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Anti-hepatic fibrosis medical application of dammarane type sapogenin

An anti-liver fibrosis, total saponin technology, applied in the field of medicine, can solve the problems of not reaching the expected effect, complex formation mechanism of liver fibrosis and the like

Inactive Publication Date: 2015-06-03
SHENYANG PHARMA UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The formation mechanism of liver fibrosis is very complicated. Many in vitro and animal experiments have proved effective treatment methods, but they have not achieved the expected effect after being used clinically.

Method used

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  • Anti-hepatic fibrosis medical application of dammarane type sapogenin
  • Anti-hepatic fibrosis medical application of dammarane type sapogenin
  • Anti-hepatic fibrosis medical application of dammarane type sapogenin

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] Embodiment 1: MTT cell viability experiment

[0018] Take t-HSC / Cl-6 cells (hepatic stellate cells) at 1×10 4 The density per well was seeded on a 96-well plate, cultured at 37°C for 24 hours with a medium (complete medium) containing 10% fetal bovine serum, and then added with 25-OCH 3 Cells were stimulated by 12 different compounds including -PPD and 25-OH-PPD. At the same time, normal human intestinal wall cells were treated with 1×10 4 The density of each well was seeded on a 96-well plate and given different concentrations of 25-OCH 3 -PPD and 25-OH-PPD to observe their effects on normal intestinal cells like figure 2 . 5 mg / mL MTT was dissolved in PBS, sterilized by filtration, and 100 μL DMSO was added to fully dissolve the blue-purple formazan precipitate, and the absorbance value of each well at 540 nm was read on an enzyme label detector to observe the effect of the drug on cytotoxicity. The result is as figure 1 Shown, 25-OCH 3 -PPD and 25-...

Embodiment 2

[0019] Embodiment 2: GSH glutathione detection

[0020] Add different concentrations of 25-OCH to hepatic stellate cells 3 -PPD solution, after 12 h, take the mixed solution and centrifuge at 2000 rpm for 10 min at 4°C. The supernatant was deproteinized and mixed with the test solution, and the absorbance of the sample was measured at 400 nm. The result is as image 3 Increased over time, 25-OCH 3 -PPD significantly increased intracellular GSH concentration in a concentration-dependent manner.

Embodiment 3

[0021] Example 3: 25-OCH 3 -PPD Inhibiting the expression of survivin and Bcl-2 protein

[0022] 25-OCH 3 -PPD (3.125, 6.25, 12.5 μM) can significantly induce and activate the caspase-3 activation fragment in HSC-T6 by inhibiting the expression of survivin. Among them, 25-OCH 3 -PPD (6.25 μM, 12.5 μM, 25 μM) increased the activity of caspase-3 by 8-9 times, respectively, and decreased the expression of survivin by 1.6%, 28.4% and 42.2%, as shown in Figure 4 -A . Bcl-2 protein was treated with 12.5 μM and 25 μM 25-OCH 3 - Significantly lower after PPD. show that 25-OCH 3 -PPD can reduce the expression of Bcl-2 protein in hepatic stellate cells through the mitochondrial pathway, and alleviate cell apoptosis, such as Figure 4 -B .

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Abstract

The invention belongs to the technical field of medicines and relates to an anti-hepatic fibrosis medical application of dammarane type sapogenin 20(R)-25-methoxy-dammarane-3beta,12beta,20-triol and 20(R)-25-hydroxyl-dammarane-3beta,12beta,20-triol (which are called as 25-OCH3-PPD and 25-OH-PPD for short). Study shows that 25-OCH3-PPD and 25-OH-PPD can be used for reducing expression of alpha-SMA, TGF-beta and TIMP-1, improving caspase-3 level and increasing signal paths for NF-kappaB redistribution and generation of Bcl-2 proteins in hepatic stellate cells, and the hepatic stellate cells are reversed to be in a resting state, so that hepatic fibrosis is reversed, and 25-OCH3-PPD and 25-OH-PPD can be used for preparing a medicine for treating the hepatic fibrosis.

Description

technical field [0001] The invention relates to the technical field of medicine, and relates to 25-OCH 3 -Application of PPD and 25-OH-PPD in the preparation of anti-hepatic fibrosis drugs. Background technique [0002] Hepatic fibrosis is one of the diseases with the greatest impact in the pathological changes of the liver, and it is a pathological change common to persistent liver injury. my country is a high-incidence area of ​​hepatitis virus infection, and the probability of inducing liver fibrosis is very high. Hepatic fibrosis is an intermediate link in the development of chronic liver disease to cirrhosis. If it is not treated in time, it will progress to cirrhosis, and the final deterioration can only be maintained by liver transplantation. The short-term curative effect is sure and clear, but the long-term curative effect is affected by the immune system The restriction of rejection is not optimistic, and it is affected by the source of liver and recipients of li...

Claims

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Application Information

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IPC IPC(8): A61K31/575A61P1/16A61P35/00
Inventor 赵余庆南极星吴艳玲
Owner SHENYANG PHARMA UNIVERSITY
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