微小 RNA-21 通过靶向 MAP2K3/P38 MAPK 信号通路对柯萨奇病毒 B3 感染的保护作用。

The protective role of microRNA-21 against coxsackievirus B3 infection through targeting the MAP2K3/P38 MAPK signaling pathway.

机构信息

Department of Biochemistry & Immunology, Capital Institute of Pediatrics-Peking University Teaching Hospital, YaBao Road 2, Beijing, 100020, China.

Department of Biochemistry & Immunology, Capital Institute of Pediatrics, YaBao Road 2, Beijing, 100020, China.

出版信息

J Transl Med. 2019 Oct 4;17(1):335. doi: 10.1186/s12967-019-2077-y.

DOI:10.1186/s12967-019-2077-y

PMID:31585536

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC6778380/

Abstract

BACKGROUND

The P38 mitogen-activated protein kinase (MAPK) pathway plays an essential role in CVB3-induced diseases. We previously demonstrated microRNA-21 has potential inhibitory effect on the MAP2K3 which locates upstream of P38 MAPK and was upregulated in mouse hearts upon CVB3 infection. However, the effect and underlying mechanism of miRNA-21 on CVB3 infection remain unclear.

METHODS

We detected continuous changes of cellular miRNA-21 and P38 MAPK proteins expression profiling post CVB3 infection in vitro within 12 h. P38 MAPK signaling was inhibited by the specific inhibitor, small interfering RNA and miRNA-21 mimic in vitro, CVB3 replication, cell apoptosis rate and proliferation were detected. Viral load in the mice heart, cardiomyocyte apoptosis rate and histological of the heart were also detected in the mice model of viral myocarditis pretreated with miRNA-21-lentivirus.

RESULTS

We observed significant upregulation of miRNA-21 expression followed by suppression of the MAP2K3/P38 MAPK signaling in CVB3-infected Hela cells. The inactivation of the MAP2K3/P38 MAPK signaling by P38 MAPK specific inhibitor, small interfering RNA against MAP2K3, or miRNA-21 overexpression significantly inhibited viral progeny release from CVB3-infected cells. Mechanistically, when compared with control miRNA, miRNA-21 showed no effect on capsid protein VP1 expression and viral load within host cells, while significantly reversing CVB3-induced caspase-3 activation and cell apoptosis rate, further promoting proliferation of infected cells, which indicates the inhibitory effect of miRNA-21 on CVB3 progeny release. In the in vivo study, when compared with control miRNA, miRNA-21 pretreatment remarkably inactivated the MAP2K3/P38 MAPK signaling in mice and protected them against CVB3 infection as evidenced by significantly alleviated cell apoptosis rate, reduced viral titers, necrosis in the heart as well as by remarkably prolonged survival time.

CONCLUSIONS

miRNA-21 were reverse correlated with P38 MAPK activation post CVB3 infection, miRNA-21 overexpression significantly inhibited viral progeny release and decreased myocytes apoptosis rate in vitro and in vivo, suggesting that miRNA-21 may serve as a potential therapeutic agent against CVB3 infection through targeting the MAP2K3/P38 MAPK signaling.

摘要

背景

P38 丝裂原活化蛋白激酶（MAPK）通路在柯萨奇病毒 B3（CVB3）诱导的疾病中发挥重要作用。我们之前的研究表明，microRNA-21 对 MAP2K3 具有潜在的抑制作用，MAP2K3 位于 P38 MAPK 的上游，在 CVB3 感染小鼠心脏时上调。然而，miRNA-21 对 CVB3 感染的作用及其潜在机制尚不清楚。

方法

我们在体外检测了 CVB3 感染后 12 小时内细胞中 miRNA-21 和 P38 MAPK 蛋白表达谱的连续变化。通过特异性抑制剂、小干扰 RNA 和 miRNA-21 模拟物抑制 P38 MAPK 信号通路，检测 CVB3 复制、细胞凋亡率和增殖情况。在 miRNA-21-慢病毒预处理的病毒性心肌炎小鼠模型中，还检测了小鼠心脏中的病毒载量、心肌细胞凋亡率和心脏组织学变化。

结果

我们观察到在 CVB3 感染的 Hela 细胞中，miRNA-21 的表达显著上调，随后 MAP2K3/P38 MAPK 信号通路受到抑制。通过 P38 MAPK 特异性抑制剂、针对 MAP2K3 的小干扰 RNA 或 miRNA-21 过表达使 MAP2K3/P38 MAPK 信号失活，可显著抑制 CVB3 感染细胞中的病毒子代释放。从机制上讲，与对照 miRNA 相比，miRNA-21 对宿主细胞内衣壳蛋白 VP1 的表达和病毒载量没有影响，但显著逆转了 CVB3 诱导的 caspase-3 活化和细胞凋亡率，进一步促进了感染细胞的增殖，表明 miRNA-21 对 CVB3 子代释放具有抑制作用。在体内研究中，与对照 miRNA 相比，miRNA-21 预处理可显著使 MAP2K3/P38 MAPK 信号失活，并保护小鼠免受 CVB3 感染，表现为细胞凋亡率显著降低、病毒滴度降低、心脏坏死以及存活时间显著延长。