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OBJECTIVE：To investigate the activity of lycorine to the in vivo apoptosis of tumor cells in H 22-bearing mice and its mechanism. METHODS ：Kunming mice were inoculated subcutaneously with ascites of H 22 hepatoma mice in the armpit of forelimb to establish solid tumor model. After modeling ，mice were randomly divided into negative control group ，positive control group（hydroxycamptothecin 6 mg/kg），lycorine low-dose ，medium-dose and high-dose groups （10，20，40 mg/kg），with 10 mice in each group. Negative control group was given constant volume of normal saline intragastrically ，and administration groups were given relevant medicine intragastrically ，once a day ，for consecutive 7 days. After last medication ，the weight of tumor was detected and anti-tumor rate was calculated. Ascites tumor model of mice was established by intraperitoneal injection of H 22 hepatoma mice ascites ，and then were grouped with same method and given relevant medicine as above. After last medication ， survival time of mice was recorded and the life prolongation rate was calculated. The early apoptotic rate of tumor cells in mice was detected by flow cytometry. On the basis of normal control group （normal mice without tumor ），the mitochondrial membrane permeability of tumor cells in each group was investigated by Calcein AM staining. The changes of mitochondrial potential were investigated by Rhodamine 123 staining. Colorimetry and Western blot assay were adopted to detect the Caspase-3 activity and expression of apoptosis-related protein （Bcl-2，Bax，Cyt-C and Caspase- 9）. RESULTS ：Compared with negative control UN- group，the tumor weight of positive control group and lycorine PYSCT-2017208） groups were decreased significantly ，while the survival time was significantly prolonged ，and the early apoptotic rate of tumor cells was significantly increased （P＜0.05 or P＜0.01）；the anti-tumor rates were 39.41％ ， 23.36％ ， 36.50％ ， 56.93％，and life prolonga tion rates were 49.23％，29.09％， E-mail：ym913@yahoo.com.cn 50.19％，69.08％. Compared with normal control group ，the mitochondrial membrane permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased，while the mitochondrial membrane potential and Bcl- 2/Bax ratio were decreased significantly （P＜0.05 or P＜0.01）. Compared with negative control group ，mitochondrial membrane permeability and Bcl- 2/Bax ratio were decreased significantly in administration groups ，while mitochondrial permeability ，Caspase-3 protein activity and protein expression of Cyt-C and Caspase- 9 were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ：Lycorine can induce the apoptosis of tumor cells in H22-bearing mice ，the effects of which may be associated with opening mitochondrial membrane permeability transition pore to increase mitochondrial permeability ， decreasing mitochondrial membrane potential and up-regulating the expression of apoptosis-related proteins.
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ABSTRACT Hippeastrum goianum (Ravenna) Meerow, Amaryllidaceae, is an endemic species from the Cerrado, Brazil; there are only few studies about its chemistry or biological activity. This study aimed to investigate the occurrence of lycorine in extracts from in vitro H. goianum plantlets, as well as evaluate a possible inhibition of acetylcholinesterase. The ethanol extract of plantlets produced by in vitro seed germination and micropropagation of bulblets was obtained from seedlings from in vitro germination, while the ethanol extract micropropagtion of bulblets was obtained from a subculture of those seedlings. The presence of lycorine was detected in only in the micropropagation of bulblets. The micropropagation of bulblets was more active than the plantlets produced by in vitro seed germination, with an IC50 of 114.8 ± 0.95 µg/ml and IC50 386.00 ± 0.97 µg/ml, respectively. These results showed that both in vitro germination and micropropagation of H. goianum can lead to the biosynthesis of lycorine. Moreover, the micropropagation led to improved anticholinesterase activity.
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Objective: To explore the key genes and potential therapeutic drugs for osteoarthritis (OA) by bioinformatics.Method: The microarray data GSE55235 was downloaded from the data platform of gene expression omnibus (GEO) and the differentially expressed genes were screened by R language software (3.5.0).Then,the differentially expressed genes were subjected to gene ontology (GO) enrichment analysis and Kyoto encyclopedia of genes and genomes (KEGG) signaling pathway analysis with David online database.The protein-protein interaction was analyzed by String 10.5 online database and visual editing was analyzed by Cytoscape v3.6.1 software.Subnetwork module analysis was utilized by MCODE plugin to screen the core genes in the process of OA.Finally,small molecule drugs with potential treatment for OA were analyzed by connectivity map (CMap) database.Result: A total of 556 differentially expressed genes were screened,among which 252 were up-regulated and 304 were down-regulated.These genes were mainly involved in extracellular matrix (ECM) organization,inflammatory response,cell adhesion,immune response,collagen binding,etc.The analysis of KEGG pathway showed that differential genes were mainly involved in ECM-receptor interaction,phosphatidylinositol 3 kinase-protein kinase B (PI3K/Akt) signaling pathway and osteoclast differentiation.Some genes,such as interleukin-6(IL-6),JUN,vascular endothelial growth factor α(VEGFA),FOS,MYC and early growth response gene-1(EGR-1),activating transcription factor-3(ATF-3),playing critical role in the process of OA were identified by protein-protein interaction.Some potential small molecular drugs for the treatment of OA have also been screened,such as lycorine and anisomycin.Conclusion: The selected key genes may be targets for the diagnosis of OA or potential targets for the treatment of OA,and the selected small molecular drugs can be developed as the key drugs for the treatment of OA.
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Objective: To investigate the inhibitory effect of lycorine on human osteosarcoma 143B cells and its underlying molecular mechanism. Methods: Osteosarcoma 143B cells were treated with 0-8 μmol/L lycorine, respectively. Then the effect of lycorine on the proliferation of 143B cells was detected by crystal violet staining, MTT and colony-forming assay. The migration and invasion abilities of 143B cells were detected by scratch wound healing assay and Transwell assay, respectively. The apoptosis of 143B cells was measured by hoechst 33258 staining. The expression levels of migration-and invasion-related proteins matrix metalloproteinase-7 (MMP-7) and MMP-9, as well as apoptosis-related proteins Bcl-2, Caspase 3 and cleaved Caspase 3 (c-Caspase 3) were detected by Western blotting. Furthermore, the transcription activity of T cell factor/ lymphoid enhancer factor (TCF/LEF) was detected by luciferase reporter gene system. The expression levels of β-catenin and its downstream target molecules Cyclin D1 and c-Myc in Wnt/β-catenin signaling pathway were detected by Western blotting. Results: Lycorine significantly inhibited the proliferation, migration and invasion abilities of osteosarcoma 143B cells, and promoted apoptosis (all P < 0.01). The expression levels of migration-and invasion-related proteins MMP-7 and MMP-9 in 143B cells treated with lycorine were down-regulated remarkably (all P < 0.05), and the expression of apoptosis-related protein Bcl-2 was down-regulated, while the expressions of Caspase 3 and c-Caspase 3 were up-regulated (all P < 0.05). The transcription activity of TCF/LEF was significantly decreased (P < 0.01), and the expression levels of β-catenin, c-Myc and Cyclin D1 were all decreased (all P < 0.05) after lycorine treatment. Conclusion: Lycorine may inhibit the proliferation, migration and invasion of human osteosarcoma 143B cells, and promote apoptosis by blocking the activity of Wnt/β-catenin signaling pathway.
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Objective: To investigate the effect and mechanism of lycorine apoptosis in the human Liver cancer cell line HepG-2. Methods: The inhibitory effect of Lycorine on HepG-2 cell growth was evaluated by the dimethylthiazol tetrazolium assay. Morphological apoptotic changes were characterized using an inverted microscope, Hoechst 33258 fluorescence staining and cell ultrastructure. The rate of cell apoptosis was determined by flow cytometry. Mitochondrial membrane potential and intracellular fluorescent intensity of MMP were detected using laser scanning confocal fluorescence microscopy. Expression of the proteins Bcl-2, Bax, cytochrome C, and Caspase-9 was assessed by western blotting. The activity of Caspase-3 protein was quantified using a Caspase-3 activity kit. Results: Lycorine inhibited the growth of HepG-2 cell line with an IC50 of 5.73 μmol/L. After 48 h lycoris radiata alkali treatment, morphological observation indicated that, the density of growth cell became thin, cell shrinkage and cell growth form an irregular shape, tend to appear apoptotic body. Moreover, with the drug concentration increases, the apoptotic body also gradually increased; Typical apoptosis characteristics of visible cells were observed under transmission electron microscopy. Compared with control group, with the increase of concentration of lycoris radiata alkali dosage, cell apoptosis rate increased, mitochondrial membrane potential decreased and MPTP membrane was open. After 48 h treatment of lycorine, relative activity of Caspase-3 increased in the HepG-2 cells, the expression of cytochrome C, Caspase-9 and Bax on the protein level were upregulated, and the expression of Bcl-2 protein was downregulated, and respectively into concentration dependence relationship, with statistical significance. Conclusion: Lycorine radiata had growth inhibition effect on human liver HepG-2 cells, and induced apoptosis of HepG-2 cells through mitochondrial pathway.
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Recently, studies on Lycoris Herb. type alkaloids received the attention of scholars home and abroad. Lycoris Herb. type alkaloids can be devided into seven types according to their molecular structure, including lycorine type, crinine type, galanthamine type, tazettine type, narciclasine type, lycorenine type, homolycorine type, and montanine type. Researches have shown that Lycoris Herb. type alkaloids possess multiple phamocology activities, such as strong antitumor activity on human breast cancer cell (MCF-7), human leukemia cell (HL-60); and strong inhibitory effect of flu virus, measles virus, polio virus, and SARS virus; Lycorine type and crinine type alkaloids have antimalaria effect; Besides, lycorine type has strong anti-acetylcholinesterase effect. In a word, lycorine type and lycoris type alkaloids carry multiple pharmacological effect and belong to promising substances.
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Objective Study configuration inversion of C-2-hydroxy in lycorine to provide a foundation for semi-synthesis of asiaticumine B .Methods Use instrumental analyses to explore the C-2-hydroxy configuration through oxidation and reduc-tion reactions .Results Accomplished configuration inversion of C-2-hydroxy in lycorine with 49% yield .Conclusion We identified a convenient and efficient method for the configuration inversion of C 2-hydroxy in lycorine .
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AbstractA new lycosinine derivative, 9-O-demethyllycosinine B, was isolated from the native Brazilian Hippeastrum breviflorumHerb., Amaryllidaceae, along with the well-known alkaloids lycosinine B and lycorine. The structure of the new compound was established by physical and spectroscopic methods. 9-O-demethyllycosinine B is the third lycosinine variant identified in the Amaryllidaceae family.