| People often rely on the forest resources for survival and hence the discovery of the potential of forest trees is an emerging trend in bio-industry.Lonicera maackii is a deciduous shrub of the genus Lonicera of Caprifoliaceae.It belongs to the same genus as honeysuckle and shows similar chemical composition and efficacy.However,compared with the widely used honeysuckle,the research and utilization of L.maackii is still limited.The research on the cytotoxicity of different parts of L.maackii from different seasons is not in depth,which seriously hinders the utilization of the forest resources.Several purpose was shown in this study,including i)evaluate and identify the active components from various parts of L.maackii,ii)identify the distribution,accumulation,types and functions of active components from different parts of L.maackii in different seasons,iii)screen and compare the in-vitro cytotoxicity of the active ingredients from different parts of L.maackii in different seasons,identify their development value as anticancer drugs,and provide useful data for people to better utilize the plant resources.The main research results are as follows:(1)The active components of volatile matter from different parts of L.maackii at different temperatures were evaluated.The volatile components of leaves,branches,barks and woods of L.maackii were analyzed by TD-GC-MS(room temperature),TGDSC(850 ℃)and PY-GC-MS(850 ℃).According to the analysis from TD-GC/MS,a total of 145 chemical components were identified from all parts of the L.maackii,and the acid compounds were the highest;The thermogravimetric results showed that the pyrolysis behaviour of different parts of L.maackii were similar.The TG analysis showed that initial and final temperature,as well as the weight loss of the four parts were different,where 20℃/min is the optimum heating rate for TG analysis;The pyrolysis results showed that 275 compounds were detected from all parts of L.maackii,and the relative content of hydrocarbons was the highest in all parts.There are differences among the active components from different parts.Valuable medical components such as falciferol,panaxynol,citronellal,etc had been identified,which confirms the presence of valuable active components in L.maackii.(2)The cytotoxicity effects of active components from L.maackii.extracts from different parts were screened.The volatile and non-volatile active components from different parts of L.maackii were detected by GC/MS and UPLC-QTOP-MS.The results showed that vitamin E,vanillic acid,carvacrol and other active components were found in the volatile components.There were 315,150,147 and 165 non-volatile chemical components detected in leaves,branches,barks and woods part.The results of colorimetric test(MTT)showed that the active component extracted using ethanol showed inhibition to different cancer cells.The analysis showed that the active component extracted from branches significantly inhibited Hepg2 cells,with the L.maackii killing rate of 85.58%;The inhibitory effect of each part on gastric cancer cells(SGC-7901)was not significant difference;The leaves had significant inhibitory effect on leukemia cells(K562),and the L.maackii killing rate was 74.08%.There were 25 types of non-volatile active ingredients was detected with medicinal effects,such as chlorogenic acid,caffeic acid,quercetin and apigenin,and its cytotoxicity were analysed.(3)The influence of seasons on the functional group of volatile chemical components from L.maackii was explored.Six parts(e.g.,leaves,branches,bark,wood,roots and root bark)from four different growth periods were selected,and the FTIR Sepctra showed that chemical component of different part from different seasons had nine common peaks but slightly different in the wavenumber and intensity.The sequence of leaf is spring>summer=autumn.The sequence of branch is the highest in winter.The sequence of wood is winter>summer>spring>autumn.The sequence of bark is spring>summer>autumn>winter.The sequence of root is the strongest in winter.The sequence of root barks is obvious in spring.The other three seasons are similar.The results showed that there were differences on the kinds or relative contents of active ingredients in different seasons,which preliminarily proved the influence of seasons on cytotoxicity.(4)The volatile active components from the extracts of L.maackii in different seasons were studied.A total of 710 chemical components was detected from all six parts of L.maackii,with spring,summer,autumn and winter detected 311,311,346 and 260 compounds,respectively.The chemical components detected from autumn L.maackii were the highest compared to other three seasons.By comparing the active components of compounds from different parts,seasons and solvents,we screened out 20 effective active components that have an impact on cytotoxicity,such as arachidonic acid of extract from leaves in autumn,3-hydroxy-4-methoxycinnamic acid from branches in spring,and explored the impact of different seasons on cytotoxicity.(5)The changes of the active components in the non-volatile components of the extracts in different seasons were studied.1870 of non-volatile components were detected from the extracts from all parts of L.maackii,and 166,180,178,and 71 active components were identified in spring,summer,autumn,and winter,respectively.Among them,there were 144,142,131 and 55 ingredients with medical use,and 21,34,29 and 14 with anti-tumor and anti-cancer properties,according to spring,summer,autumn and winter,respectively.It was found that the chemical component detected from each part was similar but the relative content was different,and different season shows differences on the active components at each parts.The characteristic of active ingredients that have an impact on cytotoxicity can be obtained accurately through principal component analysis,factor analysis and cluster analysis.(6)The effects of active components of the extracts from L.maackii on cytotoxicity were investigated.There are different inhibition effects on different cancer cells from different seasons.The inhibition effect fluctuates greatly in summer.The inhibition effect in spring,autumn and winter is strong but the difference is not obvious.Through the analysis of the distribution rule of effective active molecules and cytotoxicity results,it was determined that branches in winter had the greatest inhibitory effect on Hepg2 cells,leaves in summer on SGC-7901 cells,wood in spring on K562 cells and root bark in winter on human breast cancer cells(MDA MB-231),with the L.maackii killing rates of 79.4%,62.2%,74.4% and 70.2%,respectively.According to small molecule drug targets prediction,sesame phenol,psoralen Citric acid(also known as 3-hydroxy-4-methoxycinnamic acid)and epibetulinic acid extracted from branch in winter,β-Glutosterol,arachidonic acid,oleanolic acid and apigenin extracted from leaves in summer,psoralen isoflavones,cablin I,resinol,sesamin and citrinin extracted from wood parts in spring,aristolochic acid C,pokeweed saponin A and myristicin extracted from root bark in winter had multiple common targets on different cancer cells,which preliminarily proof the synergistic anti-cancer effect of active components from L.maackii.By evaluating and identifying the types and distribution rules of active ingredients of different parts and seasons of L.maackii,this study analyzed the effects of active ingredients of different parts and seasons on cytotoxicity,which provided valuable data for the research on anticancer effect and systematic development and utilization of L.maackii. |
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