

目的

探讨血管非炎症分子1(vascular noninflammatory molecule-1，Vanin-1)在缺血再灌注损伤后肾组织的表达水平及其与肾脏功能恢复的关系。

方法

选取8周龄BALB/c野生型、Vanin-1基因敲除雄鼠，采用完全随机法分为假手术组、缺血再灌注组、敲除组、敲除+缺血再灌注组，每组6只。缺血再灌注组、敲除+缺血再灌注组分离双侧肾蒂后夹闭35 min

后松开夹闭，恢复血流；假手术组、敲除组仅暴露肾蒂，不夹闭。术后0、3、14 d，留取血清、尿液、肾组织标本，检测血清肌酐、尿素氮水平，检测尿液TGF-β水平，PAS染色明确肾组织损伤严重程度，免疫组化检测肾组织Vanin-1表达水平。提取野生型和Vanin-1基因敲除鼠原

代肾小管上皮细胞，传代至第2代

设置对照组、对照+损伤组、敲除组、敲除+损伤组，损伤条件为缺氧（1%O

，94%N

，5%CO

，48 h）、复氧(5%CO

，95%空气，24 h)，模拟体内缺血再灌注损伤模型，收取肾小管上皮细胞和上清，检测Vanin-1蛋白和上清中TGF-β表达水平。

结果

缺血再灌注损伤后第3天，与假手术组相比，缺血再灌注组、敲除+缺血再灌注组血清肌酐、尿素氮水平明显升高（

0.05），PAS染色提示肾组织损伤明显；但缺血再灌注组与敲除+缺血再灌注组小鼠比较，血清肌酐、尿素氮水平无明显差异，PAS染色提示两组肾组织损伤无明显差异。缺血再灌注后第14天，该两组血清肌酐、尿素氮、PAS肾组织损伤均比缺血再灌注后第3天显著恢复，敲除+缺血再灌注组的恢复速度均明显快于缺血再灌注组，PAS肾组织慢性化改变明显轻于缺血再灌注组（

0.05），且敲除+缺血再灌注组尿中TGF-β水平明显低于缺血再灌注组（

0.05）。免疫组化结果提示：损伤后第14天，肾组织Vanin-1表达明显升高（

0.01）

且主要表达于受损肾小管上皮细胞。细胞实验结果显示：予以缺氧-复氧损伤后，对照+损伤组肾小管上皮细胞Vanin-1蛋白表达明显升高(

0.01），上清中TGF-β水平明显高于敲除+损伤组（

0.05）。

结论

Vanin-1可促进缺血再灌注损伤后肾小管上皮细胞TGF-β分泌，延缓肾脏功能恢复，促进急性肾损伤向慢性肾脏病进展。

Abstract

Objective

To explore the expression level of vascular noninflammatory molecule-1 (Vanin-1) in renal tissue after renal ischemia-reperfusion (I/R) injury and its relationship with the recovery of renal function.

Methods

Eight-week-old male BALB/c wild-type and Vanin-1 knockout mice were subjected in this study. They were randomly divided into sham operation group

I/R group

knockout group

knockout+ I/R group

with 6 mice in each group. In the I/R model groups

the bilateral renal pedicles were separated and clamped for 35 min

and then the clamp was released to restore blood flow. For the sham operation and knockout groups

only the renal pedicles were exposed without clipping. Serum

urine

and kidney tissue specimens were collected at 0

and 14 d after surgery to determine blood creatinine and urea nitrogen levels and urine TGF-β level. PAS staining was used to determine the severity of renal tissue damage

and immunohistochemical assay was employed to test the expression of Vanin-1 in renal tissue. After renal tub

ular epithelial cells were isolated from the kidneys of the wild-type and Vanin-1 knockout mice

the cells were primarily cultured and then passed down to the second generation. Then the cells were divided into control group

injury group

knockout group

and knockout+injury group. The cell injury was inflicted with culture condition of hypoxia (1%O

94%N

5%CO

) for 48 h followed by reoxygenation (5%CO

95% air) for 24 h to simulate

in vitro

I/R injury model. The expression of Vanin-1 and supernatant content of TGF-β were measured.

Results

In 3 d after I/R injury

serum creatinine and urea nitrogen levels were significantly higher (

0.05)

and PAS staining indicated obvious renal tissue damage in the I/R and knockout+I/R groups when compared the sham operation group. But there were no significant differences in the levels and the tissue damages between the I/R and knockout+I/R groups. However

on the 14th day after I/R injury

the 2 groups got the serum creatinine and urea nitrogen levels and renal tissue damage significantly recovered than the conditions on the 3rd day

but the knockout+I/R group recovered significantly faster and obtained milder renal damage (

0.05)

lower urine TGF-β level and higher Vanin-1 level when compared with the I/R group. In the

in vitro

study

the mimic I/R injury resulted in obviously increased Vanin-1 level (

0.01)

and increased TGF-β content in the supernatant (

0.05) in the renal tubular epithelial cells when compared in the cells of the knockout+I/R group.

Conclusion

Vanin-1 promotes the secretion of TGF-β in renal tubular epithelial cells after I/R injury

delays the recovery of renal function

and promotes the progression of acute kidney injury (AKI) to chronic kidney disease (CKD).

关键词

Keywords

references

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