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Application of CXCL16 protein and its monoclonal antibody in preparation of drugs to prevent and/or treat intestinal injuries

A monoclonal antibody and intestinal injury technology, applied in the field of biomedicine, can solve the problem of no report on the effect of CXCL16 in radiation enteritis, and achieve the effects of difficult clinical diagnosis, low cost, and good patient compliance.

Active Publication Date: 2019-09-20
THE SIXTH AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

It has been reported that CXCL16 plays an important role in the occurrence and development of inflammatory bowel disease, but the role of CXCL16 in radiation enteritis has not been reported in the prior art

Method used

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  • Application of CXCL16 protein and its monoclonal antibody in preparation of drugs to prevent and/or treat intestinal injuries
  • Application of CXCL16 protein and its monoclonal antibody in preparation of drugs to prevent and/or treat intestinal injuries
  • Application of CXCL16 protein and its monoclonal antibody in preparation of drugs to prevent and/or treat intestinal injuries

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0022] Example 1 Expression of CXCL16 in radiation enteritis cases

[0023] test subject:

[0024] In order to detect the expression of CXCL16 in radiation enteritis cases, the inventor collected 30 cases of rectal cancer patients with severe radiation enteritis diagnosed by clinical imaging after radiotherapy and 33 cases of mild radiation enteritis diagnosed by imaging from the Sixth Affiliated Hospital of Sun Yat-sen University. Surgical resection of paracancerous intestinal tissue samples and serum samples after radiotherapy. The expression of CXCL16 was detected by ELISA and qRT-PCR respectively.

[0025] experimental method:

[0026] 1. Detect the expression of CXCL16 protein in the serum of samples

[0027] ELISA detection of CXCL16 expression in serum Experimental method and steps: CXCL16 ELISA kit was purchased from Guangzhou Ruiboao Biotechnology Co., Ltd. After the ELISA plate coated with the antibody was equilibrated to room temperature, 100 μL of the prepared s...

Embodiment 2

[0040] Example 2 Expression of CXCL16 in an animal model of radiation enteritis

[0041] 1. Construction of animal model of radiation enteritis

[0042] C57BL / 6J mice were purchased from the Institute of Model Animals, Nanjing University. When the mice were 8 weeks old, a single irradiation (25Gy) was performed with an X-ray linear accelerator. The mice were fixed after anesthesia, and the exposure range was from the pubic symphysis to the anus (a rectangular area with a length of 1.5 cm and a width of 1.0 cm from the anus). 5 irradiated mice and 5 non-irradiated normal control mice were sacrificed at each time point of 4 hours, 1 day, 3 days, 2 weeks, 8 weeks and 6 months after irradiation. The mouse rectal tissue was collected and stored in RNA later solution.

[0043] 2. Detection of CXCL16 protein and mRNA expression levels in animal model serum and intestinal tissue

[0044] (1) ELISA detection of mouse serum CXCL16 protein expression kit purchase and experimental met...

Embodiment 3

[0059] Example 3 CXCL16 Gene Knockout Significantly Inhibits Radiation Enteritis Fibrosis

[0060] 1. Construction of CXCL16 gene knockout mice

[0061] CXCL16 gene whole-body knockout mice were commissioned to construct by Shanghai Nanfang Model Organisms Co., Ltd., using CRISPR / Cas9 technology, using non-homologous recombination to repair and introduce mutations, resulting in a frame shift of the CXCL16 gene protein reading frame and loss of function. The brief process is as follows: Cas9 mRNA and gRNA were obtained by in vitro transcription; Cas9 mRNA and gRNA were microinjected into fertilized eggs of C57BL / 6J mice to obtain F0 generation mice. F0 generation mice were mated with C57BL / 6J mice to obtain F1 heterozygous mice with CXCL16 gene knockout.

[0062] 2. Establishment of radiation enteritis fibrosis model in CXCL16 knockout mice

[0063] 10 wild-type (WT) and 10 homozygous knockout mice (KO) were obtained by selfing the obtained CXCL16 gene knockout F1 heterozygou...

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Abstract

The invention provides application of CXCL16 protein and its monoclonal antibody in the preparation of drugs to prevent and / or treat intestinal injuries, and also provides a molecular marker to predict or / and diagnose severity of radiation enteritis. The molecular marker is CXCL16 protein, having an amino acid sequence shown as SEQ NO. 1. It is discovered through a series of human sample and animal experimental verifications that a monoclonal antibody of CXCL16 can prevent and relieve radiation intestinal injury and fibrosis effectively and has a good clinical application prospect. The severity of radiation enteritis in a patient can be judged herein by detecting expression level of CXCL15 mRNA in peripheral serum of the patient; judging is simple and fast; patient compliance is good, and the cost is low.

Description

technical field [0001] The invention belongs to the technical field of biomedicine, and relates to a molecular marker of radiation enteritis and the application of a monoclonal antibody thereof, in particular to chemokine CXCL16 as a molecular marker of radiation enteritis in the preparation of a drug for preventing and / or treating intestinal injury diseases. application in medicine. Background technique [0002] Radiation therapy is one of the most effective treatments for pelvic and abdominal malignant tumors, including cervical cancer, ovarian cancer, prostate cancer, rectal cancer and bladder cancer, and can significantly prolong the survival time of patients with advanced tumors. However, radiation damage to normal organs and tissues in the pelvic and abdominal cavity caused by radiotherapy will cause a variety of serious complications and greatly affect the quality of life of patients. Among them, radiation enteritis (Radiation enteritis, RE) is one of the most common...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K39/395A61P1/00C12Q1/6883
CPCC07K16/24A61P1/00C12Q1/6883A61K2039/505C12Q2600/158C12Q2600/166Y02A50/30
Inventor 王磊崔艳梅
Owner THE SIXTH AFFILIATED HOSPITAL OF SUN YAT SEN UNIV
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