Huang Xia, Zeng Yi, Jiang Yujie, Qin Yueqiu, Luo Weigui, Xiang Shulin, Sooranna Suren R, Pinhu Liao
The First Clinical Medical College of Jinan University, Guangzhou, Guangdong Province 510630, China.
Department of Respiratory Medicine, Youjiang Medical University for Nationalities, Baise, Guangxi Zhuang Autonomous Region 533000, China.
Mediators Inflamm. 2017;2017:9734837. doi: 10.1155/2017/9734837. Epub 2017 May 29.
LBP and fractalkine are known to be involved in the pathogenesis of ARDS. This study investigated the relationship between LBP and fractalkine in LPS-induced A549 cells and rat lung tissue in an ARDS rat model.
A549 cells were transfected with LBP or LBP shRNA plasmid DNA or pretreated with SB203580 or SC-514 following LPS treatment. An ARDS rat model was established using LPS with or without LBPK95A, SB203580, or SC-514 treatment. RT-PCR, western blotting, ELISA, immunofluorescence, coimmunoprecipitation, and immunohistochemical staining were used to study the expression of fractalkine and LBP and p38 MAPK and p65 NF-B activities.
LPS increased LBP and reduced fractalkine. LBP overexpression further decreased LPS-induced downregulation of fractalkine and p38 MAPK and p65 NF-B activation; LBP gene silencing, SB203580, and SC-514 suppressed LPS-induced downregulation of fractalkine and p38 MAPK and p65 NF-B activation in A549 cells. LBP and fractalkine in lung tissue were increased and decreased, respectively, following LPS injection. LBPK95A, SB203580, and SC-514 ameliorated LPS-induced rat lung injury and suppressed LPS-induced downregulation of fractalkine by decreasing phospho-p38 MAPK and p65 NF-B.
The results indicate that LBP downregulates fractalkine expression in LPS-induced A549 cells and in an ARDS rat model through activation of p38 MAPK and NF-B.
已知脂多糖结合蛋白（LBP）和趋化因子参与急性呼吸窘迫综合征（ARDS）的发病机制。本研究在ARDS大鼠模型中，探讨了LPS诱导的A549细胞和大鼠肺组织中LBP与趋化因子之间的关系。
LPS处理后，用LBP或LBP shRNA质粒DNA转染A549细胞，或用SB203580或SC - 514进行预处理。使用LPS建立ARDS大鼠模型，并进行LBPK95A、SB203580或SC - 514处理。采用逆转录聚合酶链反应（RT-PCR）、蛋白质印迹法、酶联免疫吸附测定（ELISA）、免疫荧光、免疫共沉淀和免疫组织化学染色，研究趋化因子和LBP的表达以及p38丝裂原活化蛋白激酶（p38 MAPK）和p65核因子κB（p65 NF-κB）的活性。
LPS使LBP增加而趋化因子减少。LBP过表达进一步降低LPS诱导的趋化因子下调以及p38 MAPK和p65 NF-κB激活；LBP基因沉默、SB203580和SC - 514抑制A549细胞中LPS诱导的趋化因子下调以及p38 MAPK和p65 NF-κB激活。LPS注射后，肺组织中的LBP增加，趋化因子减少。LBPK95A、SB203580和SC - 514改善LPS诱导的大鼠肺损伤，并通过降低磷酸化p38 MAPK和p65 NF-κB抑制LPS诱导的趋化因子下调。
结果表明，LBP通过激活p38 MAPK和NF-κB，下调LPS诱导的A549细胞和ARDS大鼠模型中趋化因子的表达。
