Studies On Anti-inflammatory Activity Constituents Of Sabia Parviflora And Metabolites Of H4

Sabia parviflora Wall.ex Roxb.is the stems and leaves of Sabia parviflora Wall.ex Roxb.（S.P.）belonging to the Sabia family,which is widely used in Buyi and Miao people.Sabia parviflora commonly known as small yellow drug,ya xi qiang,yellow swollen drugs and so on.It is regionally distributes in the provinces of Guangxi,Yunnan and Guizhou in China and mainly grows at an altitude of 600m to 2900m.It grows mainly in shrubs of the mountains and sparse forests beside streams.It could clear heat and drain dampness and sooth liver-gallbladder,mainly used for the treatment of hot and humid jaundice,hepatitis,bleeding and so on.It is mainly used various liver diseases,especially for the icteric hepatitis.According to previous reports,the main chemical constituents of Sabia parviflora Wall.ex Roxb are triterpenes,alkaloids,organic acid and flavonoids.These compounds determine it has many pharmacological effects,including anti-inflammation.Our research group has studied the chemical constituents of the 50%and 70%ethanol extracts of the Sabia parviflora Wall.ex Roxb.In order to further discover its active substances,30%ethanol extract was separated by silica gel column chromatography,gel column chromatography,medium pressure MCI column chromatography,semi-preparative liquid phase.and preparative liquid phase.and a total of 26 compounds were separated.Their structures were identified by ESI-MS,¹H-NMR and ¹³C-NMR techniques.The names of these compounds are as follows:3,4,5-trimethoxy benzoic acid（1）,vanillic acid（2）,Syringic acid（3）,1,3,5-trimethoxybenzene（4）,P-hydroxybenzoic acid（5）,pyrocatechol（6）,1,2,4-trihydroxybenzene（7）,（7α,8S）-erythro-1-C-syringylglycerol-4-O-β-D-glucopyranoside（8）,Ferulic acid（9）,syringicacid-4-O-β-D-glucopyranoside（10）,Syringic acid-4-O-β-D-glucopyranoside（11）,lyoniresinol（12）,rel-5-（3S,8S-dihydroxy-1R,5S-dimethyl-7-oxa-6-oxobicyclo[3,2,1]-oct-8-yl）-3-methyl-2Z,4E-penta-dienoic acid（13）,Protocatechuic acid（14）,4-quinolinone-2-caboxylic acid（15）,2,6-dimethoxy-4-hydroxybenzyl alcohol 1-O-β-D-glucopyranoside（16）,Threo-guaiacylglycerol-β-O-4’–coniferyl alcohol（17）,protocatechuic acid-3-O-b-Dxylopyranoside（18）,Rutin（19）,n-hexacos-17-enoic acid（20）,1-[1-Hydroxy-2-（dimethylamino）ethyl]-3,4-phenanthrenediol-4-O-β-D-glucose（21）,1-Deoxy-1-（3,4-dihydro-7-methyl-2,3-dioxo-1（2H）-quinoxalinyl）pentitol-6-carboxyl（22）,[7-Hydroxy-1-（4-hydroxy-3,5-dimethoxyphenyl）-3-（hydroxymethyl）-4-（hydroxy）-6,8-dimethoxy-1,2,3,4-tetrahydro-2-naphthalenyl]methyl-O-β-D-glucopyranoside（23）,4-（3-Carboxy-4-hydroxyphenyl）-2-Methyl-2-butenoic acid（24）,5-（3-Carboxy-4-hydroxyphenyl）-5-oxo-2-Methyl-2-pentenoic acid（25）,3.3-2methoxy-4-hydroxybenzoic-4-O-β-D-apiofuranosyl（1→2）-4-O-β-D-rhamnoside（26）.Among them,compounds（21-26）are new compounds,compounds（1,3-4,6-18,20）were reported from the genus Sabia for the first time.On the basis of previous work,we screened the isolated compounds according to the known activity of the Sabia parviflora Wall.ex Roxb and the existing experimental conditions.We used LPS-induced mouse macrophage（RAW264.7）to make inflammation model.We used Griess reagent to determine the content of NO produced by the compound to determine the anti-inflammatory activity of the compounds.The results showed that compound 3,4,5,8,9,17 had obvious anti-inflammatory activity,while compound 10,12,14,15,22,25 had weaker anti-inflammatory activity.Finally,as a natural alkaloid compound,H4 is one of the signature chemical components of the Sabia parviflora.Experiments have found that H4 has significant Hepatoprotective Effects and certain research significance for new drug development,but at present there is no research on H4 metabolites at home and abroad.In this experiment,Ultra-High Performance Liquid Chromatography-Quadrupole-time-of-flight Mass Spectrometry（UHPLC-QTOF-MS/MS）was used to identify H4 metabolites.We studied the metabolites of H4 in the urine,plasma,feces,and bile of rats.A total of 31 metabolites were identified in plasma,urine,bile and feces.It was found that H4 mainly undergoes hydroxylation,methylation,sulfonation,and glucuronidation in rats.Metabolites are mainly excreted from the urine.In addition this experiment provides some scientific basis for the subsequent in vivo metabolism of H4.