Basic Study
Copyright ©The Author(s) 2021. Published by Baishideng Publishing Group Inc. All rights reserved.
World J Diabetes. Jul 15, 2021; 12(7): 1116-1130
Published online Jul 15, 2021. doi: 10.4239/wjd.v12.i7.1116
Expression and role of P-element-induced wimpy testis-interacting RNA in diabetic-retinopathy in mice
Yong Yu, Kai-Ming Ren, Xiao-Long Chen
Yong Yu, Xiao-Long Chen, Department of Ophthalmology, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
Kai-Ming Ren, Department of Thoracic Surgery, Shengjing Hospital of China Medical University, Shenyang 110004, Liaoning Province, China
Author contributions: Yu Y and Ren KM designed this study, conducted the experimental analysis, and wrote the manuscript; Chen XL revised the manuscript; all authors have read and approved the final manuscript.
Supported by National Natural Science Foundation of China, No. 81570866.
Institutional animal care and use committee statement: The study was reviewed and approved by the Institutional Review Board of Shengjing Hospital of China Medical University (Approval No. 2020PS078K).
Conflict-of-interest statement: No potential conflicts of interest relevant to this article are reported.
Data sharing statement: No additional data are available.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See: http://creativecommons.org/Licenses/by-nc/4.0/
Corresponding author: Xiao-Long Chen, MD, Chief Doctor, Surgeon, Department of Ophthalmology, Shengjing Hospital of China Medical University, No. 39 Huaxiang Road, Shenyang 110004, Liaoning Province, China. chenxl@sj-hospital.org
Received: March 2, 2021
Peer-review started: March 2, 2021
First decision: April 6, 2021
Revised: April 11, 2021
Accepted: May 20, 2021
Article in press: May 20, 2021
Published online: July 15, 2021
Processing time: 132 Days and 1.6 Hours
Abstract
BACKGROUND

As one of the major microvascular complications of diabetes, diabetic retinopathy (DR) is the leading cause of blindness in the working age population. Because the extremely complex pathogenesis of DR has not been fully clarified, the occurrence and development of DR is closely related to tissue ischemia and hypoxia and neovascularization The formation of retinal neovascularization (RNV) has great harm to the visual acuity of patients.

AIM

To investigate the expression of P-element-induced wimpy testis-interacting RNA (piRNA) in proliferative DR mice and select piRNA related to RNV.

METHODS

One hundred healthy C57BL/6J mice were randomly divided into a normal group as control group (CG) and proliferative DR (PDR) group as experimental group (EG), with 50 mice in each group. Samples were collected from both groups at the same time, and the lesions of mice were evaluated by hematoxylin and eosin staining and retinal blood vessel staining. The retinal tissues were collected for second-generation high-throughput sequencing, and the differentially expressed piRNA between the CG and EG was detected, and polymerase chain reaction (PCR) was conducted for verification. The differentially obtained piRNA target genes and expression profiles were enrichment analysis based on gene annotation (Gene Ontology) and Kyoto Encyclopedia of Genes and Genomes.

RESULTS

In the CG there was no perfusion area, neovascularization and endothelial nucleus broke through the inner boundary membrane of retinap. In the EG, there were a lot of nonperfused areas, new blood vessels and endothelial nuclei breaking through the inner boundary membrane of the retina. There was a statistically significant difference in the number of vascular endothelial nuclei breaking through the inner retinal membrane between the two groups. High-throughput sequencing analysis showed that compared with the CG, a total of 79 piRNAs were differentially expressed in EG, among which 43 piRNAs were up-regulated and 36 piRNAs were down-regulated. Bioinformatics analysis showed that the differentially expressed piRNAs were mainly concentrated in the signaling pathways of angiogenesis and cell proliferation. Ten piRNAs were selected for PCR, and the results showed that the expression of piR-MMU-40373735, piR-MMU-61121420, piR-MMU-55687822, piR-MMU-1373887 were high, and the expression of piR-MMU-7401535, piR-MMU-4773779, piR-MMU-1304999, and piR-MMU-5160126 were low, which were consistent with the sequencing results.

CONCLUSION

In the EG, the abnormal expression of piRNA is involved in the pathway of angiogenesis and cell proliferation, suggesting that piRNAs have some regulatory function in proliferative diabetic-retinopathy.

Keywords: P-element-induced wimpy testis-interacting RNA; P-element-induced wimpy testis protein; High-throughput sequencing; Neovascularization; Diabetic retinopathy in mice; Bioinformatics

Core Tip: As one of the major microvascular complications of diabetes, diabetic retinopathy is the leading cause of blindness in the working age population. In the diabetic retinopathy model, the abnormal expression of P-element-induced wimpy testis-interacting RNAs (piRNAs) are involved in angiogenesis and cell proliferation pathways, suggesting that piRNAs have a certain regulatory function in diabetic retinopathy.