Exosomal miR-140–3p and miR-143–3p from TGF-β1-treated pancreatic stellate cells target BCL2 mRNA to increase β-cell apoptosis
Exosomal miR-140–3p and miR-143–3p from TGF-β1-treated pancreatic stellate cells target BCL2 mRNA to increase β-cell apoptosis
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DOI：
10.1016/j.mce.2022.111653
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发表时间：
2022-05
影响因子：
4.1
通讯作者：
Xiangyun Zhu;Dechen Liu;Guoqing Li;Mengmeng Zhi;Ji Sun;Liang Qi;Jingbo Li;S. Pandol;Ling Li
中科院分区：
文献类型：
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作者：
Xiangyun Zhu;Dechen Liu;Guoqing Li;Mengmeng Zhi;Ji Sun;Liang Qi;Jingbo Li;S. Pandol;Ling Li
背景：慢性胰腺炎（CP）患者通常表现为胰腺纤维化，伴有β细胞团块减少。有限的研究集中在cp相关糖尿病的发展和发病机制上。包装为外泌体的mirna是β细胞功能障碍的关键调节因子。本研究旨在确定来自活化胰腺星状细胞（PSCs）的外泌体miRNA对β细胞的影响。方法采用超离心法提取小鼠PSCs细胞系上清中的染色体，并对其进行鉴定。评估转化生长因子-β1 （TGF-β1）处理的PSCs分泌的外泌体在β细胞功能中的作用。从TGF-β1处理和未处理的PSCs （T-Exo或C-Exo）中提取外泌体制备miRNA，并通过微阵列比较miRNA表达谱。然后，miR-140-3p和miR-143-3p在MIN6细胞和胰岛中过表达或抑制，以确定其分子和功能作用。结果PSCs向培养基中分泌的细胞外囊泡主要为性体。与PBS和C-Exo孵育的MIN6细胞相比，T-Exo孵育的MIN6细胞胰岛素分泌减少，细胞活力降低。MiR-140-3p和miR-143-3p在T-Exo中显著上调。增强miR-140-3p和miR-143-3p在β-细胞中的表达可降低细胞计数和活力，提高裂解caspase-3水平。在机制上，T-Exo通过靶向受体β细胞中的b细胞淋巴瘤2基因介导miR-140-3p和miR-143-3p的细胞间转移，诱导细胞死亡。结论染色体miRNA转移作为PSCs与β-细胞之间的一种通讯方式，在治疗上有一定的应用价值。
BackgroundPeople with chronic pancreatitis (CP) normally develop a fibrotic pancreas with reduced β-cell mass. Limited studies have focused on the development and pathogenesis of CP-related diabetes. MiRNAs packaged as exosomes are the key regulators of β-cell dysfunction. This study aimed to define the effect of exosomal miRNA from activated pancreatic stellate cells (PSCs) on β-cells.MethodsExosomes in the supernatants of mouse PSCs lines were extracted via ultracentrifugation and then identified. The role of exosomes secreted by transforming growth factor-β1 (TGF-β1)-treated PSCs in β-cell function was assessed. MiRNAs were prepared from exosomes extracted from TGF-β1-treated and untreated PSCs (T-Exo or C-Exo), and the miRNA expression profiles were compared by microarray. Then, miR-140–3p and miR-143–3p were overexpressed or inhibited in MIN6 cells and islets to determine their molecular and functional effects.ResultsExosomes were the predominant extracellular vesicles secreted by PSCs into the culture medium. The MIN6 cells incubated with T-Exo had less insulin secretion and lower viability than the MIN6 cells incubated with PBS or C-Exo. MiR-140–3p and miR-143–3p were notably upregulated in T-Exo. Enhancing the expression of miR-140–3p and miR-143–3p in β-cells decreased the cell count and viability and increased the cleaved caspase-3 levels. Mechanistically, T-Exo mediated the intercellular transfer of miR-140–3p and miR-143–3p by targeting the B-cell lymphoma 2 gene in recipient β-cells to induce cell death.ConclusionsExosomal miRNA transfer as a communication mode between PSCs and β-cells, which may be explored for its therapeutic utility.