Objective

The emphasis of this study focused on the possible implication and the mechanism of miR-144−3p in septic acute lung injury (ALI) condition.

Methods

Mice were pre-injected with miR-144−3p agomir, miR-144−3p antagomir, sh-Caveolin-2 or PBS before 10 mg/kg LPS induced sepsis model establishment. The ratio of wet weight of lung tissues and body weight (W/W) was calculated. The pathological changes on lung tissues were observed by H&E staining. Secretions of inflammatory cytokines (TNF-α, IL-1β and IL-6) in both mouse serum and lung tissues were determined by ELISA. Cell apoptosis and cell morphology were measured by TUNEL staining and H&E staining. The expressions of miR-144−3p, Caveolin-2, apoptotic related proteins and JAK/STAT pathway related proteins were measured by qRT-PCR or/and Western blot. Dual luciferase reporter assay was applied to detect the binding of miR-144−3p with Caveolin-2.

Results

LPS resulted in increased W/W, disrupted lung tissue, enhanced inflammatory response and cell apoptosis. miR-144−3p was upregulated while Caveolin-2 was downregulated in response to LPS treatment. Inflammation and cell apoptosis induced by LPS can be alleviated by miR-144−3p antagomir injection, but enhanced by miR-144−3p agomir or sh-Caveolin-2 treatment. miR-144−3p can negatively target Caveolin-2. miR-144−3p can activate the JAK/STAT signal pathway through Caveolin-2 in septic ALI mouse.

Conclusion

miR-144−3 can promote LPS induced septic ALI through downregulating Caveolin-2 to activate the JAK/STAT signal pathway.

中文翻译：

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MicroRNA-144-3p 通过下调 Caveolin-2 增强 LPS 诱导的小鼠脓毒性急性肺损伤

客观的

本研究的重点集中在 miR-144-3p 在脓毒性急性肺损伤 (ALI) 状况中的可能意义和机制上。

方法

在建立 10 mg/kg LPS 诱导的脓毒症模型之前，给小鼠预注射 miR-144-3p agomir、miR-144-3p antagomir、sh-Caveolin-2 或 PBS。计算肺组织湿重与体重之比（W/W）。H&E染色观察肺组织病理变化。通过ELISA测定小鼠血清和肺组织中炎性细胞因子（TNF-α、IL-1β和IL-6）的分泌。通过TUNEL染色和H&E染色测量细胞凋亡和细胞形态。采用qRT-PCR或/和Western blot检测miR-144-3p、Caveolin-2、凋亡相关蛋白和JAK/STAT通路相关蛋白的表达。应用双荧光素酶报告基因测定来检测 miR-144-3p 与 Caveolin-2 的结合。

结果

LPS 导致 W/W 增加、肺组织破坏、炎症反应增强和细胞凋亡。miR-144-3p 上调，而 Caveolin-2 响应 LPS 处理而下调。LPS 诱导的炎症和细胞凋亡可通过注射 miR-144-3p antagomir 得到缓解，但通过 miR-144-3p agomir 或 sh-Caveolin-2 处理可增强。miR-144-3p 可以负面靶向 Caveolin-2。miR-144−3p 可以通过 Caveolin-2 激活脓毒症 ALI 小鼠的 JAK/STAT 信号通路。

结论

miR-144-3 可通过下调 Caveolin-2 激活 JAK/STAT 信号通路来促进 LPS 诱导的脓毒性 ALI。