PLCε1 mediates one-lung ventilation injury by regulating the p38/RhoA/NFκB activation loop.
PLCε1 mediates one-lung ventilation injury by regulating the p38/RhoA/NFκB activation loop.
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PLCγ1 通过调节 p38/RhoA/NFκB 激活环来介导单肺通气损伤。
DOI：
10.1016/j.molimm.2021.02.013
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发表时间：
2021-03
影响因子：
3.6
通讯作者：
Rui-Liu
中科院分区：
文献类型：
--
作者：
Xin-Guo;Yong-Yang;Jia-Qin Ma;Xi-Zou;Li-Sha Li;Yan-Hua Li;Yu-Zhen Hu;Rui-Liu
磷脂酶Cε -1 （PLCε1）可能是治疗炎症的一个新的潜在靶点。在本研究中，我们旨在阐明PLCε1是否参与单肺通气（OLV）所致的肺损伤，并阐明PLCε1介导的信号通路在OLV诱导的炎症反应和损伤中的潜在分子机制。方法将SD大鼠分为宽型（PLCε1-WT）组和PLCε1-KO组，分别给予OLV治疗0.5 h、1 h、2 h。采用苏木精和伊红（HE）染色和湿/干（W/D）无线电法观察大鼠肺组织损伤。此外，转染PLCε1-si RNA的肺微血管内皮细胞（PMVECs）受到脂多糖（LPS）的刺激。为了探讨PLCε1在OLV诱导的炎症损伤中的可能作用及其通路，我们制备OLV大鼠肺组织、支气管肺泡灌洗液（BALF）及pmvec进行进一步分析。采用酶联免疫分析法（ELISA）检测促炎因子的表达。反转录定量聚合酶链反应（RT-qPCR）和western blot检测相关通路蛋白NF-κB、phospho-p38、p38、phospho-ERK1/2、ERK1/2、RhoA和ROCK的活性。结果与PLCε1-WT大鼠相比，PLCε1-KOrats明显减轻了肺部炎症反应，表现为肺干湿比显著降低，促炎介质表达减少，支气管肺泡灌洗液（BALF）中性粒细胞数量和蛋白浓度下降。此外，OLV诱导的PLCε1-KO大鼠RhoA和NF-κB p65 mRNA表达升高明显受到抑制。在LPS处理的PMVECs中，PLCε1-si RNA转染的PMVECs也显示出促炎介质的表达减少，p38磷酸化水平降低，RhoA/ROCK信号激活下调。与PLCε1-si RNA和BTRB796 （p38抑制剂）共同培养在lps刺激的pmves中，RhoA和NF-κB活性显著降低。此外，ROCK抑制剂（Y-27632）或RhoA显性负突变体（RhoT19 N）均可显著降低PLCε1-si RNA处理的pmvec中NF-κB的表达。结论PLCε1在OLV诱导的炎症反应中起重要作用。此外，PLCε1通过促进p38/RhoA/ROCK激活环，促进NF-κB活化，从而增加炎症介质的表达，诱导PMVECs炎症及随后的损伤。本研究结果为降低OLV患者的炎症反应提供了一个潜在的治疗靶点。
BackgroundPhospholipase C epsilon-1 (PLCε1) might be a novel and potential target in treating inflammatory conditions. In the present study, we aimed to clarify whether PLCε1 is involved in lung injury caused by one-lung ventilation (OLV) and to elucidate the potential molecular mechanism of PLCε1-mediated signaling pathway on OLV induced inflammatory response and injury.MethodsMale Sprague-Dawley (SD) rats were divided into wide-type (PLCε1-WT) group and PLCε1-KO group, and were treated with OLV for 0.5 h, 1 h, and 2 h respectively. Observation of lung tissue injury in rats was performed by Hematoxylin and eosin (HE) staining and Wet/dry (W/D) radios. In addition, pulmonary microvascular endothelial cells (PMVECs) transfected with PLCε1-si RNA, were stimulated by lipopolysaccharide (LPS). To explore the possible roles of PLCε1 in the OLV induced inflammatory injury and the involved pathway underlying, the lung tissue and bronchoalveolar lavage fluids (BALF) of OLV rats, as well as the PMVECs were prepared for further analysis. Enzyme-linked immunoassay (ELISA) was used to detect the expression of pro-inflammatory factors. The activities of related pathway proteins (NF-κB, phospho-p38, p38, phospho-ERK1/2, ERK1/2, RhoA and ROCK) were also detected by reverse transcription quantitative polymerase chain reaction (RT-qPCR) and western blot analysis.ResultsCompared to the PLCε1-WT rats, PLCε1-KOrats exhibited marked alleviation of lung inflammation as shown by great reduction in lung wet/dry weight ratios, decreases in the expressions of pro-inflammatory mediators, and declines in the number of neutrophils and the protein concentration in bronchoalveolar lavage fluid (BALF). Moreover, the increased expressions of RhoA and NF-κB p65 mRNA induced by OLV were significantly inhibited in PLCε1-KO rats. In LPS treated PMVECs, PLCε1-si RNA transfection ones also showed the decrease expression of proinflammatory mediators, reduction in p38 phosphorylation levels and downregulation of RhoA/ROCK signaling activation. Co-cultured with PLCε1-si RNA and BTRB796 (p38 inhibitors) in LPS-stimulated PMVECs resulted in a significant reduction in RhoA and NF-κB activity. In addition, treatment with either ROCK inhibitor (Y-27632) or dominant negative mutant of RhoA (RhoT19 N) significantly reduced the expression of NF-κB in PLCε1-si RNA treated PMVECs.ConclusionThe results indicated that PLCε1 played an important role in the inflammatory response induced by OLV. Moreover, through promoting p38/RhoA/ROCK activation loop, PLCε1 promoted NF-κB activation and thereby increased the expressions of inflammatory mediators, which induced the PMVECs inflammation and subsequent injury. The results of this study provide a potential therapeutic target for the reduction of inflammatory response in patients with OLV.
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影响因子：
3.1
作者：
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