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C21 steroid saponin compound in marsdenia tenacissima stem, and applications thereof

A compound, steroidal saponin technology, applied in the field of natural product compound pharmaceuticals, can solve the problems of slow splitting rate, late diffusion and transfer, etc.

Active Publication Date: 2019-02-26
MINZU UNIVERSITY OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Compared with small cell lung cancer (SCLC), NSCLC cells have a longer growth cycle, slower division rate, and relatively late spread and metastasis

Method used

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  • C21 steroid saponin compound in marsdenia tenacissima stem, and applications thereof
  • C21 steroid saponin compound in marsdenia tenacissima stem, and applications thereof
  • C21 steroid saponin compound in marsdenia tenacissima stem, and applications thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0018] Effect of Example 1 on Cell Migration and Invasion

[0019] experimental method

[0020] Migration assay: A549 cells were mixed at 1×10 per well 6 cells were seeded into a 6-well plate, and when the A549 cells adhered to 90% density, use a 20 μl pipette tip to scrape off a monolayer of A549 cells in the middle of each well, wash three times with PBS at 28 μM , 44 μM, 29 μM, and 47 μM were added to the medium containing TGT-7, TGT-9, TGT-13, and TGT-15 for 36 hours, and the control group was supplemented with DMEM medium. Then three points of each wound were selected, and the distance between adjacent wounds was calculated with Image J.

[0021] Invasion test: place the Matrigel gel overnight at 4°C, and after thawing, add serum-free medium to a final concentration of 1 mg / mL to prepare the Matrigel gel. Add 100 µL of prepared Matrigel vertically to the bottom of the upper chamber and 600 µL per well containing 10% FBS to the lower chamber. A549 cells were resuspende...

Embodiment 2

[0023] Example 2 Cell cycle analysis

[0024] experimental method

[0025] Cell cycle analysis was performed using flow cytometry. A549 cells were treated with TGT-7 (28 μM), TGT-9 (44 μM), TGT-13 (29 μM) and TGT-15 (47 μM) for 24 hours, collected and washed twice with ice-cold PBS. The deposited cells were shaken with 70% ethanol and suspended in a -20°C freezer for more than 24 hours.

[0026] After the cells were fixed, they were placed on a centrifuge and centrifuged at 1000 rpm for 5 minutes. Aspirate the ethanol and wash three times with ice-cold PBS. Discard the supernatant. Add 0.5mL PI / RNase to the cells and let stand in the dark for about 15 minutes. Immediately perform cell cycle analysis using flow cytometry.

[0027] Treat A549 cells with TGT-7 (28 μM), TGT-9 (44 μM), TGT-13 (29 μM), TGT-15 (47 μM) for 24 hours, then fix A549 cells and stain with propidium iodide, and the cells are treated with cell flow Cell cycle phase transition of A549 cells was detecte...

Embodiment 3

[0028] Example 3 Cell Apoptosis Analysis

[0029] experimental method

[0030] Cell apoptosis was analyzed using flow cytometry. A549 cells were treated with TGT-7 (28 μM, 56 μM), TGT-9 (44 μM, 88 μM), TGT-13 (29 μM, 58 μM), TGT-15 (47 μM, 94 μM) for 24 h, and then the supernatant was collected in a 15 mL tip In the first centrifuge tube, the adherent cells were trypsinized and separated, collected into corresponding centrifuge tubes, centrifuged at 1000rpm for 5 minutes, aspirated the supernatant and washed twice with 1× binding buffer, and then sequentially according to the manufacturer’s instructions. Stained with Annexin V and PI. Finally, the A549 cells were observed with a fluorescence microscope and analyzed with a flow cytometer.

[0031] Annexin V-FITC / PI double staining method was used to detect apoptotic cells. , 94 μM) after 24 hours of treatment, showed more green fluorescent staining compared to the control group ( image 3 A). In addition, using flow cytom...

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Abstract

The invention relates to a C21 steroid saponin compound in marsdenia tenacissima stem, and applications thereof in preparation of medicines used for preventing and treating lung cancer. It is shown bybiological experiments that the C21 steroid saponin compound in marsdenia tenacissima stem is capable of inhibiting tumor cell migration, maintaining tumor cells at G0 / G1 phases, and releasing Cytochrome c through up-regulating of Bax and down-regulating of Bcl-2 expression, activating Caspase 3 and Caspase 9 to induce tumor cell apoptosis, and is capable of activating caspase 8 at the same time,and inducing cell apoptosis through death receptor pathway. The C21 steroid saponin compound is capable of inhibiting tumor cell migration and invading through down-regulating of MMP-2 and MMP-9 protein expression, the effect is obvious, and lung cancer preventing and treating effect is achieved.

Description

technical field [0001] The invention belongs to the field of natural product compound pharmacy, and relates to C21 steroidal saponin compound and application thereof. Background technique [0002] Lung cancer poses a high threat to human health and quality of life, and has become the main cause of cancer death in recent years. In the past ten years, about 1.6 million people around the world have been diagnosed with lung cancer, and the death toll has exceeded 1.3 million. In addition, lung cancer ranks first in the list of cancers that cause death in men, and is second only to breast cancer in women. The cancer report published in the United States in 2017 pointed out that although the incidence of lung cancer is not the first in the United States, it still occupies the first place in the number of cancer deaths. More than a quarter of cancer deaths are caused by it, and about 85% of them- 90% are non-small cell lung cancer (NSCLC). Compared with small cell lung cancer (SC...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/58A61K31/7048A61P35/00
CPCA61K31/58A61K31/7048A61P35/00A61K2300/00
Inventor 刘同祥徐东伟王绍辉程哲康颜昱胡燕兰何翠薇
Owner MINZU UNIVERSITY OF CHINA
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