Study On The Chemical Constituents And Biological Activities Of Dendrobium Huoshanense C.Z. Tang Et S.J. Cheng

Objective:The study through active screening experiments on extracts from different parts of Dendrobium huoshanense Tang et Cheng,to separate and purify the sites with better activity,in order to enrich the chemical composition of D.huoshanense Tang et Cheng,and conducted preliminary activity studies on some of the compounds isolated from them,hope to provide a theoretical basis for the development and utilization of D.huoshanense Tang et Cheng.Methods:（1）With the help of RAW 264.7 cells and 5 tumor cell models（HL-60,A-549,SMMC-7721,MCF-7,SW-480）,active screening of extracts from different parts of D.huoshanense Tang et Cheng through screening experiments of extracts of D.huoshanense Tang et Cheng on LPS-stimulated cell inflammation models（intracellular ROS level detection and activity of inhibiting NO production）and inhibition of cell proliferation activity;（2）Comprehensively utilize a variety of chromatographic separation technologies such as silica gel column chromatography,MCI column chromatography,Sephadex LH-20 column chromatography,ODS column chromatography,and high-performance liquid preparation to systematically separate and purify extracts from different parts of D.huoshanense Tang et Cheng,And use nuclear magnetic resonance(including ¹H NMR,¹³C NMR and 2D NMR),mass spectrometry,ultraviolet,infrared and other spectroscopy methods to identify the structure of the isolated compound;（3）With the help of RAW 264.7 cells and 5 tumor cell models,a part of the isolated compounds were tested for inhibitory activity of NO production through the screening experiment of inhibitory activity of NO production;With the help of 5tumor cell models,the inhibition of cell proliferation activity of some compounds was evaluated by inhibition of cell proliferation activity screening experiments.Results:（1）The experimental results show that the ethanol extract of D.huoshanense Tang et Cheng has certain pharmacological activities such as reducing the production of ROS,inhibiting the production of NO and inhibition of cell proliferation.On this basis,the ethanol extract of D.huoshanense Tang et Cheng was further processed to obtain extracts such as petroleum ether,ethyl acetate,n-butanol and water part.Perform active screening experiments on extracts from different parts,it was found that all parts of D.huoshanense Tang et Cheng had a certain effect of reducing ROS production,and the degree of inhibition increased with the increase of concentration,The inhibition effect of n-butanol part is the best;Four sites（50μg/m L）showed inhibitory activity on LPS-induced NO production in RAW 264.7 macrophages,among them,the activity of ethyl acetate is the best,and the inhibition rate of NO production is 26.39%;The petroleum ether site（200μg/m L）shows a certain biological activity against tumor cells other than HL-60 cells,the inhibition rate of A-549 cells was 49.76%,the inhibition rate of SMMC-7721 cells was 33.28%,the inhibition rate of MCF-7 cells was 31.40%,and the inhibition rate of SW-480 cells was 49.96%.（2）A total of 36 compounds were isolated from extracts from different parts of D.huoshanense Tang et Cheng,and their structures were identified as:Aduncin C（1）,5-hydroxymethylfurfural（2）,methyl 2-hydroxyglutaratey-lactone（3）,Uridine（4）,methyl paraben（5）,Michehedyoside C（6）,3,4-dihydroxy-5-methoxybenzaldehyde（7）,P-hydroxybenzoic acid（8）,5-hydroxyisobenzofuran-1（3H）-one（9）,1-methyl-5-oxo-D-proline methyl ester（10）,4-hydroxy-3 methoxy Phenylpropanol（11）,Amoenin（12）,α-dihydropicrotoxinin（13）,4-hydroxy-3,5-dimethoxybenzaldehyde（14）,3-hydroxy-4-methoxybenzaldehyde（15）,2,6-dimethoxy-p-benzoquinone（16）,dihydroferulic acid（17）,4-allyl-2,6-dimethoxyphenyl glucoside（18）,（-）-syringare sinol（19）,dendrodiumane E（20）,4,3′-dihydroxy-3,5-dimethoxybibenzyl（21）,periloyrine（22）,naringenin（23）,dihydroconiferyl dihydrop-hydroxycinnamate（24）,4,4′-dihydroxy-3,5-Dimethoxybibenzyl（25）,Aduncin（26）,12-hydroxyl-α-dihydr opicrotoxinin（27）,3,4,5-trimethoxyphenol-1-O-β-D-glucopyranoside（28）,4-O-β-D-glucopyrano-pine-aldehyde（29）,（+）-Syringaresinol（30）,（+）-syringaresinol-4-O-β-D-glucopyranoside（31）,caffeine（32）,picraquassioside C（33）,syringarein-4,4’-O-β-D-diglucoside（34）,erythrosyringoylglycerol-4-O-β-D-glucopyranoside（35）,gastrodin（36）;（3）Compound 7 and Compound 25 exhibited inhibitory activity against LPS-induced NO production in RAW 264.7 macrophages,the inhibition rate of NO production was 21.88%and 23.32%respectively;Compound 25 showed good inhibition of cell proliferation activity.the IC₅₀value of the HL-60 cells was 27.79μM,and the IC₅₀value of the SMMC-7721 cells was 6.66μM.The IC₅₀value of the of the cells was 25.01μM.Conclusion:The extracts from different parts of D.huoshanense Tang et Cheng have certain biological activities,The petroleum ether site shows good inhibition of cell proliferation activity,The ethyl acetate and n-butanol sites have a certain inhibitory effect on the production of ROS and NO in RAW 264.7 macrophages induced by LPS in mice;Separate and purify samples from different parts of D.huoshanense Tang et Cheng,a total of 36 compounds were isolated,of which compound 1 was a novel compound;The 11 compounds isolated were screened for activity.These 11 compounds all had a certain biological activity,and compound 7and compound 25 showed good inhibition of NO production,the compound 25showed good inhibition of cell proliferation activity.