Hsa_circ_0008360 promotes high glucose-induced damage in HK-2 cells via miR-346/WNT2B axis.

IF 5.4 2区医学 Q1 Medicine

Journal of Endocrinological Investigation Pub Date : 2024-09-01 Epub Date: 2024-03-12 DOI:10.1007/s40618-024-02326-1

L Zhang, X Wang

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Abstract

Background: Diabetic nephropathy (DN) is a leading cause of end-stage renal disease worldwide. Recent researches have shown that circular RNAs (circRNAs) could affect the progress of DN, but the mechanism is still indistinct. In this work, we explored the roles of hsa_circ_0008360 in DN.

Methods: The levels of hsa_circ_0008360, microRNA-346 (miR-346) and Winglesstype family member 2B (WNT2B) were indicated by quantitative real-time polymerase chain reaction (qRT-PCR) in DN tissues and HK2 cells. Meanwhile, the protein level of WNT2B was quantified by Western blot analysis. Besides, the function of cells was examined by Cell Counting Kit-8 (CCK8) assay, flow cytometry assay, western blot, and ELISA kit. Furthermore, the interplay between miR-346 and hsa_circ_0008360 or WNT2B was detected by dual-luciferase reporter assay.

Results: The levels of hsa_circ_0008360 and WNT2B were increased, and the miR-346 level was decreased in the serum of DN patients and HG-treated HK2 cells. For functional analysis, hsa_circ_0008360 deficiency promoted cell viability, inhibits cell apoptosis, inflammatory response, and the synthesis of related fibrotic proteins in HG-treated HK2 cells. Moreover, overexpression of miR-346 induced the proliferation and inhibit apoptosis of HG-induced HK2 cells by inhibiting WNT2B expression. In mechanism, hsa_circ_0008360 acted as a miR-346 sponge to regulate the level of WNT2B.

Conclusion: Hsa_circ_0008360 can regulate miR-346/WNT2B axis in HG-induced HK2 cells, providing an underlying targeted therapy for DN patients.

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Hsa_circ_0008360通过miR-346/WNT2B轴促进高血糖诱导的HK-2细胞损伤。

背景：糖尿病肾病（DN糖尿病肾病（DN）是全球终末期肾病的主要病因。最近的研究表明，环状 RNA（circRNA）可影响 DN 的进展，但其机制尚不明确。本研究探讨了 hsa_circ_0008360 在 DN 中的作用：方法：通过实时定量聚合酶链反应（qRT-PCR）检测 DN 组织和 HK2 细胞中 hsa_circ_0008360、microRNA-346（miR-346）和 Winglesstype 家族成员 2B（WNT2B）的水平。同时，Western 印迹分析定量检测了 WNT2B 的蛋白水平。此外，还通过细胞计数试剂盒-8（CCK8）检测、流式细胞仪检测、Western印迹和ELISA试剂盒检测了细胞的功能。此外，还通过双荧光素酶报告实验检测了 miR-346 与 hsa_circ_0008360 或 WNT2B 之间的相互作用：结果：DN患者和经HG处理的HK2细胞血清中hsa_circ_0008360和WNT2B水平升高，miR-346水平降低。在功能分析方面，缺乏 hsa_circ_0008360 会促进 HG 处理的 HK2 细胞的细胞活力，抑制细胞凋亡、炎症反应和相关纤维化蛋白的合成。此外，过表达 miR-346 可通过抑制 WNT2B 的表达，诱导 HG 诱导的 HK2 细胞增殖并抑制其凋亡。在机制上，hsa_circ_0008360作为miR-346海绵调节了WNT2B的水平：结论：Hsa_circ_0008360能调节HG诱导的HK2细胞中的miR-346/WNT2B轴，为DN患者的靶向治疗提供了基础。

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来源期刊

Journal of Endocrinological Investigation ENDOCRINOLOGY & METABOLISM-

CiteScore

8.10

自引率

7.40%

发文量

242

期刊介绍： The Journal of Endocrinological Investigation is a well-established, e-only endocrine journal founded 36 years ago in 1978. It is the official journal of the Italian Society of Endocrinology (SIE), established in 1964. Other Italian societies in the endocrinology and metabolism field are affiliated to the journal: Italian Society of Andrology and Sexual Medicine, Italian Society of Obesity, Italian Society of Pediatric Endocrinology and Diabetology, Clinical Endocrinologists’ Association, Thyroid Association, Endocrine Surgical Units Association, Italian Society of Pharmacology.

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