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微小 RNA-216b 通过对 FZD5 的反向调节积极调节糖尿病血管病变。

MicroRNA-216b actively modulates diabetic angiopathy through inverse regulation on FZD5.

机构信息

Department of Cardiology, Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai 200032, China.

Department of Cardiology, Xinhua Hospital of Zhejiang Province, The Second Affiliated Hospital of Zhejiang Chinese Medical University, Hangzhou 310003, China.

出版信息

Gene. 2018 Jun 5;658:129-135. doi: 10.1016/j.gene.2018.02.050. Epub 2018 Mar 22.

DOI:10.1016/j.gene.2018.02.050
PMID:29477872
Abstract

BACKGROUND

In this work, we examined the angiogenic function of microRNA-216b in an in vitro rat diabetic model of myocardial microvascular endothelial cells (MMECs).

METHODS

MMECs were extracted from Wistar rats (MMEC(WI)) or diabetic Goto-Kakizaki (GK) rats (MMEC(GK)) and cultured in vitro. QRT-PCR was applied to compare miR-216b between MMEC(WI) and MMEC(GK). MiR-216b was downregulated in MMEC(GK). Its effects on angiogenic development, including invasion and proliferation, were evaluated. In MMEC(GK), putative miR-216b downstream target gene, frizzled class receptor 5 (FZD5), was evaluated by dual-luciferase reporter, qRT-PCR and western blot assays, respectively. FZD5 was further downregulated in MMEC(GK) with stable miR-216b downregulation to evaluate its functional role in regulating diabetic angiogenesis.

RESULTS

MiR-216b was markedly overexpressed in MMEC(GK). MiR-216b downregulation significantly enhanced angiogenesis in MMEC(GK) by promoting invasion and proliferation. FZD5 was inversely upregulated in miR-216b-downregulated MMEC(GK). Subsequently, FZD5 downregulation suppressed angiogenic development, by inhibiting invasion and proliferation in miR-216b-downregulated MMEC(GK).

CONCLUSION

MicroRNA-216b was overexposed in diabetic MMECs and its downregulation may actively enhance angiogenesis in diabetic angiopathy through inverse regulation on FZD5.

摘要

背景

在这项工作中，我们研究了 microRNA-216b 在体外大鼠糖尿病心肌微血管内皮细胞（MMECs）模型中的血管生成功能。

方法

从 Wistar 大鼠（MMEC(WI)）或糖尿病 Goto-Kakizaki（GK）大鼠（MMEC(GK)）中提取 MMECs 并进行体外培养。应用 QRT-PCR 比较 MMEC(WI)和 MMEC(GK)之间的 miR-216b。下调 MMEC(GK)中的 miR-216b。评估其对血管生成发育的影响，包括侵袭和增殖。在 MMEC(GK)中，通过双荧光素酶报告、qRT-PCR 和 Western blot 分别评估假定的 miR-216b 下游靶基因卷曲受体 5（FZD5）。在 MMEC(GK)中进一步下调 FZD5，以评估其在调节糖尿病血管生成中的功能作用。

结果

miR-216b 在 MMEC(GK)中明显过表达。下调 miR-216b 显著促进了 MMEC(GK)的血管生成，促进了侵袭和增殖。在下调 miR-216b 的 MMEC(GK)中，FZD5 呈反向上调。随后，下调 FZD5 抑制了 miR-216b 下调的 MMEC(GK)中的血管生成发育，通过抑制侵袭和增殖。

结论

糖尿病 MMECs 中 microRNA-216b 过度表达，其下调可能通过对 FZD5 的反向调节，积极增强糖尿病血管病变中的血管生成。

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