Long-coding RNA ANRIL knockdown improves diabetes-induced renal injury
Nan Su 2
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General Medicine Department, The First Affiliated Hospital of Soochow University, Suzhou, China
Department of Respiratory and Critical Care Medicine, The First Affiliated Hospital of Soochow University, Suzhou, China
Yan Xie   

The First Affiliated Hospital of Soochow University, China
Submission date: 2021-01-12
Final revision date: 2021-03-10
Acceptance date: 2021-03-11
Online publication date: 2021-03-18
The aim of the study was to investigate the effects and mechanisms of long non-coding RNA (lncRNA) in diabetes-induced renal injury, both in vivo and in vitro.

Material and methods:
Sixty healthy subjects and 30 patients with diabetic nephropathy (DN) were enrolled in this study. The lncRNA ANRIL expression was measured by RT-qPCR assay in serum. Further, the ANRIL expression in kidney tissue of normal, 10-day DN, and 20-day DN rat models was measured. ELISA was used to measure levels of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), interleukin-6 (IL-6), and tumor necrosis factor (TNF)-α. Renal pathology was evaluated by hematoxylin-eosin (HE) staining, cell apoptosis by TUNEL assay, relative protein expression by immunohistochemical (IHC) assay, and relative mRNA expression by RT-qPCR assay. In the cell experiments, LDH, MDA, SOD, IL-6, and TNF-α levels were measured by enzyme-linked immunosorbent assay (ELISA); cell proliferation by Cell Counting Kit-8 (CCK-8); cell apoptosis by flow cytometry; and relative mRNA and protein expression by real-time quantitative polymerase chain reaction (RT-qPCR) and Western blot (WB) assay. The nuclear factor κB (NF-κB) (p65) nuclear volume of difference groups was evaluated by cell immunofluorescence.

lncRNA ANRIL was significantly higher in DN patients than healthy subjects. Compared with the sham group, the ANRIL expression and LDH, MDA, IL-6 and TNF-α concentrations were significantly increased, and SOD concentrations and cell apoptosis rate were significantly decreased with increasing time. Toll-like receptor 4 (TLR4), myeloid differentiation factor 88 (MyD88), and NF-κB(p65) mRNA and protein expression levels were significantly up-regulated.

lncRNA ANRIL knockdown could improve diabetes-induced renal injury via regulation of TLR4/NF-κB(p65).