Chemical Constituents Of Tripterygium Glycosides And Their Spectrum-Effect Relationship

Objective: In order to study the material basis of anti-rheumatoid arthritis activity in Tripterygium polyglycoside,a spectrum-effect relationship between the characteristic map of Tripterygium polyglycoside and anti-rheumatic arthritis was established.Methods:(1)The characteristics map of 13 batches of Tripterygium polyglycoside were established by high performance liquid chromatography(HPLC).(2)MTT assay was used to determine the toxicity of 13 batches of Tripterygium glycosides on human rheumatoid arthritis cell lines(RA-FLS)and NO release on lipopolysaccharide(LPS)-induced macrophage cell lines(RAW264.7).(3)Correlation analysis of common peaks of characteristic map and pharmacological activity was performed by partial least squares method,and the chromatographic peaks which may have biological activity were found out,and the monomer active ingredients were verified.(4)Using manual column chromatography,semi-preparative(HPLC)liquid phase,mass spectrometry,nuclear magnetic resonance and other related technologies to separate,purify and identify the main chemical constituents in Tripterygium glycosides.The MTT method is used to screen the isolated monomer compounds for anti-tumor,immunosuppression and other activities.Results:(1)The characteristic maps of 13 batches of Tripterygium glycosides were established,and a total of 30 peaks were matched.A reference map of Tripterygium glycosides was generated.The similarity between each batch and the reference map were 0.72 ? 0.96.(2)Thirteen batches of Tripterygium glycosides have different inhibitory effects on RA-FLS cells when the dose is 10 mg/L.The S3 inhibitory effect could reach 52.74%,and S13 has almost no inhibitory effect.All the13 batches of Tripterygium glycosides had different inhibitory effects on the NO release rate of RAW264.7 cells induced by LPS.The inhibitory effect of S3 was up to62.00%.(3)16 common chromatographic peaks have a positive correlation with theinhibition of RA-FLS activity;15 common chromatographic peaks have a positive correlation with the inhibition of NO release rate of RAW264.7 cells induced by LPS.Chromatographic peak 2 was identified as triptolide,chromatographic peak 10 was triptonide,chromatographic peak 21 was celastrol,and chromatographic peak 22 was wilforlide A.The contribution rate of the four compounds to the two cell models was2> 10> 21> 22.(4)Thirteen monomer compounds were isolated from Tripterygium glycosides,namely a/b-amyrenone(1),3?-acetoxyolean-12-en-28-oic acid(2),antriptolactone(3),?-hydroxypropioquaiacone(4),3-(4-hydroxy-3-methoxyphenyl)-propenal(5),3-methoxy-4-hydroxy phenylethanol(6),vanillin(7),3,4,5-trimethoxyphenol(8)p-hydroxybenzoic acid(9),p-hydroxybenzaldehyde(10),vanilyl alcohol(11),2,6-dimethxy-1,4-benzoquinone(12),bis(2-ethylhexyl)benzene-1,2-dicarboxylate(13).Compounds 1,2,5,and 12 were isolated for the first time in this genus.Compounds 1,5,and 12 at a dose of 50 ?mol/L have a significant inhibitory effect on K562 cells.Compounds 2,3,5,and 12 have a significant inhibitory effect on HEL cells.Compounds 3 and 12 have strong inhibitory effect on SH-SY5 Y cell line.Compounds 1,2,3 and 12 has obvious inhibitory effect on RA-FLS cells.Compound 12 has no cytotoxicity to HL-7702.Among them,the biological activity of compound 12 on SH-SY5 Y,HEL,RA-FLS,and HL-7702 cells was discovered for the first time.Conclusion: This paper has comprehensively studied the chemical constituents and biological activities of Tripterygium glycosides,and analyzed the correlation between their activities and characteristics map which provided experimental basis and new research strategies for the improvement of the quality control standard of Chinese medicine Tripterygium glycosides and the drug development in anti-rheumatoid arthritis.