Study On Antitumor Chemical Constituents Of Volatile Compounds From Leaves Of Hippeastrum Vittatum

Hippeastrum vittatum is a monocotyledonous Amaryllidaceae plant of the genus Hippeastrum vittatum,native to the Mexican Caribbean sea and northern Argentina.Hippeastrum vittatum has beautiful flowers and bright colors.Through consulting the literature,we know that Hippeastrum vittatum has medicinal value,among which the chemical substances are rich,including alkaloids,volatile oil,sugar and other substances.The volatile oil in the leaves of Hippeastrum vittatum is rich in content and has various pharmacological activities,and has the effects of resisting bacteria,resisting oxidation,promoting penetration and the like.However,because of the low boiling point and the complicated extraction process,there are few studies on the extraction and anti-tumor activity of volatile oil from Hippeastrum vittatum leaves.Therefore,in this study,the extraction process of volatile oil from Hippeastrum vittatum leaves was optimized and the anti-tumor activity of the extracted volatile oil was studied.Provide theoretical data for large-scale industrial extraction of volatile oil from leaves of Hippeastrum vittatum and provide theoretical basis for future research and development of new antitumor drugs.In this paper,the extraction process of volatile components from Hippeastrum vittatum leaves was optimized.The extraction rate of steam distillation,extraction and distillation extraction were 0.39%,0.89%and 0.59%,respectively.The extraction rate was the highest,and the optimum extraction process was obtained by orthogonal test as follows:the ratio of liquid to material was 2,the extraction temperature was 70?,and the extraction time was 3.5 h.The volatile oils obtained by the three methods were analyzed by gas chromatography and mass spectrometry,and 20 kinds of chemical components with high similarity were obtained by extraction,13 kinds by steam distillation and 14 kinds by distillation extraction.The extraction rate was the highest and the chemical components were the most,so the essential oil needed in subsequent articles was extracted by extraction.In the first half of this paper,after a large number of experiments to screen and optimize the process,in the second half of the Hippeastrum vittatum leaf volatile components of anti-tumor activity screening.In the second half of this thesis,CCK-8 was used to detect the inhibitory effect of total volatile oil in Hippeastrum vittatum leaf on the proliferation of HepG-2 cells.It was found that in the range of 2.5 mg/L-80 mg/L,the inhibitory effect of total volatile oil of Hippeastrum vittatum on proliferation of HepG-2 cells was enhanced with the increase of administration concentration,and the inhibitory effect of total volatile oil of on proliferation of HepG-2 cells was increased.The content of palmitic acid in the total volatile oil of Hippeastrum vittatum was the highest by GC-MS,and the literature showed that palmitic acid also had a variety of pharmacological activities.Therefore,it is speculated that the antitumor activity of volatile compounds in Hippeastrum vittatum leaves is reflected by palmitic acid.therefore,the subject will screen the antitumor activity of palmitic acid in the future.In this study,CCK-8 was used to detect the inhibitory effect of palmitic acid in the total volatile oil of Hippeastrum vittatum leaf on the proliferation of HepG-2 cells.It was found that in the range of 100 umol/L-600 umol/L,the inhibitory effect of total volatile oil from Hippeastrum vittatum on proliferation of HepG-2 cells was enhanced with the increase of administration concentration,and the inhibitory effect of total volatile oil on proliferation of HepG-2 cells was increased.umol/L?According to SPSS data analysis,the IC50 of palmitic acid to HepG-2 cells was determined to be 42 umol/L.umol/LThe determined median lethal dose was 80 umol/L at high concentration and 40 umol/L at low concentration 20 umol/L.The IC50 of half lethal dose of hydroxycamptothecin was 40 umol/L.The HepG-2 cells of liver cancer cultured for 24 h were treated with CCK-8 at three concentrations of high and low,and the morphology of HepG-2 cells was observed after 24 h.After 24 h,the cells in the treatment group were observed under the inverted microscope.with the increase of the concentration,the cells gradually appeared the phenomenon of fuzzy boundary,round shape,broken cell structure,cell fragmentation,and the larger the concentration,the longer the action time,the more obvious the phenomenon;HepG-2 cells of liver cancer after 24 h administration were stained by fluorescence microscope to observe the apoptotic bodies increased in turn,and the cell morphology gradually became irregular.it was observed that with the increase of concentration,more and more nuclei were broken,and with the increase of concentration,diffuse fluorescent halo appeared around the cells.umol/LThe higher the concentration,the more obvious the apoptosis.Finally,the apoptosis rate of HepG-2 cells was detected by flow cytometry.umol/LThe apoptosis rate of positive group was(27.746±0.13)%,that of blank group was(1.174±0.27)%,that of high concentration group was(36.155±0.02)%,that of middle concentration group was(20.316±0.18)%,and that of low concentration group was(5.275±0.11)%.umol/LCompared with the control group,the apoptosis rate increased with the increase of concentration in the administration group,which was statistically significant.In summary,the total volatile oil in Hippeastrum vittatum leaves has a value-added inhibitory effect on HepG-2 cells,and palmitic acid in Hippeastrum vittatum leaves has anti-tumor activity on HepG-2 cells.