Chemical Constituents And Anti-Breast Cancer Activity Of Bupleurum Scorzonerifolium Willd

Bupleurum scorzonerifolium Wil Id.is a perennial herb of the genus Bupleurum in the Umbelliferae family.It is widely distributed in my country’s Liaoning,Heilongjiang,Hebei and Jilin provinces and it has a medicinal history of more than2,000 years in China,Its roots have been used as a Chinese herbal medicine for the treatment of fever,depressed liver and Qi obstructiondistending,pain in the chest.Modern pharmacological studies have shown that Bupleurum has antipyretic,analgesic,anti-inflammatory,immune enhancement,hepatoprotective and choleretic,antibacterial,antiviral and antitumor effects.At present,the compounds isolated from Bupleurum include triterpene saponins,lignans,flavonoids,coumarins,volatile oils and polyacetylenes.Based on the review of the chemical constituents and pharmacological activities of Bupleurum,a systematic study of the chemical constituents of Bupleurum.Part of the isolated compounds were screened for in vitro anti-breast cancer activity,and for the first time to conduct RNA-seq research on compounds with better activity.In this study,silica gel column chromatography,HP-20 macroporous resin colu-mn chromatography,ODS column chromatography,HPLC and other chromatograp-hic methods are used to analyze the nbutanol layer of the 70%ethanol extract from t-he roots of B.scorzonerifolium.The extract was systematically separated,and a total of 18 compounds were obtained.The struct-ures of all the compounds were identified through physical and chemical properties and a variety of spectroscopy methods,incl-uding 9 lignans,8 saponins,and 1 oxylip-ins acetals.These compounds are:dextrobu-rsehernin（1）,bursehernin（2）,dehydrodiconiferyl alcohol-4-O-β-D-glucopyranoside（3）,kaerophyllin（4）,isokaerophyllin（5）,chinensin（6）,chinensinaphthol（7）,7S,8S-4,7,9,9’-tetrahydroxy-3,2’-dimethoxy-4’-allyl-8,4’-oxyneolignan（8）,7R,8S-4,7,9,9’-tetrahydroxy-3,2’-dimethoxy-4’-allyl-8,4’-oxyneoli-gnan（9）,solacetal C（10）,2’-O-ac-etylsaikosaponin-b₂（11）,saikosaponin-a（12）,saikosaponin-d（13）,bupieurosideⅢ（14）,prosaikogenin F（15）,saikosaponin-b₄（16）,saikosaponin-b₃（17）,saikosaponin-b₂（18）,among them,compounds 11 is new compounds.In this paper,two breast cancer cell lines,SK-BR-3 and MCF-7,were selected,and the anti-tumor activities of the 1-18 compounds isolated by MTT method were stu-died.The test results show that compared with lignan compounds and oxylipin acetals compounds,saponin compounds exhibit a better inhibitory effect on SK-BR-3and MCF-7 cells.Among them,the inhibitory effect of compound 13 on the two breast cancer cells was close to that of the positive control drug.RNA-seq analysis of compound 13 and GO enrichment analysis showed that the use of saikosaponin-d to treat breast cancer may be through a synergistic effect of biological processes,mole-cular functions,and cell composition.KEGG analysis of RNA-seq sequencing results showed that a variety of pathways related to breast cancer are closely related to the mechanism of action of breast cancer.