Long non-coding RNA CDKN2B-AS1 regulates high glucose-induced human mesangial cell injury via regulating the miR-15b-5p/WNT2B axis

Abstract

Background Long non-coding RNA cyclin-dependent kinase inhibitor 2B antisense RNA 1 (CDKN2B-AS1) has been reported to be related to diabetic nephropathy (DN) progression. However, the regulatory mechanisms of CDKN2B-AS1 in DN are unclear. Methods High glucose (HG) was used to induce human mesangial cells (HMCs) for establishing the DN model. Expression levels of CDKN2B-AS1, microRNA (miR)-15b-5p, wingless-Type family member 2B (WNT2B) mRNA in serum and HMCs were detected through quantitative real-time polymerase chain reaction (qRT-PCR). The viability and cell cycle progression of HMCs were determined with Cell Counting Kit-8 (CCK-8) or flow cytometry assays. The levels of several proteins and inflammatory factors in HMCs were analyzed by western blotting or enzyme-linked immunosorbent assay (ELISA). The relationship between CDKN2B-AS1 or WNT2B and miR-15b-5p was verified with dual-luciferase reporter assay. Results CDKN2B-AS1 and WNT2B were upregulated while miR-15b-5p was downregulated in serum of DN patients and HG-treated HMCs. CDKN2B-AS1 inhibition reduced HG-induced viability, cell cycle progression, ECM accumulation, and inflammation response in HMCs. CDKN2B-AS1 regulated WNT2B expression via competitively binding to miR-15b-5p. MiR-15b-5p inhibitor reversed CDKN2B-AS1 knockdown-mediated influence on viability, cell cycle progression, ECM accumulation, and inflammation response of HG-treated HMCs. The repressive effect of miR-15b-5p mimic on viability, cell cycle progression, ECM accumulation, and inflammation response of HG-treated HMCs was abolished by WNT2B overexpression. Conclusion CDKN2B-AS1 regulated HG-induced HMC viability, cell cycle progression, ECM accumulation, and inflammation response via regulating the miR-15b-5p/WNT2B axis, provided a new mechanism for understanding the development of DN.

Long noncoding RNA X-inactive specific transcript regulates NLR family pyrin domain containing 3/caspase-1-mediated pyroptosis in diabetic nephropathy

“…Similarly, a previous study revealed that forced expression of miR-15b-5p restrained HG-triggered apoptosis of podocytes and inflammatory responses and repressed oxidative stress damage[ 43 ]. The miR-15b-5p mimic repressed the viability and inflammation of HG-treated HMCs, as previously shown[ 42 ]. The findings show that silencing XIST inhibits HG-induced RTEC pyroptosis in vitro by promoting miR-15b-5p.…”

“…qRT-PCR showed that miR-15b-5p in the DN models was significantly lower but increased after silencing XIST. Accordingly, miR-15b-5p was found to be downregulated in the serum of DN patients and in HG-treated HMCs[ 42 ]. XIST can adsorb and bind to miR-15b-5p to reduce miR-15b-5p expression in DN.…”

LncRNA HOTAIR facilitates high glucose-induced mesangial cell proliferation, fibrosis and oxidative stress in diabetic nephropathy via regulating miR-147a/WNT2B axis

“…showed that WNT2B was upregulated in DN mice and HG-induced mesangial cells, which might accelerate DN progression by enhancing cell proliferation, inflammation, and ECM accumulation [ 30 ]. Also, lncRNA CDKN2B increased WNT2B expression to promote HG-induced HMC proliferation and ECM accumulation via sponging miR-15b-5p [ 31 ]. In this, we confirmed that WNT2B could be targeted by miR-147a, and its expression was positively regulated by lncRNA HOTAIR.…”

The complex role of Wnt ligands in type 2 diabetes mellitus and related complications

“… 36 Moreover, Wnt2b is up‐regulated in serum of patients with DN and mesangial cells cultured in high glucose environment, and high glucose treatment could enhance the inflammation and extracellular matrix via activating the Wnt2b/β‐catenin pathway induced by two different long non‐coding RNAs, providing new mechanisms for understanding the development of DN and promising target for its treatment. 37 , 38 …”

“…The expression of Wnt2b and mediated canonical Wnt pathway is induced and inversely correlated with insulin expression in islets of T2DM patients, indicating inhibition of this pathway might be a new route to prevent the failure of β‐cells 36 . Moreover, Wnt2b is up‐regulated in serum of patients with DN and mesangial cells cultured in high glucose environment, and high glucose treatment could enhance the inflammation and extracellular matrix via activating the Wnt2b/β‐catenin pathway induced by two different long non‐coding RNAs, providing new mechanisms for understanding the development of DN and promising target for its treatment 37,38 …”