Emergency Department, Beijing Tongren Hospital (South District), Beijing 100176, PR China.
Emergency Department, Beijing Tongren Hospital (South District), Beijing 100176, PR China.
Immunol Lett. 2021 Mar;231:18-25. doi: 10.1016/j.imlet.2020.12.015. Epub 2021 Jan 5.
The emphasis of this study focused on the possible implication and the mechanism of miR-144-3p in septic acute lung injury (ALI) condition.
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.
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.
miR-144-3 can promote LPS induced septic ALI through downregulating Caveolin-2 to activate the JAK/STAT signal pathway.
本研究重点关注 miR-144-3p 在脓毒症急性肺损伤（ALI）中的可能作用及其机制。
在 LPS 诱导的脓毒症模型建立前，用 miR-144-3p 激动剂、miR-144-3p 拮抗剂、sh-Caveolin-2 或 PBS 预先注射小鼠。计算肺组织湿重与体重（W/W）的比值。通过 H&E 染色观察肺组织的病理变化。通过 ELISA 测定小鼠血清和肺组织中炎症细胞因子（TNF-α、IL-1β 和 IL-6）的分泌。通过 TUNEL 染色和 H&E 染色测定细胞凋亡和细胞形态。通过 qRT-PCR 和/或 Western blot 测定 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检测 miR-144-3p 与 Caveolin-2 的结合。
LPS 导致 W/W 增加、肺组织破坏、炎症反应增强和细胞凋亡。LPS 处理后，miR-144-3p 上调，Caveolin-2 下调。miR-144-3p 拮抗剂注射可减轻 LPS 诱导的炎症和细胞凋亡，但 miR-144-3p 激动剂或 sh-Caveolin-2 处理则增强了炎症和细胞凋亡。miR-144-3p 可负向靶向 Caveolin-2。miR-144-3p 可通过 Caveolin-2 激活脓毒症 ALI 小鼠中的 JAK/STAT 信号通路。
miR-144-3 通过下调 Caveolin-2 激活 JAK/STAT 信号通路，促进 LPS 诱导的脓毒症 ALI。
