Effect and mechanism of Phillyrin on acute lung injury in septic shock mice
Objective To investigate the impact of phillyrin on acute lung injury(ALI)in septic shock mice through autophagy mediated by adenosine monophosphate activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)/p70 S6 kinase(p70S6K)signal pathway.Methods Twelve mice were randomly selected as the control group,and the rest of the mice were injected intraperitoneally with 20 mg·kg-1 LPS to construct the model of septic shock,the mice with septic shock were randomly divided into a model group,an Experimental-L,-M,-H group(5 mg·kg-1,10 mg·kg-1,20 mg·kg-1 phillyrin),and an Experimental-H+compound C group(20 mg·kg-1 phillyrin+20 mg·kg-1 AMPK inhibitor compound C),there were 12 mice in each group.The lung dry weight and wet weight were weighed,and the W/D ratio was calculated;the levels of inflammatory factors tumor necrosis factor-α(TNF-α),interleukin-iβ(IL-1β),interleukin-6(IL-6)in BALF,serum endotoxin(ET)and myeloperoxidase(MPO)of lung tissue were detected by ELISA;HE staining was applied to detect pathological changes of lung tissue;Western blot was applied to detect the expression of autophagic proteins microtubule-associated protein-light chain 3(LC3-Ⅱ/Ⅰ),Beclin 1,Ras-associated GTP binding protein 7(Rab7),lysosomal associated membrane protein 2(LAMP2)and AMPK/mTOR/p70S6K signaling pathway proteins.Results In control group,model group,Experimental-L,-M,-H group and Experimental-H+compound C group LC3-Ⅱ/Ⅰ ratios were 1.43±0.14,0.73±0.07,0.81± 0.07,1.12±0.10,1.39±0.13,0.76±0.08,respectively.Beclinl protein levels were 1.05±0.11,0.43±0.05,0.50± 0.05,0.76±0.08,0.98±0.10,0.46±0.05,respectively.Rab7 protein levels were 1.53±0.17,0.67±0.06,0.70± 0.07,1.04±0.10,1.41±0.14,0.69±0.06,respectively.LAMP2 protein levels were 1.47±0.15,0.72±0.07,0.81± 0.08,1.09±0.11,1.35±0.13,0.74±0.07,respectively.p-AMPK/AMPK protein levels were 0.95±0.05,0.33± 0.03,0.39±0.04,0.68±0.07,0.91±0.09,0.36±0.04,respectively.p-mTOR/mTOR protein levels were 0.28± 0.02,0.94±0.06,0.88±0.07,0.57±0.05,0.30±0.03,0.87±0.09,respectively.The protein levels of p70S6K were 0.32±0.07,0.96±0.04,0.90±0.07,0.69±0.06,0.38±0.04,0.92±0.06,respectively.There were statistical differences between the model group and the control group(all P＜0.05).There were statistical differences between Experimental-M,-H group and model group(all P＜0.05).There were significant differences between Experimental-H+compound C group and Experimental-H group(all P＜0.05).Conclusions Phillyrin may improve ALI in septic shock mice by regulating autophagy mediated by AMPK/mTOR/p70S6K signaling pathway.
PhillyrinAMPK/mTOR/p70S6K signal pathwayAutophagySeptic shockAcute lung injury
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