Protective mechanism of RNA binding protein LIN28 on diabetic nephropathy
- JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY Vol. 32, Issue 12, Pages: 1048-1053(2023)
- 作者机构：
新疆生产建设兵团医院内分泌科，新疆 乌鲁木齐 830002
- 作者简介：
- 基金信息：
DOI：10.11659/jjssx.11E022103
CLC： R692.9Published：25 December 2023，
Received：18 November 2022，
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王艳丽,李雁,叶芳等.RNA结合蛋白LIN28对糖尿病肾病的保护机制研究[J].局解手术学杂志,2023,32(12):1048-1053.
WANG Yan-li,LI Yan,YE Fang,et al.Protective mechanism of RNA binding protein LIN28 on diabetic nephropathy[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(12):1048-1053.
王艳丽,李雁,叶芳等.RNA结合蛋白LIN28对糖尿病肾病的保护机制研究[J].局解手术学杂志,2023,32(12):1048-1053. DOI： 10.11659/jjssx.11E022103.
WANG Yan-li,LI Yan,YE Fang,et al.Protective mechanism of RNA binding protein LIN28 on diabetic nephropathy[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2023,32(12):1048-1053. DOI： 10.11659/jjssx.11E022103.
摘要
目的
2
探索RNA结合蛋白LIN28对糖尿病肾病（DN）的保护机制。
方法
2
在HEK 293T细胞中通过腺病毒介导过表达或敲低LIN28。通过RNA-Seq转录组测序和生物信息学分析获得过表达LIN28后显著改变的基因和功能信号通路。体外过表达LIN28实验分组为Control组、Treatment组、Adv-LIN28-OE+D-核糖组和Adv-LIN28-NC+D-核糖组；敲低LIN28实验分组为Control组、Treatment组、Adv-shRNA LIN28+D-核糖组和Adv-shRNA NT+D-核糖组。通过ELISA法测定上述各组细胞上清液人晚期糖基化终产物（AGEs）的水平。通过Western blot法测定上述各组细胞内RAGE、NF-κB和MMP-2的表达水平。
结果
2
RNA-Seq转录组测序和GO分析及KEGG分析结果显示，过表达LIN28导致HEK 293T细胞内AGE-RAGE信号通路显著下调（
P
<
0.05）。ELISA结果显示，与Control组相比，Treatment组细胞产生大量AGEs（
P
<
0.05）；与Treatment组相比，Adv-LIN28-OE+D-核糖组细胞AGEs水平明显降低（
P
<
0.05），Adv-shRNA LIN28+D-核糖组细胞AGEs水平显著升高（
P
<
0.05）。Western blot结果显示，与Control组相比，Treatment组细胞内RAGE、NF-κB和MMP-2的表达水平显著升高（
P
<
0.05）；与Treatment组相比，Adv-LIN28-OE+D-核糖组细胞内RAGE、NF-κB和MMP-2的表达水平显著降低（
P
<
0.05），Adv-shRNA LIN28+D-核糖组细胞内RAGE、NF-κB和MMP-2的表达水平则显著升高（
P
<
0.05）。
结论
2
在体外细胞水平上通过腺病毒过表达LIN28，能够导致数千种基因发生显著性差异表达，特别是能够抑制在DN疾病进展中十分关键的糖尿病-并发症相关AGE-RAGE信号通路，发挥保护DN的作用。
Abstract
Objective
2
To explore the protective mechanism of RNA binding protein LIN28 on diabetic nephropathy （DN）.
Methods
2
LIN28 was overexpressed or knocked down by adenovirus in HEK 293T cells. The gene and functional signaling pathway significantly changed after overexpression of LIN28 were obtained through RNA Seq transcriptome sequencing and bioinformatics analysis. HEK 293T cells were divided into the Control group， the Treatment group， the Adv-LIN28-OE+D-ribose group and the Adv-LIN28-NC+D-ribose group in in vitro LIN28 overexpression studies. And HEK 293T cells were divided into the Control group， the Treatment group， the Adv-shRNA LIN28+D-ribose group and the Adv-shRNA NT+D-ribose group in in vitro LIN28 knockdown studies. ELISA was used to detect the levels of human advanced glycation end products （AGEs） of cells supernatants in the above groups. Western blot was used to detect the expression levels of RAGE， NF-κB and MMP-2 of cells in the above groups.
Results
2
The results of RNA-Seq transcriptome sequencing， GO analysis and KEGG analysis showed that overexpression of LIN28 resulted in a significant down-regulation of AGE-RAGE signaling pathway in HEK 293T cells （
P
<
0.05）. The results of ELISA showed that compared with the Control group， the cell in the Treatment group produced a large amount of AGEs （
P
<
0.05）； compared with the Treatment group， the AGEs level of cells in the Adv-LIN28-OE+D-ribose group was significantly decreased （
P
<
0.05）， while the AGEs level of cells in the Adv-shRNA LIN28+D-ribose group was increased（
P
>
0.05）. The results of Western blot showed that compared with the Control group， the expression levels of RAGE， NF-κB and MMP-2 of cells in the Treatment group were significantly increased （
P
<
0.05）； compared with the Treatment group， the expression levels of RAGE， NF-κB and MMP-2 of cells in the Adv-LIN28-OE+D-ribose group were significantly decreased （
P
<
0.05）， while the expression levels of RAGE， NF-κB and MMP-2 of cells in the Adv-shRNA LIN28+D-ribose group were significantly increased（
P
>
0.05）.
Conclusion
2
Overexpression of LIN28 by adenovirus at the cellular level in vitro can lead to significant differential expression of thousands of genes. In particular， it can inhibit the diabetes and complications-related AGE-RAGE signaling pathway， which is critical in the progression of DN disease， and play a role in protecting DN.
关键词
糖尿病肾病LIN28腺病毒AGE-RAGE信号通路NF-κBMMP-2
Keywords
diabetic nephropathyLIN28adenovirusAGE-RAGE signaling pathwayNF-κBMMP-2
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