Study On The Correlation Between Anti-inflammatory Components And Ecological Factors Of Iron Rod Hammer In Different Habitats In Qinghai

The plant regions affect the type and content of secondary metabolites,and the unique natural environment of Qinghai-Tibet Plateau breeds abundant medicinal plant resources.Under the background of plateau warming,investigating the effects of regions on chemical composition and biological activity of medicinal plants and their coupling relationship are helpful for comprehensive development and utilization of high-quality regional medicinal plant resources and ecological civilization construction.In our study,the wild Daodi herd Aconitum pendulum Busch from Qinghai province as the research object,the application of phytochemical and metabolomics theory survey the geographical differences of chemical composition and anti-inflammatory activity of A.pendulum Busch from different regions,and analyze the correlation among their,discuss the influence of environment parameter of the Qinghai-Tibet plateau on the chemical composition and anti-inflammatory activity of A.pendulum Busch.The main contents and results as following:1.A.pendulum Busch was rich in alkaloids and showed certain anti-inflammatory activity,and there were geographical differences in the anti-inflammatory activity of samples from difference regions.Among the 19 samples from different habitats,DTX showed the highest inhibition rate:26.00±2.50%（20μg/m L）,MYG showed the lowest inhibition rate:3.41±2.11%（20μg/m L）,which were lower than positive control:48.49±5.14%(1.00×10^-7 mol/L).The anti-inflammatory activity of A.pendulum Busch decreased with the increase of altitude,that is,the anti-inflammatory activity of A.pendulum Busch in low altitude habitat was better than that in high altitude.The anti-inflammatory activity of HZD,HZS,DTX,GLM and YSB samples was negatively correlated with sample concentration.2.Phytochemical methods were used to systematically separate and purify the alkaloid,and 38 alkaloid components were isolated and identified,including 28 C₁₉diterpenoid alkaloids,8 C₂₀ diterpenoid alkaloids and 2 other compounds,6 of which were undescribed compounds.The material basis of alkaloid component in rhirome part of A.pendulum Busch was clarified,which was used to understand the diversity of alkaloid chemical component.3.Our widely targeted metabolomics analysis identified 80 nitrogen-containing chemical compounds.Among them,pingbeimine C,neostemonine,argemonine,pallidine,norrisocorydine,armepavine,isosophocarpine,and 7,11-dehydromatrine were detected for the first time in the Aconitum genus,while szechenyianine F,pseudaconine,aldohypaconitine,flavaconitine,and 58 other alkaloids were detected in A.pendulum Busch for the first time,including diterpenoid alkaloids,apomorphline alkaloids,pyrrole alkaloids,imidazole alkaloid,steroid alkaloid,guanidine alkaloid,matrine alkaloids,promorphinane alkaloids,isoquinoline alkaloids,aromatic alkylamine alkaloids and amino acid and its derivatives,the main constituents were diterpenoid alkaloids,and the species and content of alkaloids were different in samples from different regions.Using chemometrics method,the chemical composition of A.pendulum Busch was analyzed by using volcano diagram and Venn diagram through unsupervised principal component analysis（PCA）and supervised orthogonal partial least squares discriminant analysis（OPLS-DA）.19 alkaloids were selected from 6samples of A.pendulum Busch in different habitats,which could be used as potential geographical markers.The alkaloid composition of A.pendulum Busch showed habitat difference.4.The correlation analysis of environment parameter and differential metabolic components showed that latitude and longitude were negatively correlated with Lepenine,Vilmoridine and 14-O-acetylneoline（p<0.05）,and corydine,argemonine,vilmorrianine G,etc.Elevation was positively correlated with hordenine,12-Epi-dehydronapelline,lepenine and 11-acetyllepenine,and there was a significant positive correlation with Hordenine（p<0.05）;The altitude was negatively correlated with polyschistine A,spicatine A,etc.The average temperature was positively correlated with corydine,argemonine,condelphine,etc.The mean temperature was negatively correlated with lepenine et al.The mean precipitation was negatively correlated with corydine,argemonine and 12-epi-dehydronapelline.The mean precipitation was positively correlated with hordenine,pallidine,lepenine,etc.Average sunshine time was negatively correlated with 14-O-acetylneoline and vilmoridine,and positively correlated with other differential metabolites.Orientation was negatively correlated with argemonine,condelphine,spicatine A and polyschistine A（p<0.05）,was positively correlated with other remaining differential metabolites.Correlation analysis of metabolites and anti-inflammatory activity of A.pendulum Busch from different regions showed that argemonine,polyschistine A and spicatine A were significantly positively correlated with anti-inflammatory activity of A.pendulum Busch samples.The results showed that polyschistine A and spicatine A had better anti-inflammatory activity,82.34±2.60%and 75.64±1.26%（8μM）,respectively.The correlation analysis between environment parameter and the anti-inflammatory activity of A.pendulum Busch showed that the anti-inflammatory activity of alkaloids was affected by environment parameter.The longitude and latitude,average temperature and sunshine time were positively correlated with the anti-inflammatory activity of A.pendulum Busch;Altitude,precipitation and slope were negatively correlated with the anti-inflammatory activity of A.pendulum Busch,and precipitation was the dominant factor（p<0.05）.This project is an application practice of geographical theory in the investigation of Daodi medicinal materials on the Qinghai-Tibet Plateau.Based on the regional environment,evaluating anti-inflammatory activity and phytochemical experiment for A.pendulum Busch alkaloid composition,explaining the influence of regions on chemical constituents and biological activities,the results of this study could be used to illustrate A.pendulum Busch under the influence of plateau environment and its medicinal and ecological value.It is beneficial for comprehensive utilization of high quality A.pendulum Busch resources and ecological civilization construction in Qinghai-Tibet Plateau.