Objective: The aim of this study was to explore the effect of microRNA-141 (miR-141) on the development of diabetic nephropathy (DN) and its potential mechanism.
Methods: Real-time quantitative polymerase chain reaction (RT-qPCR) was used to detect the expression level of miR-141 in peripheral blood of DN patients. Cell apoptosis was measured by flow cytometry. The expression levels of tumor necrosis factor-α (TNF-α) and interleukin-6 (IL-6) were detected by enzyme-linked immunosorbent assay. The expression level of insulin receptor substrate 2 (IRS2) was analyzed by RT-qPCR and Western blot analysis. The targeting regulatory sites were predicted by Targetscan and luciferase assay was conducted to confirm the relationship between miR-141 and IRS2. The expression levels of protein kinase B (AKT)/adenosine monophosphate protein kinase (AMPK)-related proteins were investigated by Western blot analysis.
Results: MiR-141 was upregulated in peripheral blood of DN patients (P < 0.05). Upregulation of miR-141 significantly promoted apoptosis ( P < 0.05) and the expression of TNF-α and IL-6 ( P < 0.05). However, downregulation of miR-141 inhibited cell apoptosis ( P < 0.05) and productions of TNF-α and IL-6 ( P < 0.05). Moreover, miR-141 displayed a negatively regulatory effect on IRS2 abundance, and overexpression of IRS2 reversed the inhibitory effect of miR-141 on development of DN cells ( P < 0.05). Besides, knockdown of miR-141 significantly promoted the expressions of AKT/AMPK-related proteins ( P < 0.05), which was attenuated by inhibition of IRS2 ( P < 0.05).
Conclusion: MiR-141 promoted DN progression through regulating AKT/AMPK signaling pathway by targeting IRS2.
Keywords: diabetic nephropathy; insulin receptor substrate 2; microRNA-141; protein kinase B/adenosine monophosphate protein kinase.
© 2018 Wiley Periodicals, Inc.