| Climate change plays an important role in the growth and development of animals and plants,which have a more significant impact on plants.Most of the plants of the Amomum Roxb.are widely used in the food and pharmaceutical industries and worthy of being researched and applied.Therefore,it is necessary to study the effects of climate change on Amomum Roxb.plants.At present,the research on Amomum Roxb.mainly focuses on its chemical constituents and pharmacological activities,but there are few reports on the germplasm resources of Amomum Roxb..Therefore,in this study,the potential suitable areas of Amomum Roxb.were predicted by the Max Ent model,and the anti-inflammatory activity in vitro of Amomum Roxb.essential oil was verified by cell model in vitro.Combined with the LPS-induced acute lung injury（ALI）model,the effect of Amomum Roxb.essential oil on the protective effect of ALI and its mechanism of action,and finally through metabolomics technology to explore the biomarkers and metabolic pathways in ALI,to provide scientific ideas for the application of Amomum Roxb.plant resources in the field of respiratory diseases such as ALI.The main contents of this research are as follows:（1）By collecting the specimen information of 26 species of Amomum Roxb.in China,the geographical distribution and richness of Amomum Roxb.in China were analyzed,and it was found that Amomum Roxb.was mainly distributed in the south of the Qinling-Huaihe line in China,among which Yunnan Province was the most prominent.and Guangxi Province with the highest biological richness.The Max Ent model was used to predict the current and future potential distribution of Amomum Roxb.in China,and it was found that the area of suitable areas for A.gagnepainii and A.kravanh decreased by 39.76% and 25.87%.The most critical bioclimatic variables for the distribution of Amomum Roxb..On this basis,we selected six species（A.villosum,A.tsaoko,A.gagnepainii and A.kravanh,A.villosum Lour.var.xanthioides and A.longiligulare）that are greatly affected by climate change for further activity studies.A.villosum,is one of the four major southern medicines and has good pharmacological activity.A.villosum,A.tsaoko,and A.kravanh are homologous plants for medicine and food,and also have high economic value and pharmacological activity.（2）The essential oil components of six species of Amomum Roxb.were extracted by steam distillation,and the chemical components of Amomum Roxb.were analyzed by GC-MS.The results found that the chemical components with relatively high content were Bornyl acetate,Camphor,Cineole and 2-phenylbutanal et al;In addition,we found that（+）-δ-Cadinene is present in all six species,（-）-a-Copaene,Germacren D and L-Caryophyllene are present in five plants,and Bornyl acetate is present in and has higher content in four Amomum Roxb.,and the results of cluster analysis also show that the six species of Amomum Roxb.have a relatively close relationship.In the study of material basis,we then explored the in vitro activity of Amomum Roxb.essential oil through LPS-induced RAW264.7 cell inflammation model.The results showed that the safe concentration of A.villosum,A.tsaoko,A.gagnepainii and A.kravanh was 100μg/m L,the safe concentration of A.villosum Lour.var.xanthioides and A.longiligulare is 10μg/m L;the expression of inflammatory mediators in cells was measured by ELISA and Western Blot experiments to evaluate their anti-inflammatory and antioxidant activities;The results of immunofluorescence and Western Blot showed that A.villosum,A.tsaoko,and A.kravanh can inhibit the nuclear translocation of p65 protein in cells,and regulate the expression of p65,TLR4 and Myd88 proteins.（3）The LPS-induced mouse ALI model was established in vivo,and the pathological changes of lung tissue were evaluated by HE staining;the detection of the wet/dry weight ratio of lung tissue and the total protein concentration in BALF indicated that the species Amomum Roxb.（A.villosum,A.tsaoko,and A.kravanh）can inhibit the increase of vascular permeability in ALI mice;in addition,it was found that the essential oils can promote the activity of MDA and MPO in lung tissue significantly increase,inhibit the expression of SOD and CAT in lung tissue,BALF and serum were detected by ELISA experiments,respectively.The expression of TNF-α,IL-1β and IL-6 indicated that the inflammatory response in ALI mice was inhibited;Western Blot and immunohistochemical results showed that A.villosum,A.tsaoko,and A.kravanh essential oil could down-regulate the expression of TLR4 and Myd88 proteins expression,and down-regulated the phosphorylation levels of p65 and IκBα proteins.（4）In addition,we collected lung tissue and serum from ALI mice,respectively,and performed non-targeted metabolomic analysis on them by LC-MS,and found 8 and 15 significantly differentially metabolized in lung and serum.We enriched 23 and 15 metabolic pathways were enriched,in which we found that arachidonic acid metabolism,amido-t RNA biosynthesis,aspartate metabolism and other pathways were involved in inflammatory response and oxidative stress in ALI mice.Conclusion: In the pharmacological activity study of Amomum Roxb.plants,we found that A.kravanh,A.longiligulare,A.gagnepainii,A.tsaoko,A.villosum and A.villosum Lour.var.xanthioides had in vitro anti-inflammatory and antioxidant activities.The plant volatile oils of A.villosum,A.tsaoko,and A.kravanh exerted a protective effect on ALI through the TLR4/Myd88/NF-κB signaling pathway.In addition,climate change will have a significant impact on the growth of Amomum Roxb.,which will lead to a significant decrease in the area of their suitable areas in the future.Therefore,we should strengthen the protection of these precious and highly medicinal Amomum Roxb.plants to prevent their suitable habitats.Regional decline or even species extinction. |
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