Protective effects of endothelial progenitor cell microvesicles carrying miR‑98‑5p on angiotensin II‑induced rat kidney cell injury

Mai,Huade; Huang,Zhihua; Zhang,Xiaodian; Zhang,Yuanyuan; Chen,Juming; Chen,Minghui; Zhang,Yunbo; Song,Yanling; Wang,Bingshu; Lin,Yunyun; Gu,Shenhong

doi:10.3892/etm.2022.11638

Protective effects of endothelial progenitor cell microvesicles carrying miR‑98‑5p on angiotensin II‑induced rat kidney cell injury

Authors:
- Huade Mai
- Zhihua Huang
- Xiaodian Zhang
- Yuanyuan Zhang
- Juming Chen
- Minghui Chen
- Yunbo Zhang
- Yanling Song
- Bingshu Wang
- Yunyun Lin
- Shenhong Gu
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Affiliations: Department of General Practice, The First Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570102, P.R. China, Hainan Medical University, Haikou, Hainan 570000, P.R. China, Key Laboratory of Tropical Cardiovascular Diseases Research of Hainan Province, Key Laboratory of Emergency and Trauma of Ministry of Education, Research Unit of Island Emergency Medicine of Chinese Academy of Medical Sciences, Hainan Medical University, Haikou, Hainan 571199, P.R. China, Department of Cardiology, The First Affiliated Hospital of Hainan Medical University, Haikou, Hainan 570102, P.R. China
Published online on: September 29, 2022 https://doi.org/10.3892/etm.2022.11638
Article Number: 702
Copyright: © Mai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

With the increasing number of patients with hypertensive nephropathy worldwide, it has posed a major threat to health and studies on its treatment and pathogenesis are imminent. The present study investigated the mechanism through which microRNA (miR)‑98‑5p in microvesicles (MVs) secreted by endothelial progenitor cells (EPCs) is involved in the repair of angiotensin II (Ang II)‑induced injury of rat primary renal kidney cells (PRKs). After isolation of rat renal cortical sections, PRKs were isolated by density gradient centrifugation and identified by immunofluorescence staining. Transmission electron microscopy identifies successful separation of Mvs. An in vitro cell injury model was constructed using Ang II. The Gene Expression Omnibus was used to analyze the differentially expressed genes between diabetic rats and normal rats, and the Kyoto Encyclopedia of Genes and Genomes was used to analyze the signaling pathways involved in these differentially expressed genes. Reverse transcription‑quantitative PCR was used to analyze the effect of EPC‑MVs on the expression of miRNA induced by Ang II, and the levels of target genes and signaling pathway‑related proteins involved were analyzed by western blot. luciferase was used to detect the targeted binding of miR‑98‑5p to insulin‑like growth factor 1 receptor (IGF1R). Enzyme‑linked immunosorbent assay was used to analyze the effect of EPC‑MVs on Ang II‑induced oxidative stress and inflammation levels on PRKs. Cell Counting Kit‑8 was used to analyze the effect of EPC‑MVs on the cell viability of PRKs induced by Ang II. The results showed that treatment of PRKs with Ang II decreased cell viability, whereas oxidative stress and inflammation were increased. However, EPC‑MVs alleviated Ang II‑induced damage of the PRKs. During this process, the Ang‑II‑induced downregulation of miR‑98‑5p was reversed by EPC‑MVs, so miR‑98‑5p may be a key factor regulating the action of EPC‑MVs. Dual‑luciferase assay confirmed that miR‑98‑5p targets IGF1R. It was subsequently demonstrated that EPC‑MVs overexpressing miR‑98‑5p promoted phosphorylation of PI3K/Akt/endothelial nitric oxide synthase (eNOS), and inhibited the oxidative stress and inflammation in PRKs, which were reversed by the overexpression of IGF1R. In conclusion, the results of the present study demonstrated that EPC‑MVs with high expression of miR‑98‑5p can activate the PI3K/Akt/eNOS pathway by regulating IGF1R, as well as protect PRKs from Ang II‑induced oxidative stress, inflammation and inhibition of cell viability.

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