Effect And Application Of Hemolytic Phosphatidic Acid In The Treatment Of Acute Respiratory Distress

OBJECTIVE:To analyze the LPA receptor type of LPA in lung tissue and AECII cells in cell and animal model,screening LPA（different molecular type）activation of AECII cell LPA receptor Wave conditions;and promote the AECII survival and pharmacological mechanism by studying the LPA,and improve the therapeutic effect of LPA on ARDS.The longitudinal breakthrough in the three levels of tissue,cells and animals,is intended to clarify the molecular mechanism of LPA to promote lung-proof repair,and further clearly define ARDS treatment strategies to repair lung injury as the core of atomizing LPA.The mortality rate of Ali/ARDS provides new treatment ideas.Methods:1,clinical study:8 cases of ARDS group and health control group were taken from peripheral blood to detect inflammatory factors IL-1β,IL-6,IL-10,TNF-αcontent.2,animal research:（1）12 rats were randomly divided into 3 groups:1C group:tail vein injection with lipopolysaccharide（LPS）isometric physiological saline;2LPS group:tail vein injection 5 mg/kg of lipopolysaccharide（LPS）Applying for simulation of ARDS;3Lps+LPA group:After tail vein injection polysaccharide（LPS）,the LPA10μm atomized inhalation was carried out.^[1]Take the peripheral blood serum of each group,test inflammation IL-1β,IL-6,IL-10,TNF-αcontent.^[2]Observation of cell ultrastructure under various groups of pulmonary tissues and electron microscopy.^[2]Rat alveolar II epithelial cells were isolated and purified for subsequent cell experiments.3,Cell Studies:（1）Extraction of RT-PCR in Rt-PCR to detect LPAR1-6 expression.^[1]divide the cells into 1 normal AECII group（group C）;2LPA stimulates the AECII cell group（LPA group）;3LPAR1 inhibitor ONO stimulates the AECII cell group（ONO group）;4LPA and ONO Commonly stimulate AECII cell group（L+O group）;5WNT signaling pathway inhibitor XAV stimulates the AECII cell group（XAV group）;6LPA combined with XAV to stimulate the AECII cell group（L+X group）;RT-PCR method detected GSK-3β,beta-catenin expression,Western Blot method detectsβ-catenin expression.^[2]Stimulate AECII cells with different concentrations of LPA and detect the proliferation and migration of cells.（4）divide the cells into 1C group;2LPA group;3ono group;4LPA+ONO group;5XAV group;6LPA+XAV group;7EGF stimulate AECII cell group（EGF group）;8EGF and ONO Commonly stimulate AECII cell group（EGF+ONO group）;（9）EGF co-stimulates the AECII cell group（EGF+XAV group）;detects the proliferation and migration of various groups of cells.（5）The cells were divided into1C group;2Lps group;3LPA group;4Lps+LPA group,monitoring the inflammatory factor IL-1β,IL-6,IL-10,TNF-αcontent in each group of cell supernatants.RESULTS:Clinical study:The content of serum in serum in serum in ARDS was significantly higher than that of the health control group.Animal study:The inflammatory factor content in rat cells under LPA+LPS treatment was significantly lower than the LPS treatment group.LPA+LPS treatment under ultrastructural LPA+LPS has a certain increase in the number of micro-velvet hair,and the cell double film structure is normal,and the small body is constantly returned to normal.Cell studies:1LPAR1 significantly high expression,followed by LPAR6;LPAR3 and LPAR5 small expression,LPAR2 and LPAR4 are almost non-expressed.2 After the addition of the LPA receptor,GSK-3βandβ-catenin expression were significantly increased relative to the control group.After the addition of the LPA receptor 1 inhibitor,GSK-3βandβ-catenin expression were significantly reduced,and in LPA treatment After the addition of XAV-939 inhibitors,GSK-3βand beta-catenin have a significant decrease.The 3LPA has gradient promoting AECII cell proliferation,and this proliferation will be significantly reduced after adding Wnt signaling pathway inhibitors.4LPA has gradient promoting AECII cell proliferation,and this proliferation will be significantly reduced after adding Wnt signaling pathway inhibitors.Conclusion:1.The main expression receptor of LPA in AECII cells is LPAR1,and LPA acts on AECII cells via Wnt cellular pathway.2.LPA promotes the proliferation and migration of AECII and is mainly mediated byβ-catenin protein.3.ARDS can cause increase in anti-inflammatory factors and anti-inflammatory factors,while LPA can alleviate inflammatory factors under inflammatory conditions.