Abstract
BackgroundLncRNA NNT-AS1 (NNT-AS1) has been extensively studied as the causative agent in propagation and progression of lung and bladder cancers, and cholangiocarcinoma. However, its significance in proliferation and inflammation of diabetic nephropathy is enigmatic. This study focuses on the molecular mechanisms followed by NNT-AS1 to establish diabetic nephropathy (DN) and its potential miRNA target.MethodsBioinformatics analysis to identify potential miRNA target of NNT-AS1 and smad4 transcription factor was conducted using LncBase and TargetScan, and was subsequently confirmed by luciferase reporter assay. Relative quantitative expression of NNT-AS1 in human glomerular mesangial cells (HGMCs) was detected through quantitative real-time PCR and WB analysis. Cell proliferation was detected through CCK-8 assay, whereas, ELISA was conducted to evaluate the expression of inflammatory cytokines. Following this, relative expression of miR-214-5p and smad4 were confirmed through qRT-PCR and western blot analysis.ResultsResults from the experiments manifested up-regulated levels of NNT-AS1 and smad4 in the blood samples of DN patients as well as in HGMCs, whereas, downregulated levels of miR-214-5p were measured in the HGMCs suggesting the negative correlation between NNT-AS1 and miR-214-5p. Potential binding sites of NNT-AS1 showed miR-214-5p as its direct target and NNT-AS1 as potential absorber for this microRNA, in turn increasing the expression of transcription factor smad4.ConclusionThe data suggests that NNT-AS1 can be positively used as a potential biomarker and indicator of DN and causes extracellular matrix (ECM) accumulation and inflammation of human mesangial cells.
Highlights
LncRNA NNT-AS1 (NNT-AS1) has been extensively studied as the causative agent in propagation and progression of lung and bladder cancers, and cholangiocarcinoma
Potential binding sites of NNT-AS1 showed miR-214-5p as its direct target and NNT-AS1 as potential absorber for this microRNA, in turn increasing the expression of transcription factor smad4
The results manifested that knocking-down NNT-AS1 can significantly decrease the inflammation and progression of diabetic nephropathy (DN) and can be used as a potential therapeutic target
Summary
LncRNA NNT-AS1 (NNT-AS1) has been extensively studied as the causative agent in propagation and progression of lung and bladder cancers, and cholangiocarcinoma. Diabetes mellitus is among the most notorious group of diseases causing complications and malfunctioning of kidneys leading to chronic renal failure [1,2,3]. Renal failure is most commonly characterized by diabetic nephropathy (DN) which accumulates the extracellular matrix in tubulointerstitial and glomerular regions of the kidney and thickens the vasculature of the renal region [4]. Besides being the major reason for kidney failure, DN increases the chances of cardiovascular diseases and is said to affect one-third patients with diabetes mellitus [5]. Pathophysiological studies suggest the interference of hemodynamic and metabolic factors with normal signalling pathways and cause disease progression [6]. Glucosedependent pathways, vasoactive hormones, and their receptors directly influence renal functioning due to diabetes and gradually lead to DN [7]. The disease is usually characterized by a gradual decrease in filtration capacity of the glomerular capsule, urea with a high concentration of albumin with up to > 300 mg/day, and constantly elevated blood pressure [8]
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