The Chemical Constituents And Bioactivity Of The Ether Extract Of Tetrastigma Hemsleyanum Diels & Gilg.

The separation of monomeric chemical components and their activity research is a topic in natural medicine research.Many monomeric compounds have been isolated from the traditional Chinese medicine,but there are only few drug molecules used in clinical applications,and traditional Chinese medicine preparations are more used in clinical practice.However,the separation of monomeric compounds is still of great value for the creation of new drugs and for mining the mechanism of action of traditional Chinese medicines.Tetrastigma hemsleyanum Diels&Gilg.,also known as Sanyeqing,is widely used in Zhejiang and Fujian.Previous studies on the activity of Tetrastigma hemsleyanum Diels&Gilg.were mostly focused on the activity of mixtures.In Zhejiang,there is a custom of taking Tetrastigma hemsleyanum Diels&Gilg.at it’s low temperature state.Our previous experiments showed that the anti-inflammatory effect of Tetrastigma hemsleyanum Diels&Gilg.ethyl ether extract is more obvious than that of ethanol extract.Therefore,in this study,ethyl ether was used as solvent and combined with lowtemperature ultrasonic extraction technology to extract and separate the components and activities of Tetrastigma hemsleyanum Diels&Gilg..The main results are as follows:Ethyl ether was used to extract the components from the dried root powder of Tetrastigma hemsleyanum Diels&Gilg.,and divide the ethyl ether extract into several components by silica gel chromatography column,recrystallization and other methods,then carry out separation and purification.For monomeric compounds,the structure of the compound is determined by combining the physicochemical properties,mass spectrometry and nuclear magnetic resonance spectral data of the compound.The results showed that a total of 4 compounds were isolated and identified as 2,6-di-tert-butyl-pcresol(1),β-Sitosteryl linoleate(2),Palmitic acid(3),β-Sitosterol(4).Among them,compounds(1)and(2)were isolated from Tetrastigma hemsleyanum Diels&Gilg.for the first time.The anti-inflammatory activity of the isolated crude fractions and monomeric compounds was analyzed by bacterial lipopolysaccharide(LPS)-induced human microvascular endothelial cells(HMEC-1)inflammation model,and MTT(3-[4,5dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide)method was used to determine the toxic effects of crude fractions and monomeric compounds on HMEC-1 cells.It was found that only 2,6-di-tert-butyl-p-cresol(1)as a monomeric compound exhibited antiinflammatory activity,but compound(1)was toxic to HMEC-1 cells with IC50 of 0.204 mmol/L,so it was considered that its toxicity caused the anti-inflammatory effect.The crude fractions have good anti-inflammatory activity and show no obvious toxicity.It can be considered that the anti-inflammatory effect of Tetrastigma hemsleyanum Diels&Gilg.is more direct and obvious in the multi-component mixture.The anti-tumor activity of monomeric compounds on Human Non-Small Cell Lung Cancer Cells A549 and Human Colorectal Cells HT29 was detected by MTT assay.The results showed that compound(1)significantly inhibited the proliferation of A549 and HT29 with IC50 of 1.01 mmol/L and 0.77 mmol/L,respectively.The proliferation inhibitory activity of compound(3)on A549 and HT29 was also obvious,with IC50 of 0.199 mmol/L and 2.34 mmol/L,respectively.It verifies the scientificity of the use of Tetrastigma hemsleyanum Diels&Gilg.as an anti-tumor drug in the folk.The preservative group was uesd as control,the powder and the ethanol extract as active ingredient additives,two toothpastes containing Tetrastigma hemsleyanum Diels&Gilg.were prepared.And then treated at 4℃,20℃,and 37℃ for 90 d.Check out the changes of those toothpastes.The results showed that after 90 days of treatment at different temperature,the appearance of the prepared toothpaste was no different from that of the new toothpastes.