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Application of theabrownin in preparation of medicine for treating liver cancer

A technology for the treatment of liver cancer and theabrownin, which is used in drug combinations, pharmaceutical formulations, anti-tumor drugs, etc., can solve the problem of unclear mechanism of action and its impact, and achieve the effect of inhibiting tumor growth and promoting apoptosis

Active Publication Date: 2022-03-11
杭州茗褐生物科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] However, the relationship between theabrownin and JNK signaling pathway and its role in inhibiting liver cancer are still unknown, and its mechanism of action and its impact are still unclear, and follow-up studies are needed

Method used

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  • Application of theabrownin in preparation of medicine for treating liver cancer
  • Application of theabrownin in preparation of medicine for treating liver cancer
  • Application of theabrownin in preparation of medicine for treating liver cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0024] Example 1: Effect of theabrownin on the proliferation of Huh7 cells

[0025] HepG2, SK-hep1, and Huh7 cells were treated with theabrownin, and the cell viability after 24 and 48 hours was observed. cells in 8 x 10 3 Cells / well were seeded on a 96-well plate and cultured in 200 µl medium for 24 hours. Add 200µl of theabrownin at different concentrations, culture in the incubator for 24 hours and 48 hours respectively, suck off the supernatant, add 20µl of MTT solution (5.0mg / ml) to each well and incubate at 37°C for 4 Hour. Carefully suck out the culture medium in the wells, then add 150 µl DMSO to each well, shake on a shaker at low speed for 10 minutes to fully dissolve the crystals, and measure the optical density (OD) value at 490 nm with a microplate reader. Inhibition rate (IR, %)=[1−(theabrownin treated OD / untreated OD)]*100%. 100 mg / ml, 200 mg / ml and 300 mg / ml were used as the low, medium and high doses of theabrownin for the next experiment.

[0026] The re...

Embodiment 2

[0027] Example 2: Effect of theabrownin on apoptosis of Huh7 cells

[0028] DAPI staining and flow cytometry were used to observe the apoptosis morphology of Huh7 cells after theabrownin treatment. cells in 8 x 10 3 The density of each well was inoculated on a 24-well plate, and treated with low, medium, and high doses of theabrownin for 24 hours, and the control group was added with a culture medium without theabrownin, and cultured for 24 hours. Cells were fixed with 4% paraformaldehyde for 30 minutes at room temperature, and washed with PBS buffer solution for 3 times, 5 minutes each time. Permeabilize with PBS containing 0.5% Triton-X-100 at room temperature for 30 minutes, choose PBS buffer solution to wash 3 times, 5 minutes each time. Add DAPI staining solution for 4 minutes at room temperature, choose PBS buffer solution to wash 3 times, 5 minutes each time. Unstained and stained cells were observed using a fluorescence microscope. Five funnels in each group were u...

Embodiment 3

[0031] Example 3: Effect of theabrownin on apoptosis of SK-hep1 cells

[0032]DAPI staining and flow cytometry were used to observe the apoptotic morphology of SK-hep1 cells after theabrownin treatment.

[0033] The result is as image 3 As shown in A, with the increase of theabrownin concentration, more and more apoptotic cells appeared chromatin condensation, nuclear pyknosis and apoptotic bodies (arrows). Such as image 3 As shown in B, as the concentration of theabrownin increased, the early and late apoptosis rates of SK-hep1 cells increased significantly, indicating that theabrownin induced apoptosis of SK-hep1 cells in a dose-dependent manner.

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Abstract

The application of theabrownin in the preparation of drugs for treating liver cancer belongs to the technical field of theabrownin use. Specifically include: the application of theabrownin in the preparation of drugs for treating liver cancer, theabrownin promotes the apoptosis of hepatocellular carcinoma Huh7 cells by activating the JNK signaling pathway, and inhibits the growth of liver cancer xenografts. The present invention proves through various tests that theabrownin has significant anti-proliferation and pro-apoptotic effects on Huh7 cells, and has dose-dependent effects. Theabrownin promotes the apoptosis of Huh7 cells and inhibits the growth of zebrafish liver cancer xenografts by activating the JNK signaling pathway. Theabrownin can inhibit tumor by 48.1%. Moreover, theabrownin regulates downstream apoptosis-related genes by activating the P53 pathway PUMA, NOXA, Bax, and Bcl‑2 and so on to promote the apoptosis of P53 wild Sk‑hep1 cells.

Description

technical field [0001] The invention belongs to the technical field of theabrownin application, and in particular relates to the application of theabrownin in the preparation of drugs for treating liver cancer. Background technique [0002] Theabrownin (TB) is a complex natural polymer extracted from tea leaves that is soluble in water but insoluble in ethyl acetate and n-butanol. It is composed of polyphenols such as theaflavin and thearubigin Derived from further oxidation, it is naturally present in black tea, green tea and black tea. Theabrownin is of great benefit to improving the comprehensive metabolic balance of the human body. It has the effects of lowering blood sugar, blood lipids, blood pressure, and uric acid. It is also a promising anticancer candidate substance. [0003] Liver cancer is a malignant tumor of the liver, which can be divided into two categories: primary and secondary. Primary malignant tumors of the liver originate from the epithelial or mesenc...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K36/82A61P35/00A61K127/00
CPCA61K36/82A61P35/00
Inventor 张进
Owner 杭州茗褐生物科技有限公司
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