Modulation of Nuclear Receptor 4A1 Expression Improves Insulin Secretion in a Mouse Model of Chronic Pancreatitis.

IF 1.7 4区医学 Q3 GASTROENTEROLOGY & HEPATOLOGY Pancreas Pub Date : 2024-10-01 Epub Date: 2024-05-04 DOI:10.1097/MPA.0000000000002370

Galande Sheethal, Archana Verma, Raghvendra Mall, Kishore Vl Parsa, Ranjeet K Tokala, Ratnakar Bynigeri, Pavan Kumar Pondugala, Krishna Vemula, S Sai Latha, Divya Tej Sowpati, Surya S Singh, G V Rao, Rupjyoti Talukdar, Thirumala-Devi Kanneganti, D Nageshwar Reddy, Mitnala Sasikala

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Abstract

Objectives: Diabetes secondary to chronic pancreatitis (CP) presents clinical challenges due to lack of understanding on factor(s) triggering insulin secretory defects. Therefore, we aimed to delineate the molecular mechanism of β-cell dysfunction in CP.

Materials and methods: Transcriptomic analysis was conducted to identify endocrine-specific receptor expression in mice and human CP on microarray. The identified receptor (NR4A1) was overexpressed in MIN6 cells using PEI linear transfection. RNA-Seq analysis of NR4A1-overexpressed (OE) MIN6 cells on NovaSeq6000 identified aberrant metabolic pathways. Upstream trigger for NR4A1OE was studied by InBio Discover and cytokine exposure, whereas downstream effect was examined by Fura2 AM-based fluorimetric and imaging studies. Mice with CP were treated with IFN-γ-neutralizing monoclonal antibodies to assess NR4A1 expression and insulin secretion.

Results: Increased expression of NR4A1 associated with decreased insulin secretion in islets (humans: controls 9 ± 0.2, CP 3.7 ± 0.2, mice: controls 8.5 ± 0.2, CP 2.1 ± 0.1 μg/L). NR4A1OE in MIN6 cells (13.2 ± 0.1) showed reduction in insulin secretion (13 ± 5 to 0.2 ± 0.1 μg/mg protein per minute, P = 0.001) and downregulation of calcium and cAMP signaling pathways. IFN-γ was identified as upstream signal for NR4A1OE in MIN6. Mice treated with IFN-γ-neutralizing antibodies showed decreased NR4A1 expression 3.4 ± 0.11-fold ( P = 0.03), showed improved insulin secretion (4.4 ± 0.2-fold, P = 0.01), and associated with increased Ca 2+ levels (2.39 ± 0.06-fold, P = 0.009).

Conclusions: Modulating NR4A1 expression can be a promising therapeutic strategy to improve insulin secretion in CP.

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调节核受体 4A1 的表达可改善慢性胰腺炎小鼠模型的胰岛素分泌。

目的：由于胰岛素分泌缺陷和相关的代谢改变，慢性胰腺炎（CP）继发的糖尿病给临床带来了挑战。由于缺乏分子方面的了解，迄今为止还没有治疗胰岛素分泌缺陷的药物疗法获得批准。我们的目的是阐明 CP 中 β 细胞功能障碍的分子机制：方法：通过芯片进行转录组分析，确定小鼠和人类 CP 中内分泌特异性受体的表达。利用 PEI 线性转染技术在 MIN6 细胞中过表达已确定的受体（NR4A1）。在 NovaSeq 6000 上对 NR4A1 过表达（OE）的 MIN6 细胞进行 RNA-Seq 分析，以确定异常代谢途径。通过 InBio Discover 和细胞因子暴露研究了 NR4A1OE 的上游触发因素。NR4A1OE 的下游效应通过基于 Fura2 AM 的荧光测定和细胞内钙成像研究进行检测。用 IFN-γ 中和单克隆抗体处理患有 CP 的小鼠，以评估 NR4A1 的表达和胰岛素分泌：结果：NR4A1表达的增加与胰岛中胰岛素分泌的减少有关（人：对照组9 ± 0.2，CP 3.7 ± 0.2；小鼠：对照组8.5 ± 0.2，CP 2.1 ± 0.1 μg/L）。MIN6 细胞（13.2 ± 0.1）中的 NR4A1OE 显示胰岛素分泌减少（13 ± 5 到 0.2 ± 0.1 μg/mg 蛋白/分钟，p = 0.001），钙和 cAMP 信号通路下调。IFN-γ 被确定为 MIN6 中 NR4A1OE 的上游信号。用 IFN-γ 中和抗体治疗的小鼠显示 NR4A1 表达下降 3.4 ± 0.11 倍（p = 0.03），胰岛素分泌改善（4.4 ± 0.2 倍，p = 0.01），同时 Ca2+ 水平升高（2.39 ± 0.06 倍，p = 0.009）：结论：调节 NR4A1 的表达是改善 CP 胰岛素分泌的一种很有前景的治疗策略。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Pancreas 医学-胃肠肝病学

CiteScore

4.70

自引率

3.40%

发文量

289

审稿时长

1 months

期刊介绍： Pancreas provides a central forum for communication of original works involving both basic and clinical research on the exocrine and endocrine pancreas and their interrelationships and consequences in disease states. This multidisciplinary, international journal covers the whole spectrum of basic sciences, etiology, prevention, pathophysiology, diagnosis, and surgical and medical management of pancreatic diseases, including cancer.