Study On The Chemical Constituents In Hypericum Sampsonii And Their Anti-tumour And Anti-inflammatory Activity

Hypericum sampsonii belongs to the hypericum plant family of Guttiferae,and it is also known as Wang Buliu Xing,Liu Jinu,and leafy grass,ect.It is a perennial herb.The plant is widely distributed in the north and south provinces of China,of which Hubei and Jiangsu,Zhejiang and Sichuan are more common,it is a unique species in China.The folk uses whole grass as medicine,which has the effects of cooling blood to stop bleeding,clearing heat and detoxifying,promoting blood circulation,regulating menstruation,and removing wind and collaterals.It can be used to treat vomiting blood,hemoptysis,traumatic bleeding,enteritis,dysentery,dysentery,bloated and poisonous,burns,snake bites,irregular menstruation,rheumatism.H.sampsonii is a traditional Chinese herbal medicine.The results of modern pharmacological studies show that Hypericum plants have various biological activities such as antidepressant,antibacterial,antiviral and antitumor.At present,according to literature reports,there have been some researches on the chemical composition of the plant at home and abroad.In order to further explore the material basis for the activity of H.sampsonii,we will systematically study the chemical components of the palnt from Hubei and its antitumor and anti-inflammatory activities in vitro.Objective:Extraction,purification,and structural identification of H.sampsonii,and preliminary research on the antitumor and anti-inflammatory activity of the isolated single compound,in order to provide basic data for related research on the efficacy of herbs.Methods:Silica gel column chromatography,thin-layer chromatography,C18 column,Sephadex LH-20 gel column,HPLC and other separation methods were used to isolate and identify 25 compounds from H.sampsonii.Their chemical structures were determined by 1H-NMR,13C-NMR and mass spectrometry.MTT was used to determine the Et OH extract of H.sampsonii and 25 monomer compounds against 5human tumor cell lines:human melanoma cells?A 375?,human gastric cancer?SGC-7901?,Human breast cancer?MDA-MB-231?,human cervical cancer cells?Si Ha?,and human bone marrow neuroblastoma cells?SHSY-5Y?.Griess reagent method was used to determine the effect of monomer compounds on NO release from RAW264.7macrophages stimulated by LPS,so as to determine the anti-inflammatory activity of monomer compoundsResults:Twenty-five compounds were isolated from the Et OH extract of H.sampsonii.Of them,compounds 3,4,6,7,10-14,17,19,22 and 23 were reported in H.sampsonii for the first time.HPLC analysis showed that this study has almost completed the separation and identification of most of the plant's components,which is an important basis for the next step of activity testing.MTT assay revealed that compounds 1,2,5-14,15,17,18,20,21,22 and 25 had selective cytotoxic activities(IC₅₀:7.52-158.90?M)against tested cancer cells,in which compound 5,6,13 and 14 displayed high activities against A375,MDA-MB-231,Si Ha and SHSY-5Y.In LPS-stimulated RAW 264.7macrophages assay,most compounds?1-3,5-23,25?showed considerable high anti-inflammatory activities(IC₅₀:10.59-41.69?M),in which compounds 5,6,13,14,and 15 exhibited high anti-inflammatory activities with IC₅₀values of 12.39,14.11,17.23,13.05,and 10.59?M,respectively.Conclusion:This study further enriched the research on the chemical constituents of H.sampsonii,and showed that quinones and phloroglucinols may be the active basis for the antitumor and anti-inflammatory effects of H.sampsonii.