Research On The Analysis Of Active Components And Biological Activity Of Four Native Tree Species In Henan Province

Native tree species are an important part of forest resources,which have wide distribution and multiple utilization directions,and have important economic,ecological and social values.Under the influence of economic benefits,ecological environment,germplasm resources quality and other factors,the germplasm resources of native tree species have been gradually destroyed and decreased,which urgently needs protection and scientific development and utilization.Antioxidant,antibacterial and antitumor ability are the new development and utilization directions and high-value research field of native tree species.Ulmus pumila,Paulownia,Sophora japonica and Osmanthus fragrans are the four representative native tree species in Henan Province,but the understanding of their active components and biological activities is not enough.In particular,the study of antioxidant,antibacterial and antitumor activities is not comprehensive,which limits the development and utilization of high value.To explore and contrast the development potential of antioxidant,antibacterial and antitumor capabilities of these native tree species,this study collected resources,analyzed volatile and non-volatile components of different extracts,and conducted systematic comparative analysis of their antioxidant,antibacterial and antitumor activities,The biological activity of native tree species resources was evaluated by TOPSIS method to provide the theoretical basis and tree species resources for the high-value utilization of native tree species resources and the selection of excellent germplasm resources in Henan Province.The main research results are as follows:(1)Ingredient extraction and analysisThe main components of native tree resources were extracted with absolute ethanol solvent using an efficient extraction method combining ultrasonic assistance and solvent extraction.Fourier infrared spectrometer was used to analyze the changes of chemical groups before and after the extraction of native tree species resources.The position and number of absorption peaks after extraction were similar to those before extraction,indicating that native tree species resources were efficiently extracted.The positions of absorption peaks of different varieties of the same tree species were roughly the same after extraction,indicating that the species of components were consistent,which was similar to the classification results of volatile and non-volatile components.The volatile components of the extract were detected by GC-MS,mainly including alcohols,phenols,aldehydes,ketones,esters,hydrocarbons,fatty acids,acids,and terpenoids.The volatile components were mostly used as chemical raw materials,and a small part was used for medicine,food additives and flavors.Nonvolatile components were detected by high resolution LC-MS,including amino acids,glycosides,flavonoids,lignin,alkaloids,sugars,terpenes,coumarins,organic acids,etc.Nonvolatile components were mostly used in medicine,and a small part was used in chemicals,cosmetics,food additives and fragrances.(2)Antioxidant activityDPPH,ABTS,OH radical scavenging ability and Fe3+ reduction ability of the four methods were used to detect the antioxidant capacity of the extract,and the scavenging rate of 50% was used as the index to compare the antioxidant capacity between different varieties.Among the extracts of U.pumila,54-U.pumila fruits and 257-U.pumila leaves had the strongest antioxidant capacity,and the maximum OH radical scavenging capacity in257-U.pumila fruits(100.58%)was higher than that in Vc(99.97%).The maximum reducing power of 257-U.pumila leaves(1.47)exceeds Vc(1.37).Among Paulownia extracts,the flowers of diploid P.fortunei and the leaves of tetraploid P.elongata leaves have the strongest antioxidant capacity,and the maximum reducing power of diploid P.fortunei flowers(1.52)exceeds Vc(1.50);the maximum ABTS radical scavenging capacity of tetraploid P.fortunei leaves(100.09%)was higher than that of Vc(99.85%).Among the extracts of S.japonica,the antioxidant capacity of S.japonica flowers extracts was stronger than leaves.The maximum ABTS radical scavenging capacity of S.japonica flowers extracts(99.98%)was higher than that of Vc(99.85%).Among the extracts of O.fragrans,O.fragrans ‘Huangchuan Jingui’ flowers and leaves had the strongest antioxidant capacity,and the maximum ABTS free radical scavenging capacity of flowers was the same as that of Vc(99.85%).The maximum DPPH radical scavenging capacity of leaves(97.51%)was higher than that of Vc(97.12%).Comprehensively comparing the antioxidant capacity of different organs of tree species,U.pumila leaves are stronger than fruits;Paulownia flowers are stronger than leaves;S.japonica flowers are stronger than leaves;O.fragrans leaves are stronger than flowers.(3)Antibacterial activityThe antibacterial activity of extracts against Escherichia coli,Salmonella,Bacillus subtilis,Bacillus cereus and Staphylococcus aureus was detected by 96-well method,and the minimum bactericidal concentration(MBC)was used as an index to comprehensively compare and analyze the antibacterial ability.In the extract of U.pumila,the antibacterial ability of 54-U.pumila fruits and 225-U.pumila leaves was the strongest,and the MBC of54-U.pumila fruits to B.subtilis was 6.25 mg/m L.The MBC of 225-U.pumila leaves to B.subtilis,B.cereus and S.aureus was 25 mg/m L.Among Paulownia extracts,the flowers of diploid P.fortunei and the leaves of tetraploid P.elongata had the strongest antibacterial ability,and the MBC of diploid P.fortunei flowers to B.cereus and S.aureus was 6.25mg/m L.The MBC of tetraploid P.elongata leaves to E.coli,B.cereus and S.aureus was 25mg/m L.The MBC of tetraploid P.fortunei flowers on S.aureus was the lowest,which was1.0625 mg/m L.Among the extracts of S.japonica,the antibacterial ability of S.japonica flowers extracts was stronger than that of leaves.The MBC of S.japonica extracts against B.cereus,S.aureus and S.japonica leaves against B.cereus were 25 mg/m L.Among the extracts of O.fragrans,O.fragrans ‘Huangchuan Dangui’ flowers and O.fragrans‘Huangchuan Jingui’ leaves had the strongest antibacterial ability,and the MBC of O.fragrans ‘Huangchuan Dangui’ flowers on E.coli and S.aureus was 25 mg/m L.MBC of O.fragrans ‘Huangchuan Jingui’ leaves to 5 tested bacteria was 50 mg/m L.Comprehensively comparing the antibacterial ability of different organs of tree species,U.pumila fruits are stronger than leaves;Paulownia flowers are stronger than leaves;S.japonica flowers are stronger than leaves;O.fragrans leaves are stronger than flowers.(4)Antitumor activity in vitroCCK8 method was used to detect the antitumor activity of extracts on cervical cancer cells(Hela),liver cancer cells(Hep G2),breast cancer cells(MCF-7)and gastric cancer cells(SGC-7901).The cell survival rate was used as an index to comprehensively compare and analyze the antitumor ability.Among the U.pumila extracts,225-U.pumila fruits and 257-U.pumila leaves had the strongest antitumor effect,and the lethal rates of SGC-7901 cells were99.14% and 98.20%.257-U.pumila fruits had the strongest inhibitory effect on SGC-7901 cells,and the lethal rate was only 99.57%.Among Paulownia extracts,tetraploid P.fortunei flowers and diploid Paulownia-1 leaves had the strongest antitumor ability,and the lethal rates of SGC-7901 cells were 97.48% and 86.74%.The diploid P.fortunei leaves had a proliferation effect on Hela cells,and the survival rate was 122.20%.Tetraploid P.fortunei leaves had a proliferation effect on Hep G2 cells with a survival rate of 100.87%.Among the extracts of S.japonica,the antitumor ability of S.japonica leaves on each tumor cell was stronger than that of flowers.The lethal rates of S.japonica leaves on four tumor cells were98.92%,98.97%,98.45% and 99.07%.Among the extracts of O.fragrans,O.fragrans‘Huangchuan Dangui’ flowers and leaves had the strongest antitumor ability.The lethal rate of flowers to Hela cells was 97.93%,and the lethal rate of Hep G2 cells was98.15%.Comprehensively comparing the antitumor ability of different organs of tree species,U.pumila fruits are stronger than leaves;Paulownia flowers are stronger than leaves;S.japonica leaves are stronger than flowers;O.fragrans leaves are stronger than flowers.(5)Evaluation and comparative analysis of the biological activityMulti-index evaluation of antioxidant,antibacterial and antitumor biological activities of different varieties of U.pumila,Paulownia and O.Fragrans by TOPSIS method,The scores of 257-U.pumila(1.6677),tetraploid P.fortunei(1.1708)and O.fragrans‘Huangchuan Dangui’(1.1056)were the highest,which had the potential to promote breeding,processing and utilization as excellent germplasm resources.The score of S.japonica flowers(0.5858)was higher than that of S.japonica leaves,and the utilization value of S.japonica flowers was higher.A comparative analysis of single-index biological activity was carried out on the four dominant varieties.The sum of IC50 of 257-U.pumila fruits and leaves was the lowest(49.99 mg/m L),and 257-U.pumila had the strongest antioxidant activity.The sum of MBC of the flowers and leaves of the tetraploid P.elongata was the lowest(350.00 mg/m L),and the antibacterial activity of the tetraploid P.elongata was the strongest.The sum of SR of 257-U.pumila fruits and leaves was the lowest(29.26%),and 257-U.pumila had the strongest antitumor activity.The four native tree species with higher scores were evaluated by multiple indicators,and 257-U.pumila had the highest score(1.7480),and its biological activity was higher than others.Antioxidative,bacteriostatic and antitumor biological activities were correlated with volatile and non-volatile components.the results show that,the antioxidant activity had the strongest positive correlation with terpenes in non-volatile components(r=0.53),and the strongest negative correlation with esters in volatile components(r=0.36).The antibacterial activity had the strongest positive correlation with flavonoids in non-volatile components(r=0.46),and the strongest negative correlation with hydrocarbons in volatile components(r=0.49).The antitumor activity had the strongest positive correlation(r=0.42)with the terpenes in the non-volatile components,and the strongest negative correlation(r=0.45)with the carbohydrates in the non-volatile components.This paper analyzes the active ingredients of four native tree species in Henan Province,and excavates and compares the antioxidant,antibacterial and antitumor biological activities,indicating that these four local tree species have a variety of high-value utilization potential,and can be used in medicine(antioxidants,antibacterial agents,antitumor agents,etc.),chemical raw materials,food additives,etc.The multi-index evaluation of biological activity showed that the dominant varieties of four native tree species(U.pumila,tetraploid P.elongata,S.japonica,and O.fragrans ‘Huangchuan Dangui’)have potential as excellent germplasm resources.Among them,257-U.pumila can be used as key breeding and promotion resources.Components were correlated with biological activity.Non-volatile components are more correlated with biological activity,which puts forward new ideas for the purification and utilization of active components.The results of this paper provide the theoretical basis and resources for the high-value utilization of native tree species resources and the breeding and promotion of excellent germplasm resources.