ABSTRACT
To investigate the inhibitory effects of two xanthone compounds, 1-hydroxy-2,3,4,8-4 methoxy xanthone(here in after referred to as Fr15) and 1-hydroxy-2,3,4,6-4 methoxy xanthone(here in after referred to as Fr17), on the proliferation of hepatocellular carcinoma cells HepG2, and to further investigate their mechanism in combination with transcriptomics. Cell counting was used to detect the effects of two kinds of xanthone compounds Fr15 and Fr17(0, 0.03, 0.15, 0.3 mmoL·L~(-1)) on the proliferation of HepG2 cells; the effects of the two compounds Fr15 and Fr17 on HepG2 cell cycle were detected by flow cytometry; the changes of autophagosomes count in cells were observed under fluorescence microscope; the expression of autophagy marker proteins autophagy marker proteins SQSTM 1(p62) and microtubule associated protein 1 light chain 3 Ⅰ/Ⅱ(LC3 Ⅰ/Ⅱ) in the cells was detected by Western blot; the differentially expressed genes between the control group and the experimental group were analyzed by RNA-seq transcriptome sequencing; qRT-PCR was used to verify the differentially expressed genes in sequencing. The results showed that compounds Fr15 and Fr17 inhibited the proliferation of HepG2 cells with the increase of drug concentration and time. Flow cytometry showed that compounds Fr15 and Fr17 had little effect on HepG2 cell cycle. Fluorescence microscopy results showed that the number of autophagosomes in cells increased with the increase of drug concentration. Western blot showed that the expression of p62 protein was decreased and the expression of LC3-Ⅱ protein was significantly increased after drug addition. The results of RNA sequencing showed that 26 102 and 52 351 differentially expressed genes were obtained in Fr15 and Fr17 respectively. Analysis of KEGG showed that drug treatment had a great effect on autophagy pathway. qRT-PCR verified that 6 up-regulated genes were related to autophagy, and their trend was consis-tent with sequencing results, where all 6 genes showed an up-regulated trend. Two xanthone compounds Fr15 and Fr17 may inhibit proliferation of HepG2 cells by inducing autophagy.
Subject(s)
Apoptosis , Autophagy , Cell Cycle , Hep G2 Cells , XanthonesABSTRACT
OBJECTIVE： To study the phylogenetic relationships of 3 basic plants of Tibetan medicine “Dida”， such as Swertia puricea， Wertia mileensis， Halenia elliptica. METHODS： ISSR technology was used for PCR amplification of 9 samples of S. puricea （ZT-1 to ZT-5 from Gantongsi in Dali Cangshan， ZC-1 to ZC-4 from Binchuan county of Dali）， 2 samples of W. mileensis （QYD-1 to QYD-2） and 2 samples of H. elliptica （HM-1 to HM-2）. Using DNA genome of S. puricea as template， 8 primers were screened and used for PCR reaction. The PCR amplification products were read by hand， the original data matrix was established， and the polymorphic band ratio was calculated. At the same time， genetic similarity coefficient was calculated by using NTSYS 2.1 software， and UPGMA method was used to draw cluster diagram. RESULTS： A total of 113 clear and identifiable amplification product bands were obtained by 8 ISSR primers. The rate of polymorphic site was 100％. The genetic similarity coefficients for totally 13 samples of S. puricea， W. mileensis and H. elliptica ranged 0.301-0.500. Intraspecific genetic similarity coefficients for 9 samples of S. puricea ranged from 0.752 to 0.929. The cluster analysis showed， when the range line was 0.410， 13 samples could be divided into three groups， i.e. S. puricea， W. mileensis， H. elliptica； when the range line was 0.780， 9 samples of S. purpurea could be divided into 2 subgroups， one of which was only sample ZT-1 collected from Gantongsi in Cangshan， and the other contained the remaining 8 samples. CONCLUSIONS： ISSR technology can be used to identify S. punicea， S. glabra and H. elliptica at the molecular level. S. punicea has some genetic relationship with S. glabra and H. elliptica， but the genetic relationship is relatively distant and the genetic difference is large. S. punicea from two different locations in Dali area has little genetic difference and close relationship， but it shows abundant genetic diversity.
ABSTRACT
Objective To investigate the protective effects of ethanol-extract of Halenia elliptica D. Don on chemical hepatic injury in mice. Methods Mouse models of chemical hepatic injury were induced by intraperitoneal injection of 0.12 %CCL4. Extract of Halenia elliptica D. Don by alcohol was administered,and serum ALT,AST activities and liver glycogen level were measured in mice. Results Compared with the models,the enzyme activities of ALT and AST were significantly reduced and the content of liver glycogen was significantly increased in the ethanol-ex tract of Halenia elliptica groups. Conclusion It is indicated that ethanol-extract of Halenia elliptica D. Don has an effect in protecting the liver.
ABSTRACT
Object To separate and characterize the chemical constituents of a plateau plant Halenia elliptica D Don Methods Elemental analysis (EA), 1HNMR and 13 CNMR, MS, FTIR and UV spectrometry, as well as DSC were employed Results Two needle shaped crystal chemical constituents obtained from H elliptica were confirmed to be 1 hydroxy 3, 7, 8 trimethoxyxanthone and 1, 7 dihydroxy 3, 8 dimethoxyxanthone, respectively Conclusion This is the first time for these two chemical constituents to be separated from this Tibetan medicinal plant