The Preventive Effects Of Astilbin On Acute Respiratory Distress Syndrome And Its Mechanism

Objective:This research through estabilishment the model of LPS-induced ARDS mice and LPS-stimulated HUVEC injury,to study the protective effect of Astilbin on ARDS and its related mechanisms.This will provide a new theoretical basis in the prevention and treatment of ARDS.Methods:1.In vitro study.We choose HUVECs as our experiment cells.?Cells were allocated into five groups:control group,LPS group(1 ?g/ml),and LPS plus Astilbin(12.5,25,or 50?g/ml)group.Then LPS plus Astilbin(12.5,25,or 50 ?g/ml)group added with different concentrations of Astilbin.After 24 h,cells were incubated with LPS(1 ?g/ml)for 6 h.An equal volume of DMEM was added to the control group.TNF-?,IL-6,and HPA activities in the supernatant were all measured by ELISA.?Cells were allocated into three groups:control group,LPS group(1 ?g/ml),and LPS plus Astilbin(50 ?g/ml)group.Then LPS plus Astilbin(50?g/ml)group added with Astilbin(50 ?g/ml).After 24 h,cells were incubated with LPS(1 ?g/ml)for 6 h.An equal volume of DMEM was added to the control group.The production of cytoplasmic ROS in the HUVEC was monitored using the DCFH-DA and the Mitochondrial ROS detection studies were performed using MitoSOXTM Red mitochondrial superoxide indicator.The expression of Heparan sulfate(HS)were measured by immunofluorescence.2.In vivo study:All experimental animals were randomly allocated into five groups:control group;LPS group(20 mg/kg);and LPS plus Astilbin(12.5,25,or 50 mg/kg)groups.The LPS plus Astilbin groups received different concentrations of Astilbin via intraperitoneal administration.The LPS group and the LPS plus Astilbin groups were intraperitoneally injected with LPS(20 mg/kg)to establish ARDS model.After 6 h of LPS stimulation.,the lung tissue and serum samples were harvested,HE stain with lung injury score and W/D weight ratio were measured.MPO,HPA and HS were detected by ELISA.The activation state of the key regulatory proteins(P38/p-P38,JNK/p-JNK,ERK/p-ERK)in MAPKs signaling pathways was measured by Western blot.Results:Astilbin significantly decreased inflammatory cytokine release and reduced the ptoduction of ROS both in(cytoplasm and Mitochondrial)in LPS-stimulated HUVECs.Astilbin ameliorated lung histopathological alteration and W/D ratio in LPS-induced ARDS mice.According to the results of Western blot,Astilbin inhibited MAPK activation in LPS-induced ARDS mice.The result of ELISA and immunofluorescence also showed Astilbin pretreatment could protect endothelial glycocalyx integrity via reducing the HPA activity and the production of HSConclusion:Astilbin could alleviate LPS-induced ARDS damage through reducing inflammation,relieving oxidative stress,inhibiting MAPK pathway activation and protecting the endothelial glycocalyx.