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Method for constructing mouse liver cancer model by using aristolochic acid I or combination of aristolochic acid I and carbon tetrachloride

A technology of aristolochic acid and carbon tetrachloride, applied in animal husbandry and other fields

Active Publication Date: 2021-01-19
SHANGHAI JIAO TONG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

But so far there is no experimental evidence that aristolochic acid can cause liver cancer

Method used

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  • Method for constructing mouse liver cancer model by using aristolochic acid I or combination of aristolochic acid I and carbon tetrachloride
  • Method for constructing mouse liver cancer model by using aristolochic acid I or combination of aristolochic acid I and carbon tetrachloride
  • Method for constructing mouse liver cancer model by using aristolochic acid I or combination of aristolochic acid I and carbon tetrachloride

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] Embodiment 1, administering to mice by intraperitoneal injection

[0042] 1. Preparation of aristolochic acid I and carbon tetrachloride solution

[0043] 1.1 the aristolochic acid I solution with phosphate buffered saline (PBS) preparation concentration is 0.25 or 0.5mg / ml, carbon tetrachloride (CCl 4 ) is prepared as a 10% carbon tetrachloride solution with corn oil.

[0044]1.2 Put it in a 65°C water bath and heat for 20 minutes to sterilize.

[0045] 2. For C57 / BL6 wild-type male mice born at 1 week or 2 weeks or male mice with Pten liver-specific knockout (Pten gene with LoxP site mice (Pten f / f ) and Alb-Cre mice were purchased from Jackson Laboratory. Pten f / f Multigenerational mating with Alb-Cre to obtain homozygous Pten liver-specific knockout mice (Pten LKO ). ) carry out different times and different doses of aristolochic acid I administration, and for some wild-type mice wherein aristolochic acid I has been administered, carry out the carbon tetrach...

Embodiment 2

[0047] Embodiment 2, mouse anatomy and tumor statistics

[0048] Respectively select the mice after the treatment of each group in Example 1 for dissection and tumor statistics:

[0049] The mouse was anesthetized with isoflurane, and then the blood was collected from the eyeball. After the blood was allowed to stand at room temperature for 30 minutes, the supernatant was collected at room temperature at 5000 rpm for 30 minutes, aliquoted, and stored at -80°C.

[0050] The mice were killed by neck dissection, and the mice were fixed on the dissecting board for dissection, and the heart, liver, spleen, lung, kidney, brain, testis, ureter, bladder, stomach and tail were taken out, and the organs were observed for abnormalities. For the liver with liver cancer, each focal point was measured with a vernier caliper, and the number of focal points was counted, and the liver and kidney were weighed and photographed. The excised parts of each organ were placed in 4% paraformaldehyd...

Embodiment 3

[0053] Embodiment 3, pathological and histochemical determination of mouse liver cancer

[0054] 1 Tissue dehydration, embedding and sectioning

[0055] 1.1 After the tissue is fixed in 4% paraformaldehyde for 24 hours, dehydrate the tissue according to the following procedure:

[0056] Tap water rinse 10mim→50% ethanol 30min→60% ethanol 30min→70% ethanol 30min→80% ethanol 30min→90% ethanol 30min→95% ethanol 30min→95% ethanol 30min→100% ethanol 30min→100% ethanol 30min→none Water ethanol: xylene (1:1) 10min→xylene 5min→xylene 5min→paraffin 60°C 10min→paraffin 60°C 10min.

[0057] 1.2 Embed the wax-soaked tissue block with a paraffin embedding machine, cool it at -20°C, and store it at -20°C.

[0058] 1.3 Adjust the slice thickness of the paraffin microtome to 5 μm, slice and place in 42°C ddH 2 O to unfold the slices, and when fully stretched without wrinkles, scoop out the slices with an adhesive slide, place the slices on a 37°C oven for overnight, and store them at roo...

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Abstract

The invention discloses a method for constructing a mouse liver cancer model by using aristolochic acid I or combination of the aristolochic acid I and carbon tetrachloride. The method comprises the following steps of performing intraperitoneal injection on a mouse to perform administration of the aristolochic acid I or the combination of the aristolochic acid I and the carbon tetrachloride; and selecting the treated mouse to carry out dissection, tumor statistics and pathohistochemical judgment on the liver cancer of the mouse. The method provides a powerful tool for researching a liver cancer induction mechanism of the aristolochic acid I, mutation and mutation fingerprints caused by the aristolochic acid I in a cancer process and treatment of the liver cancer induced by the aristolochicacid I.

Description

technical field [0001] The present invention relates to the use of chemical carcinogens to establish a mouse liver cancer model, aiming at aristolochic acid I to establish a mouse liver cancer model, in particular to a group of mice using aristolochic acid I or its combination with carbon tetrachloride to build a mouse liver cancer model method. Background technique [0002] In the 1990s, researchers found that aristolochic acid (AA; mainly containing aristolochic acid I or aristolochic acid II) can cause aristolochic acid nephropathy, and then found that aristolochic acid can cause Urothelial carcinoma is listed as a class I human carcinogen by the International Agency for Research on Cancer. Aristolochic acid is a genotoxic chemical carcinogen, and its metabolites can combine with purine bases to form AA-DNA adducts—aristolactam (AL)-DNA adducts (dA-AL and dG-AL), as a specific marker of AA exposure, can cause DNA mutations characterized by A>T transversions, and the ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01K67/02
CPCA01K67/02
Inventor 韩泽广路兆宁
Owner SHANGHAI JIAO TONG UNIV
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