|
|
|
| Regulatory Study of miR-223-3p on CaSR Expression and Calcium Oxalate Crystal Adhesion Capacity in Model of Kidney Stones |
| Abudusaimaiti AILI, GUO Mingyue, GUAN Yongzhi, et al |
| Yili Xinhua Hospital, Xinjiang Uygur Autonomous Region, Xinjiang Yili 835000, China |
|
|
|
|
Abstract Objective: To investigate the regulatory effects and mechanisms of microRNA-223-3p (miR-223-3p) on the expression of calcium-sensing receptor (CaSR) and the adhesion capacity of calcium oxalate crystals in the formation of calcium oxalate kidney stones. Methods: An in vitro model of calcium oxalate kidney stones was established using rat kidney cells,NRK-52E.The expression levels of CaSR mRNA and protein were assessed using qRT-PCR and Western blot,respectively,while miR-223-3p expression was also evaluated.The target-binding relationship between miR-223-3p and CaSR was validated using a dual-luciferase reporter assay.The effects of overexpressing and silencing miR-223-3p on CaSR expression and calcium oxalate crystal adhesion capacity were analyzed. Results: In the calcium oxalate kidney stone cell model,compared to the control group,the expression of CaSR mRNA and protein was significantly downregulated (P<0.05),while miR-223-3p expression was significantly upregulated (P<0.05).The dual-luciferase reporter assay showed that,compared to the CaSR-WT+miR-NC group,the luciferase activity in the CaSR-WT+miR-223-3p group was significantly decreased (P<0.05),while there was no significant difference in luciferase activity between the CaSR-MT+miR-223-3p and CaSR-MT+miR-NC groups (P>0.05).Under optimal transfection conditions,miR-223-3p overexpression significantly downregulated the levels of CaSR mRNA and protein (both P<0.05),whereas silencing miR-223-3p significantly upregulated their levels (both P<0.05).Compared to the model+miRNA-mimic-NC group,the calcium oxalate crystal adhesion capacity was significantly increased in the model+miR-223-3p-mimic group,while it was significantly decreased in the model+miR-223-3p-inhibitor group (both P<0.05). Conclusion: miR-223-3p targets and inhibits the expression of CaSR,thereby increasing the adhesion capacity of calcium oxalate crystals.Regulating the expression of miR-223-3p may provide new insights for the prevention and treatment of calcium oxalate kidney stones.
|
|
|
|
|
|
[1] Siener R.Nutrition and kidney stone disease[J].Nutrients,2021,13(6):1917.
[2] Miller A W,Penniston K L,Fitzpatrick K,et al.Mechanisms of the intestinal and urinary microbiome in kidney stone disease[J].Nat Rev Urol,2022,19(12):695-707.
[3] Wang G,Mi J,Bai J,et al.Non-coding RNAs in kidney stones[J].Biomolecules,2024,14(2):213.
[4] Patel Y P,Pandey S N,Patel S B,et al.Haplotype of CaSR gene is associated with risk of renal stone disease in West Indian population[J].Urolithiasis,2022,51(1):25.
[5] Damgaard M V,Treebak J T.Protocol for qPCR analysis that corrects for cDNA amplification efficiency[J].STAR Protoc,2022,3(3):101515.
[6] Liu H,Yang X,Tang K,et al.Sulforaphane elicts dual therapeutic effects on renal inflammatory injury and crystal deposition in calcium oxalate nephrocalcinosis[J].Theranostics,2020,10(16):7319-7334.
[7] Gao Y,Liu D,Zhou H,et al.Identification of biomarkers and potential therapeutic targets of kidney stone disease using bioinformatics[J]. World Urol,2024,42(1):17.
[8] Qi X,Wang R,Jin L,et al.MiR-223-3p aggravates ocular inflammation in sjogren's syndrome[J].Endocr Metab Immune Disord Drug Targets,2023,23(8):1087-1095.
[9] Qian B,Liu J,Wang J,et al.Calcium sensitive receptor and claudin-14 expression in kidney tissues of two kidney stone models[J].Arch Med Sci,2022,18(1):251-256.
[10] Massy Z A,Hénaut L,Larsson T E,et al.Calcium-sensing receptor activation in chronic kidney disease:effects beyond parathyroid hormone control[J].Semin Nephrol,2014,34(6):648-659.
[11] Han X,Liu K,Gao F,et al.Hsa-miR-223-3p participates in the process of anthracycline-induced cardiomyocyte damage by regulating NFIA gene[J].Open Med,2023,18(1):20230754. |
|
|
|
2025, Vol. 31 
