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Application of insulin-like growth factor 2 recombinant protein in preparation of medicines for treating ulcerative colitis

A technology for ulcerative colitis and growth factors, which is applied in the field of medicine, can solve the problems of unreported application of IGF2 recombinant protein, and achieve the effect of preventing ulcerative colitis, reducing the level of inflammatory factors, and enhancing the expression

Inactive Publication Date: 2021-03-12
THE FIRST AFFILIATED HOSPITAL HENGYANG MEDICAL SCHOOL UNIV OF SOUTH CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] At present, there is no report on the application of IGF2 recombinant protein in the treatment of ulcerative colitis

Method used

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  • Application of insulin-like growth factor 2 recombinant protein in preparation of medicines for treating ulcerative colitis
  • Application of insulin-like growth factor 2 recombinant protein in preparation of medicines for treating ulcerative colitis
  • Application of insulin-like growth factor 2 recombinant protein in preparation of medicines for treating ulcerative colitis

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] The animals selected in this experiment are 8-week-old C57BL / 6 male mice, weighing about 25 grams, purchased from Hunan Slake Jingda, license number: SCXK (Xiang) 2019-0004, and the mice were placed under artificial day and night cycle lighting for 12 hours Raised in the environment, free access to food and water. C57BL / 6 male mice were randomly divided into sterile water+BSA control group (8 rats), DSS+BSA model group (8 rats), DSS+IGF2 recombinant protein administration group (8 rats). The control group was intraperitoneally injected with BSA (15 μg / kg) once a day and drank sterilized water freely; the model group was injected intraperitoneally with BSA (15 μg / kg) once a day and drank 5% dextran sulfate (DSS) freely; IGF2 recombinant protein (15 μg / kg) was injected intraperitoneally once a day, and 5% DSS was freely drunk. After 7 days of administration, the mice were anesthetized by intraperitoneal injection of 4% chloral hydrate. After the anesthesia was complete, ...

Embodiment 2

[0039] The sources of experimental animals, feeding conditions and dosage regimens refer to Example 1. After the mice were anesthetized, colonic tissues were taken, photographed, and fixed in 4% paraformaldehyde for 24 hours for tissue HE staining.

[0040] Colon tissue HE staining steps:

[0041] (1) After dehydrating the fixed colonic tissue according to 30% alcohol-50% alcohol, 70% alcohol-80% alcohol-90% alcohol-95% alcohol-100% alcohol-100% alcohol (1h each), Soak in xylene for 2 hours to make it transparent, then soak it in paraffin liquid for 1 hour, and use a paraffin embedding machine to make paraffin blocks.

[0042] (2) Cut the paraffin block into 5 μm paraffin sections using a paraffin microtome.

[0043] (3) After the paraffin sections were dewaxed by soaking in xylene for 1 h, the gradient hydration was carried out according to 100% alcohol-100% alcohol-90% alcohol-80% alcohol-70% alcohol-distilled water (each 1 min).

[0044] (4) Add hematoxylin staining solu...

Embodiment 3

[0054] The sources of experimental animals, feeding conditions and dosage regimens refer to Example 1. After the mice were anesthetized, the colon tissues were stored at -80°C for Real-time PCR detection.

[0055] Colon tissue Real-time PCR steps:

[0056] (1) RNA extraction and reverse transcription: Add Trizol lysate to the colon tissue to fully homogenize, add chloroform to extract to obtain the upper aqueous phase, add an equal volume of isopropanol and mix well; centrifuge to discard the supernatant, add 75% ethanol to wash the RNA precipitate ; After air-drying, DEPC water was added to dissolve; RNA was reverse-transcribed into cDNA according to the instructions of the reverse transcription kit (Thermo, product number: K1621).

[0057] (2) RT-PCR: 10 μl reaction system: 5 μl TB reaction solution, 0.2 μl ROX dye, 0.4 μl each of upstream and downstream primers (10 μM); 1 μl cDNA; 3 μl enzyme-free water.

[0058] Step 1: 95°C, 5min

[0059] Step 2: 95°C, 15s; 60°C, 15s; ...

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Abstract

The invention belongs to the technical field of medicines, and particularly relates to application of insulin-like growth factor 2 recombinant protein in preparation of medicines for treating ulcerative colitis. Experiments show that intraperitoneal injection of the IGF2 recombinant protein can effectively reduce the levels of colitis factors IL1 beta and TNF alpha of ulcerative colitis mice, andrecover the expression levels of insulin-like growth factor type 1 receptors, insulin-like growth factor 2 binding proteins 2 and tight junction proteins ZO1 and Occludin in colons. In addition, the colon length of the DSS model mouse can be obviously recovered through IGF2 recombinant protein administration. Results show that the IGF2 recombinant protein can reduce the inflammatory factor level in colon and improve the expression level of tight junction proteins by up-regulating the expression of IGF1R and IGF2BP2, so that the colon injury of ulcerative colitis mice is effectively prevented,and the IGF2 recombinant protein can be used for preparing a medicinal preparation for treating ulcerative colitis.

Description

technical field [0001] The invention belongs to the technical field of medicine, and in particular relates to the application of insulin-like growth factor 2 recombinant protein in the preparation of medicines for treating ulcerative colitis. Background technique [0002] Ulcerative colitis (UC) is a non-specific inflammatory bowel disease of ulcerative colitis, which is called idiopathic colitis by the World Health Organization. It is a chronic nonspecific inflammatory process of unknown etiology limited to the colonic mucosa and submucosa. The lesions are mostly located in the sigmoid colon and rectum, and can also extend to the descending colon or even the entire colon. Its main feature is that the colorectal mucosa presents continuous diffuse inflammation, and its clinical manifestations are diarrhea, mucus, pus and blood in the stool, abdominal pain, etc., and its specific pathogenesis has not yet been fully elucidated. UC is difficult to cure, long-term persistent, po...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K38/30A61P1/00A61P1/04
CPCA61K38/30A61K9/0019A61P1/00A61P1/04
Inventor 陈玲曹文宇钟小林刘丹丹王雅娟许晓帆
Owner THE FIRST AFFILIATED HOSPITAL HENGYANG MEDICAL SCHOOL UNIV OF SOUTH CHINA
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