Effects of Connexin 32-Mediated Lung Inflammation Resolution During Liver Ischemia Reperfusion.
Effects of Connexin 32-Mediated Lung Inflammation Resolution During Liver Ischemia Reperfusion.
复制标题
肝缺血再灌注期间连接蛋白 32 介导的肺部炎症消退的作用。
DOI：
10.1007/s10620-019-06020-8
复制
发表时间：
2020
影响因子：
3.1
通讯作者：
Hei Ziqing
中科院分区：
文献类型：
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作者：
Zhang Zheng;Yao Weifeng;Yuan Dongdong;Huang Fei;Liu Yue;Luo Gangjian;Hei Ziqing
背景肝缺血再灌注（HIR）可导致肺组织炎症反应和肺屏障功能障碍.在肺中广泛表达的差距连接通讯蛋白连接蛋白32（Cx 32）参与细胞间信号传导。本研究确定了通讯蛋白Cx 32是否可以影响由HIR.MethodsMice引起的肺部炎症随机分为4组（n= 8/组）：（i）Cx 32 +/+假手术组;（ii）Cx 32 +/+HIR模型组;（iii）Cx 32-/-假手术组;（iv）Cx 32-/-HIR模型组。手术后24小时，收集肺组织用于明视野显微镜、蛋白质印迹（Cx 32、JAK 2、p-JAK 2、STAT 3、p-STAT 3）和免疫荧光（ZO-1，8-OHDG）分析。检测支气管肺泡液中白细胞介素-6（IL-6）、基质金属蛋白酶-12（MMP-12）和抗胰蛋白酶（α1-AT）水平。采用PCR方法检测肺组织mmu-miR-26 a/B的表达。Cx 32缺失可明显加重HIR所致急性肺损伤的肺功能。此外，Cx 32缺失降低了ZO-1的蛋白水平（肺功能），并增加了肺中氧化应激标志物8-OHDG的水平。Cx 32 −/−HIR模型组支气管肺泡灌洗液中IL-6和MMP-12水平显著升高，JAK 2/STAT 3通路激活，α1-AT水平降低。Cx 32 −/−HIR模型组mmu-miR-26 a/B表达明显下调。Cx 32缺失可部分通过阻断mmu-miR-26 a/B的转移并导致IL-6相关的JAK 2/STAT 3通路激活而加重肝源性肺部炎症。
BackgroundHepatic ischemia reperfusion (HIR) leads to a lung inflammatory response and subsequent pulmonary barrier dysfunction. The gap junction communication protein connexin 32 (Cx32), which is widely expressed in the lungs, participates in intercellular signaling. This study determined whether the communication protein Cx32 could affect pulmonary inflammation caused by HIR.MethodsMice were randomly allocated into four groups (n= 8/group): (i)Cx32+/+sham group; (ii)Cx32+/+HIR model group; (iii)Cx32−/−sham group; and (iv)Cx32−/−HIR model group. Twenty-four hours after surgery, lung tissues were collected for bright field microscopy, western blot (Cx32, JAK2, p-JAK2, STAT3, p-STAT3), and immunofluorescence (ZO-1, 8-OHDG) analyses. The collected bronchoalveolar fluid was tested for levels of interleukin-6 (IL-6), matrix metalloproteinase 12 (MMP-12), and antitrypsin (α1-AT). Lung mmu-miR-26a/b expression was detected using a PCR assay.ResultsIncreased expression of Cx32 mRNA and protein was noted in the lungs after HIR. Cx32 deletion significantly aggravated pulmonary function from acute lung injury induced by HIR. In addition, Cx32 deletion decreased the protein level of ZO-1 (pulmonary function) and increased the level of the oxidative stress marker 8-OHDG in the lungs. Moreover, in the Cx32−/−HIR model group, the levels of IL-6 and MMP-12 in bronchoalveolar lavage fluid were significantly increased leading to activation of the JAK2/STAT3 pathway, and decreased α1-AT levels. Furthermore, we found mmu-miR-26a/b was significantly downregulated in the Cx32−/−HIR model group.ConclusionHIR leads to acute lung inflammatory injury. Cx32 deletion aggravates hepatic-derived lung inflammation, partly through blocking the transferring of mmu-miR-26a/b and leading to IL-6-related JAK2/STAT3 pathway activation.