Extracellular vesicle-derived miR-146a as a novel crosstalk mechanism for high-fat induced atherosclerosis by targeting SMAD4,Journal of Advanced Research

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Extracellular vesicle-derived miR-146a as a novel crosstalk mechanism for high-fat induced atherosclerosis by targeting SMAD4
Journal of Advanced Research ( IF 11.4 ) Pub Date : 2024-08-08 , DOI: 10.1016/j.jare.2024.08.012
Kefeng Zhai ₁ , Liangle Deng ₂ , Yuxuan Wu ₂ , Han Li ₃ , Jing Zhou ₃ , Ying Shi ₃ , Jianhu Jia ₂ , Wei Wang ₄ , Sihui Nian ₅ , Ghulam Jilany Khan ₆ , Hesham R El-Seedi ₇ , Hong Duan ₃ , Lili Li ₈ , Zhaojun Wei ₉

Affiliation

School of Biological and Food Engineering, Engineering Research Center for Development and High Value Utilization of Genuine Medicinal Materials in North Anhui Province, Suzhou University, Suzhou, Anhui 234000, China; College of Biological and Food Engineering, Anhui Polytechnic University, Wuhu, Anhui 241000, China; General Clinical Research Center, Anhui Wanbei Coal-Electricity Group General Hospital, Suzhou 234000, China.
School of Biological and Food Engineering, Engineering Research Center for Development and High Value Utilization of Genuine Medicinal Materials in North Anhui Province, Suzhou University, Suzhou, Anhui 234000, China.
School of Biological and Food Engineering, Engineering Research Center for Development and High Value Utilization of Genuine Medicinal Materials in North Anhui Province, Suzhou University, Suzhou, Anhui 234000, China; College of Biological and Food Engineering, Anhui Polytechnic University, Wuhu, Anhui 241000, China.
School of Biological and Food Engineering, Engineering Research Center for Development and High Value Utilization of Genuine Medicinal Materials in North Anhui Province, Suzhou University, Suzhou, Anhui 234000, China; Department of Analytical Chemistry and Food Science, Faculty of Food Science and Technology, University of Vigo-Ourense Campus, Ourense E-32004, Spain.
School of Pharmacy, Wannan Medical College, Wuhu, Anhui 241002, China.
Department of Pharmacology and Therapeutics, Faculty of Pharmacy, University of Central Punjab, Lahore 54000, Pakistan.
International Research Center for Food Nutrition and Safety, Jiangsu University, Zhenjiang 212013, China; Department of Chemistry, Faculty of Science, Islamic University of Madinah, Madinah 42351, Saudi Arabia.
General Clinical Research Center, Anhui Wanbei Coal-Electricity Group General Hospital, Suzhou 234000, China.
School of Biological Science and Engineering, North Minzu University, Yinchuan 750021, China.

Introduction

Exosome-miR-146a is significantly increased in patients with Atherosclerosis (AS), but its mechanism and effect on AS have not been fully elucidated.

Objectives

To explore the change rule and mechanism of exosomes release, and the role and molecular mechanism of exosome-miR-146a in AS.

Methods

We isolated and identified exosomes from THP-1 macrophages after treating them with ox-LDL. Then used co-immunoprecipitation and silver staining to identify the proteins involved in regulating exosome release. PKH67 was used to label exosomes to confirm that cells can absorb them, and then co-culture with HVSMCs for cell proliferation and migration detection. The target genes of miR-146a were screened and identified through bioinformatics and luciferase activity assay, and the expression of miR-146a and related proteins was detected through qRT-PCR and Western blot in HUVECs. An AS model in LDLR^-/- mice induced by a high-fat diet was developed to investigate the impact of exosome-miR-146a on AS.

Results

The results showed that experimental foam cells from AS showed higher expression of miR-146a. It was observed that NMMHC IIA and HSP70 interacted to regulate the release of exosomes. And HUVECs can absorb exosomes derived from macrophages. In addition, we also found that miR-146a directly targeted the SMAD4 gene to modulate the p38 MAPK signaling pathway, thereby mediating HUVECs damage. Furthermore, exosome-miR-146a induced abnormal proliferation and migration of HVSMCs. The expression of miR-146a was significantly reduced in miR-146a-mimics mice and increased in miR-146a inhibitor mice whereas the inhibition of miR-146a effectively reduced while increasing miR-146a worsened AS in mice.

Conclusion

Our findings expressed the potential of miR-146a as a favorable therapeutic target for AS, however, further exploration is suggestive for deep understanding of the mechanisms regulating exosome-miR-146a release in vivo and to develop effective therapeutic strategies involving miR-146a.

中文翻译：

细胞外囊泡衍生的 miR-146a 作为靶向 SMAD4 治疗高脂肪诱导动脉粥样硬化的新型串扰机制

介绍

外泌体-miR-146a 在动脉粥样硬化（AS）患者中显著增加，但其机制和对 AS 的影响尚未完全阐明。

目标

探讨外泌体释放的变化规律和机制，以及外泌体-miR-146a 在 AS 中的作用和分子机制。

方法

我们分离并鉴定了用 ox-LDL 处理后的 THP-1 巨噬细胞的外泌体。然后使用免疫共沉淀和银染鉴定参与调节外泌体释放的蛋白质。PKH67 用于标记外泌体以确认细胞可以吸收它们，然后与 HVSMCs 共培养以进行细胞增殖和迁移检测。通过生物信息学和荧光素酶活性测定筛选鉴定 miR-146a 的靶基因，通过 qRT-PCR 和 Western blot 检测 HUVECs 中 miR-146a 和相关蛋白的表达。开发了高脂饮食诱导的 LDLR^-/- 小鼠的 AS 模型，以研究外泌体 miR-146a 对 AS 的影响。

结果

结果表明，来自 AS 的实验泡沫细胞显示 miR-146a 表达较高。据观察，NMMHC IIA 和 HSP70 相互作用调节外泌体的释放。HUVEC 可以吸收来自巨噬细胞的外泌体。此外，我们还发现 miR-146a 直接靶向 SMAD4 基因，调节 p38 MAPK 信号通路，从而介导 HUVECs 损伤。此外，外泌体 miR-146a 诱导 HVSMCs 的异常增殖和迁移。miR-146a-mimics 小鼠 miR-146a 的表达显著降低，miR-146a 抑制剂小鼠的表达升高，而 miR-146a 的抑制作用有效降低，而 miR-146a 的增加使小鼠的 AS 恶化。