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New medical application for cysteine sulfinate

A cysteine ​​sulfinic acid and drug technology, applied in the field of biomedicine, can solve problems such as unclear biological function of cysteine ​​sulfinic acid

Inactive Publication Date: 2019-04-26
SOUTHWEST UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

The biological function of cystesulfinic acid, the direct catalytic product of CDO, is still unclear

Method used

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  • New medical application for cysteine sulfinate
  • New medical application for cysteine sulfinate
  • New medical application for cysteine sulfinate

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] In order to reveal the regulatory effects of CDO and CSAD on liver damage repair and liver fibrosis, we used CRISPR / Cas9 technology to set two reverse shortenings in the coding region of the first exon of the CDO gene and the second exon of the CSAD gene. The sgRNA target ( figure 2 A. image 3 A) Cooperate with SpCas9n (D10A) nicking enzyme to avoid off-target occurrence. Use T7 transcription kit (mMessage Machine T7kit, Life Company) to synthesize SpCas9n gene mRNA, use T7 high-efficiency transcription kit (Hiscribe TM T7High yield RNA synthesis Kit, NEB) synthesizes sgRNA corresponding to two genes. Microinjection of SpCas9n mRNA (60ng / μL) and two sgRNAs (20ng / μL) through mouse fertilized eggs and embryo transfer were used to obtain CDO ( figure 2 B) and CSAD ( image 3 B) Frameshift mutant mice. After expansion, individuals with homozygous mutations (CDO KO and CSAD KO) were obtained, and the CDO and CSAD knockout mice were verified by Western Blot to have the C...

Embodiment 2

[0040] The 8-week-old CDO gene knockout (CDO KO), CSAD gene knockout (CSAD KO) and wild-type (WT) male mice were selected respectively to undergo CCl 4 Treatment of liver damage. According to the concentration of 0.2% CCL 4 Dissolved in sesame oil, the injection dose is 10ml / kg body weight, the injection frequency is intraperitoneal injection every 2 days, and liver samples are collected after 2 weeks of treatment (day 14). Three hours before sampling, the mice were intraperitoneally injected with BrdU solution (10 mg / ml) at a dose of 50 mg / kg body weight. The mice were sacrificed by severed neck, and fresh liver tissue was quickly collected and placed in 4% paraformaldehyde fixative (pH 7.4) for 6 hours at room temperature. The liver tissues were embedded in paraffin, sectioned, baked, deparaffinized, and analyzed by HE staining, Masson staining, α-SMA immunostaining, Sirius scarlet staining, BrdU immunostaining and TUNEL analysis after entering water.

[0041] According to the...

Embodiment 3

[0049] In order to further verify the function of CDO's direct catalytic product CSA against liver oxidative damage and inhibit liver fibrosis, CDO KO mice were subjected to CCl 4 Injection treatment (the method is the same as in Example 2), and at the same time, 0.9% NaCl (10ml / kg body weight) (control group) or 1% CSA solution (100mg / kg body weight) is injected daily according to the injection volume of 10ml / kg body weight (treatment group). The sampling and slice preparation methods were the same as in Example 2, and then HE staining, Masson staining, Sirius scarlet staining, BrdU immunostaining and TUNEL analysis were performed.

[0050] The results of HE staining showed that the central vein area and portal area of ​​the NaCl control mice showed a large number of immune cell infiltration and balloon edema degeneration, while CDO KO mice had liver immune cell infiltration, balloon edema degeneration, and eosinophilia after CSA supplementation. Damages such as degenerated bodi...

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PUM

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Abstract

The invention relates to applications of cysteine sulfinate in preparing drugs for preventing or treating liver impairment, in preparing drugs for preventing or treating oxidative damage to liver, inpreparing drugs for preventing or treating liver fibrosis, in preparing drugs for preventing or treating hepatic diseases caused by liver fibrosis and in preparing drugs for preventing or treating hepatic diseases caused by oxidative damage. In the invention, a CC14 induced liver injury model, mutual verification on the basis of analysis results of liver tissue injury, activation degree of hepaticstellate cells and hepatic fibrosis of a CDO gene knockout rat and a CSAD gene knockout rat and a CSA compensation test prove that an intermediate product CSA of cysteine oxidative metabolism pathwayplays an important role in repairing liver injury and regulating activation process of hepatic stellate cells and is an effective drug for preventing and treating hepatic diseases.

Description

Technical field [0001] The invention relates to the field of biomedicine, in particular to the new use of cysteine ​​sulfinic acid in the preparation of drugs for preventing or treating liver injury. Background technique [0002] The liver is the largest digestive gland in the body and the core organ for sugar, lipid, and protein metabolism. At the same time, the liver is also responsible for the biotransformation of drugs and endogenous poisons. The physiological functions and physiological characteristics of the liver make it vulnerable to physical, chemical (such as alcohol, mycotoxin and other toxic substances) and biological (such as virus, parasitic infection, etc.) damage, and many types of stimulation can cause it Liver damage, long-term liver damage triggers liver fibrosis, while liver fibrosis is too strong and lasts too long, it will evolve into cirrhosis and liver failure. [0003] Liver fibrosis is widespread worldwide and is the main cause of most liver diseases and ...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/185A61P1/16
CPCA61K31/185A61P1/16
Inventor 赵建军马兴宇刘安芳韩玉竹马启旺叶国根周乐乔冰珂卢晓健韩凌云
Owner SOUTHWEST UNIVERSITY
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