| Ganoderma lucidum is a traditional Chinese medicine-food homology fungus with a long history,which can improve health condition and prolong life.Ganoderma lucidum contains polysaccharides,amino acids,triterpenes,alkaloids and other compounds,of which polysaccharides and triterpenoids are the two major pharmacologically active compounds.In this paper,ethanol extraction and water extraction-ethanol precipitation methods were employed to isolate triterpenoids and polysaccharides.The chemical composition of triterpenoids and polysaccharides were revealed by the combination of chemical and instrumental analysis;the protective effects of triterpenoids and polysaccharides on LPS-induced acute lung injury(ALI)were compared by counting number of total cells in BALF,measuring the contents of TNF-? and IL-1? in lung tissue,serum and BALF,the content of MPO in lung tissue and analyzing the expression of inflammatory signal pathway by western blotting and EMSA.This paper aimed to further clarify the protective effects of thriterpenoids and polysaccharides on LPS-induced ALI and provide a theoretical basis for the study of Ganoderma lucidum in LPS-induced ALI.The main results obtained were as follows:(1)Triterpenoids(GT)and polysaccharides(GLP)from Ganoderma lucidum fruits,were respectively obtained by the methods of ethanol extraction,water extraction-ethanol precipitation and severg deproteinization.High performance gel permeation chromatography,liquid chromatography-mass spectrometry,infrared spectroscopy,PMP derivatization and GC-MS were used to analyze the chemical structure of active substances.GT was mainly composed of ganoderic acid C2,ganoderic acid C6,ganoderic acid G,ganoderic acid B and ganoderic acid E.GLP mainly composed of glucose and a small amount of mannose,galactose.Methylation analysis indicated that polysaccharides had different glucosidic linkage types.The main glucosidic linkage types consisted of 1-linked D-Glcp?1,3-linked D-Glcp?1,4-linked D-Glcp?1,6-linked D-Glcp?1,2,6-linked D-Glcp?1,6-linked D-Galp?1,2,6-linked D-Galp and 1,3,6-linked D-Manp(2)Comparison of the intervention effects of triterpenoids and polysaccharides on the inflammatory indexes of acute lung injury in mice: Mice were intraperitoneally administered LPS to induce acute lung injury,the weights of mice were significantly reduced,while W/D ratio and lung index,the number of total cells in the BALF and the MPO activity were significantly increased.In addition,the structure of lung tissue was damaged,showing an obvious tendency of lung injury and emphysema,a large number of inflammatory cells infiltrated lung tissue,bronchoconstriction,alveolar wall expansion of capillary hyperemia,and CD68-positive macrophages accumulated in the lung tissue.Two groups of drugs could effectively inhibit the loss of weights of mice,increase the W/D ratio and lung index,decrease the number of total cells in BALF and the content of MPO in lung tissue,alleviate the degree of lung injury,reduce inflammatory cells infiltration and accumulation of positive macrophages,and show a certain dose-dependent relationship.At the same time,the intervention effect of GLP drug group was superior to the GT drug group,which had better protective effect on acute lung injury.(3)Comparison of the intervention effects of triterpenoids and polysaccharides on the inflammatory signal pathways of acute lung injury in mice: After acute lung injury in mice,compared with the normal group,the contents of TNF-? and IL-1?,the protein expression of p-p38,p-ERK1/2,p-JNK,p-NF?Bp65 and I?B significantly increased and the DNA binding activity in NF-?B and AP-1.Compared with the model group,two drug groups could effectively decrease the contents of TNF-? and IL-1? in lung tissue,serum and BALF,inhibit MAPK and NF-?B activation,decrease the DNA-binding activity of NF-?B and AP-1,and show a dose-dependent relationship.At the same time,the intervention effect of GLP drug group was superior to the GT drug group,which has a better protective effect on acute lung injury.In conclusion,GT and GLP could alleviate the acute lung injury in mice,inhibit the expression of inflammatory cytokines,regulate the inflammatory signaling pathway and interfere with the inflammation of the lung.In addition,the intervention effect of GLP was better than that of GT. |
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