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Published on: February 9, 2021
Mechanism of miRNA-141-3p in Calcium Oxalate-Induced Renal Tubular Epithelial Cell Injury via NLRP3-Mediated
Xiu-Guo Gan1, Zhi-Hao Wang1, Hai-Tao Xu1
1Department of Urology, First Affiliated Hospital of Harbin Medical University, Harbin, China.
Background/Aims:
Renal calculi represent a prevalent disorder associated with mineral deposition in renal calyces and the pelvis. Aberrant microRNA (miRNA) expression is implicated in renal injury. This study investigated the mechanism of miR-141-3p in calcium oxalate (CaOx) crystal-induced renal tubular epithelial cell (RTEC) injury.
Methods:
Human RTECs HK-2 cells were treated with CaOx crystals to induce RTEC injury. Cell viability was evaluated using Cell Counting Kit-8 assay, and apoptosis was measured using flow cytometry. The contents of lactate dehydrogenase (LDH), malondialdehyde (MDA), superoxide dismutase (SOD), interleukin (IL)-1β, and IL-18 were measured using enzyme-linked immunosorbent assay kits. The expressions of NLRP3, cleaved caspase-1, and GSDMD-N were detected using Western blot. miR-141-3p and NLRP3 expressions were determined using reverse transcription quantitative polymerase chain reaction. The binding of miR-141-3p and NLRP3 was validated using a dual-luciferase assay. The role of NLRP3 in the protection of miR-141-3p on RTEC injury was verified using functional rescue experiments.
Results:
CaOx crystals induced RTEC injury, manifested as attenuated cell viability, enhanced apoptosis, elevated intracellular LDH and MDA levels, and decreased SOD level. Pyroptosis of RTECs was enhanced by CaOx crystal induction, evidenced by elevated expressions of cleaved caspase-1, GSDMD-N, IL-1β, and IL-18. miR-141-3p expression was reduced in CaOx crystal-induced RTECs. miR-141-3p overexpression alleviated CaOx crystal-induced RTEC injury and suppressed pyroptosis of RTECs. miR-141-3p bound to NLRP3 and thereby repressed NLRP3 expression. NLRP3 overexpression reversed the protective effect of miR-141-3p overexpression on RTECs.
Conclusion:
miR-141-3p repressed NLRP3-mediated pyroptosis by suppressing NLRP3 expression, thus protecting CaOx crystal-induced RTEC injury.
Insights
MicroRNA-141-3p protects against kidney injury caused by calcium oxalate crystals by inhibiting NLRP3-mediated pyroptosis. This finding offers a potential therapeutic target for renal calculi.
Area of Science:
- Molecular Biology
- Renal Pathophysiology
- Biochemistry
Background:
- Renal calculi (kidney stones) are common and involve mineral deposition.
- Abnormal microRNA (miRNA) expression is linked to kidney damage.
- This study focuses on miR-141-3p's role in calcium oxalate (CaOx) crystal-induced renal tubular epithelial cell (RTEC) injury.
Purpose of the Study:
- To investigate the mechanism of miR-141-3p in CaOx crystal-induced RTEC injury.
- To determine if miR-141-3p can protect against RTEC damage.
- To elucidate the relationship between miR-141-3p, NLRP3, and pyroptosis in kidney stone formation.
Main Methods:
- Human RTEC (HK-2) cells were exposed to CaOx crystals.
- Cell viability, apoptosis, LDH, MDA, SOD, IL-1β, and IL-18 levels were assessed.
- Expressions of NLRP3, cleaved caspase-1, GSDMD-N, and miR-141-3p were quantified; dual-luciferase and rescue assays validated interactions.
Main Results:
- CaOx crystals impaired RTEC viability, increased apoptosis and pyroptosis markers (IL-1β, IL-18, cleaved caspase-1, GSDMD-N), and altered oxidative stress markers (LDH, MDA, SOD).
- miR-141-3p expression was downregulated in CaOx-treated RTECs.
- Overexpression of miR-141-3p mitigated CaOx-induced RTEC injury and pyroptosis by directly suppressing NLRP3 expression.
Conclusions:
- miR-141-3p acts as a protective factor against CaOx crystal-induced RTEC injury.
- It exerts its protective effect by inhibiting NLRP3-mediated pyroptosis through downregulation of NLRP3.
- Targeting miR-141-3p may offer a novel therapeutic strategy for kidney stone-related renal injury.
