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白细胞介素-28A/白细胞介素-10受体β轴通过调节热休克蛋白70-1的表达，经由内皮型一氧化氮合酶/蛋白激酶B信号传导以及激活蛋白-1/核因子-κB/基质金属蛋白酶-2网络促进血管生成。

IL-28A/IL-10Rβ axis promotes angiogenesis via eNOS/AKT signaling and AP-1/NF-κB/MMP-2 network by regulating HSP70-1 expression.

作者信息

Song Jun-Hui, Hwang Byungdoo, Lyea Park Sung, Kim Hoon, Jung Soontag, Choi Changsun, Myung Lee Hwan, Yun Seok-Joong, Hyun Choi Yung, Cha Eun-Jong, Patterson Cam, Kim Wun-Jae, Moon Sung-Kwon

机构信息

Department of Food and Nutrition, Chung-Ang University, Anseong 456-756, Korea.

Department of Cosmetic Science, Hoseo University, Asan-si 31499, Republic of Korea.

出版信息

J Adv Res. 2024 Aug 9. doi: 10.1016/j.jare.2024.08.013.

DOI:10.1016/j.jare.2024.08.013

PMID:39127098

Abstract

INTRODUCTION

Angiogenesis plays a significant role in the development of tumor progression and inflammatory diseases. The role of IL-28A in angiogenesis and its precise regulatory mechanisms remain rarely elucidated.

OBJECTIVES

We report the novel regulatory role of IL-28A in physiological angiogenesis. The study aimed to elucidate the regulatory mechanisms involved in IL-28A-mediated angiogenesis and identify key genes associated with IL-28A-induced angiogenic responses.

METHODS

To know the effect of IL-28A on angiogenesis, HUVECs were applied to perform proliferation, migration, invasion, tube formation, immunoblot, and EMSA. Gene expression changes in HUVECs following IL-28A treatment were analyzed by NGS. The functional role of HSP70-1 and IL-10Rβ in IL-28A-induced angiogenic responses was evaluated using PCR and siRNA knockdown. Animal studies were conducted by aortic ring ex vivo assays, Matrigel plug in vivo assays, and immunochemistry using HSP70-1 knockout and transgenic mice models. The efficacy of IL-28A in angiogenesis was confirmed in a hind-limb ischemia model.

RESULTS

Autocrine/paracrine actions in HUVECs regulated IL-28A protein expression. Exogenous IL-28A increased the proliferation of HUVECs via eNOS/AKT and ERK1/2 signaling. IL-28A treatment promoted migration, invasion, and capillary tube formation of HUVECs through induction of the AP-1/NF-κB/MMP-2 network, which was associated with eNOS/AKT and ERK1/2 signaling. The efficacy of IL-28A-induced angiogenic potential was confirmed by aortic ring and Matrigel plug assay. HSP70-1 was identified as an IL-28A-mediated angiogenic effector gene using bioinformatics. Knockdown of HSP70-1 abolished angiogenic responses and eNOS/AKT signaling in IL-28A-treated HUVECs. IL-28A-induced microvessel sprouting formation was testified in HSP70-1-deficient and HSP70-1 transgenic mice. Flow recovery in hind-limb ischemia mice was accelerated by IL-28A injection. Finally, ablation of the IL-10Rβ gene impeded the angiogenic responses and eNOS/AKT signaling stimulated by IL-28A in HUVECs.

CONCLUSION

HSP70-1 drives the progression of angiogenesis by the IL-28A/IL-10Rβ axis via eNOS/AKT signaling and the AP-1/NF-κB/MMP-2 network.

摘要

引言

血管生成在肿瘤进展和炎症性疾病的发展中起重要作用。白细胞介素-28A（IL-28A）在血管生成中的作用及其精确调控机制仍鲜有阐明。

目的

我们报告了IL-28A在生理性血管生成中的新调控作用。本研究旨在阐明IL-28A介导的血管生成所涉及的调控机制，并鉴定与IL-28A诱导的血管生成反应相关的关键基因。

方法

为了解IL-28A对血管生成的影响，应用人脐静脉内皮细胞（HUVECs）进行增殖、迁移、侵袭、管腔形成、免疫印迹和电泳迁移率变动分析（EMSA）。通过下一代测序（NGS）分析IL-28A处理后HUVECs中的基因表达变化。使用聚合酶链反应（PCR）和小干扰RNA（siRNA）敲低技术评估热休克蛋白70-1（HSP70-1）和白细胞介素-10受体β（IL-10Rβ）在IL-28A诱导的血管生成反应中的功能作用。通过离体主动脉环试验、体内基质胶栓试验以及使用HSP70-1基因敲除和转基因小鼠模型进行免疫化学研究开展动物实验。在下肢缺血模型中证实了IL-28A在血管生成中的功效。

结果

HUVECs中的自分泌/旁分泌作用调节IL-28A蛋白表达。外源性IL-28A通过内皮型一氧化氮合酶（eNOS）/蛋白激酶B（AKT）和细胞外信号调节激酶1/2（ERK1/2）信号通路增加HUVECs的增殖。IL-28A处理通过诱导激活蛋白-1（AP-1）/核因子κB（NF-κB）/基质金属蛋白酶-2（MMP-2）网络促进HUVECs的迁移、侵袭和毛细血管管腔形成，这与eNOS/AKT和ERK1/2信号通路相关。通过主动脉环和基质胶栓试验证实了IL-28A诱导的血管生成潜能的功效。使用生物信息学鉴定出HSP70-1是IL-28A介导的血管生成效应基因。敲低HSP70-1消除了IL-28A处理的HUVECs中的血管生成反应和eNOS/AKT信号通路。在HSP70-1缺陷和HSP70-1转基因小鼠中证实了IL-28A诱导的微血管芽生形成。通过注射IL-28A加速了下肢缺血小鼠的血流恢复。最后，敲除IL-10Rβ基因阻碍了IL-28A在HUVECs中刺激的血管生成反应和eNOS/AKT信号通路。