Study On Pharmacological Activity Of Hedychium Flavum Roxb.and Related Mechanism

Hedychium flavum Roxb.is a medicinal,edible and ornamental plant.Its rhizome is a Guizhou Miao medicine and is well known as Yehansu.At present,there are very few reports on the chemical composition and pharmacological activity of H.flavum.Therefore,this study provides a basis for the development and utilization of H.flavum by analyzing its chemical composition and pharmacological activity.1.The rhizome is the main medicinal part of H.flavum.Thus,we firstly study the chemical composition and pharmacological activity of the essential oil?EO?,water extract?WE?and 70%ethanol extract?EE?from H.flavum rhizome.?1?GC-MS was used to identify the chemical constituents of EO from rhizome,77chemical constituents were identified,the main chemical constituents were:?-pipene?5.4%?,?-pinene?16.8%?,1,8-cineole?5.5%?,linalool?8.5%?,?-terpineol?26.55%?,?-terpinyl acetate?2.4%?,?-curcumene?2.0%?,E-nerolidol?11.8%?,coronarin E?20.3%?.Among them,the most abundant compound is coronarin E,which was identified from H.flavum for the first time.The content of total phenol and total flavonoids in extracts was determined by the spectrophotometer.The results showed that the total phenol content of WE of H.flavum rhizome was 50.08±0.71 mg GAE/g,and the total phenol content of EE was 57.42±0.07 mg GAE/g;The total flavonoid content of WE was 12.45±1.01 mg RE/g,and the total flavonoid content of EE was 21.83±0.33 mg RE/g.UPLC-Q-Orbitriap MS was used to identify the chemical constituents of WE and EE of H.flavum rhizome.A total of 86 chemical constituents were identified,71 compounds in WE and 43 compounds in EE were identified,including 13 phenolic compounds,respectively:L-tyrosine,p-coumaric acid,cycloolivil,ferulaldehyde,1,7-bis?4-hydroxyphenyl?-3,5-heptanediol,?-?-5'-desmethylyatein,2-methoxyestradiol,3,5-di-tert-butyl-4-hydroxybenzaldehyde,carnosol,5-[?2Z,8Z?-2,8-pentadecadien-1-yl]-1,3-benzenediol,ginkgolic acid?C13:0?,2,2'-methylenebis?4-methyl-6-tert-butylphenol?,2,7-dihydroxycadalene.All 86compounds were identified from H.flavum for the first time.?2?The antioxidant,antibacterial and enzymatic activities of the rhizome of H.flavum were studied.The results showed that the extract had a strong ability to ABTS and DPPH free radical scavenging and better than the positive control.For ABTS free radical scavenging ability,water extract was 1783.16±137.57 mg AEs/g,ethanol extract was2424.29±158.31 mg AEs/g.For DPPH free radical scavenging ability,water extract was611.13±48.36 mg AEs/g;ethanol extract was 578.62±54.10 mg AEs/g.EO had a good inhibitory effect on Proteus vulgaris and Bacillus subtilis,the diameters of the inhibition zone were 24.43?5.04 mm and 18.68?0.82 mm,respectively.The minimum inhibitory concentration for Proteus vulgaris was 78.13?g/m L,and the minimum bactericidal concentration was 156.25?g/m L.It had a strong inhibitory effect on Staphylococcus aureus and Pseudomonas aeruginosa.The diameters of the inhibition zone was11.37?1.49 mm and 10.34?2.50 mm,respectively.The inhibition effect on Escherichia coli and Enterococcus faecalis is general,and the diameter of the inhibition zone was<10.00 mm.WE and EE of H.flavum rhizome had a very strong inhibitory effect on?-glucosidase,and the inhibitory effect was better than control acarbose,the inhibitory ability of WE was 6.02±0.79 mmo L ACEs/g,the inhibitory ability of EE was 2.04±0.20mmo L ACEs/g,WE and EEt had good inhibitory effect on tyrosinase,and the inhibitory ability were 19.30±2.72 mg KAEs/g and 20.85±1.12 mg KAEs/g,respectively.?3?The cytotoxicity of EO of H.flavum rhizome was measured by MTT method.The results showed that EO had selective cytotoxicity,with significantly higher cytotoxicity to four tumor cells than to normal cell MRC-5?P<0.05?.EO had the best inhibitory effect on K562 cells.The IC₅₀value was 27.1±2.18?g/m L.After 48 hours,the cell morphological changes,AO/EB and Hoechst 33258 fluorescence staining results showed that K562 cells apoptosis.Western Blotting was used to detect the expression of related proteins after the EO acted on the cells.The results showed that the expression of Caspase-3 in cells changed,but the ratio of Caspase-3 to precursor caspase-3 did not change significantly,the expression of Caspase-8 was up-regulated?P*<0.05?,and the ratio of Caspase-8 to precursor caspase-8 was up-regulated?P*<0.05?;the ratio of Bax protein expression to Bcl-2 protein expression was up-regulated?P*<0.05?,Cyt c protein expression was significantly up-regulated?P**<0.01?,neither p53 nor MDM2 protein expression occurred significant changed.The results showed that the EO could induce apoptosis through endogenous and exogenous pathways,and activate the Caspase cascade reaction of K562 cells,thereby induced apoptosis.2.A comparative study on the chemical components and pharmacological activities of different parts of the H.flavum?rhizome,stem,flower and leaf?.The total phenol and total flavonoids and antioxidant and enzyme inhibitory activities in each part of the H.flavum were study,the results showed that the highest content of total phenol and total flavonoids were the 70%ethanol extract of leaf,and the total phenol content was 72.08±1.02 mg GAE/g,the total flavonoid content was88.16±0.80 mg RE/g.The strongest scavenging ability on ABTS and DPPH free radicals was the ethanol extract of leaf,ABTS free radical scavenging ability was 872.18±39.78BHT mg/g,DPPH radical scaveng ability was 11407.74±751.34 BHT mg/g.The strongest inhibitory effect on the selected four enzymes was the leaf ethanol extract.The best inhibitory effect on?-glucosidase was 17.32±1.18 mmo L ACEs/g.To sum up,the EO has strong antibacterial activity and selective cytotoxicity of tumor cells.The water extract and 70%alcohol extract has strong antioxidant capacity,high-efficiency?-glucosidase inhibitory effect and low cytotoxicity.The results show that H.flavum has great potential for use in cosmetics,food and medical treatment.