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CN118987175A - Application of plexafet and granulocyte colony stimulating factor in preparation of medicine for treating cerebral infarction - Google Patents

Application of plexafet and granulocyte colony stimulating factor in preparation of medicine for treating cerebral infarction Download PDF

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Publication number
CN118987175A
CN118987175A CN202411105270.9A CN202411105270A CN118987175A CN 118987175 A CN118987175 A CN 118987175A CN 202411105270 A CN202411105270 A CN 202411105270A CN 118987175 A CN118987175 A CN 118987175A
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Prior art keywords
cerebral infarction
stimulating factor
granulocyte colony
colony stimulating
rats
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Inventor
杨业国
苏晓萍
侯海燕
王仕米
邓佐成
刘宇翔
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Guangdong Yijia Regenerative Medicine Research Institute Co ltd
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Guangdong Yijia Regenerative Medicine Research Institute Co ltd
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    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K38/00Medicinal preparations containing peptides
    • A61K38/16Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • A61K38/17Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • A61K38/19Cytokines; Lymphokines; Interferons
    • A61K38/193Colony stimulating factors [CSF]
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61KPREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00Medicinal preparations containing organic active ingredients
    • A61K31/33Heterocyclic compounds
    • A61K31/395Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • AHUMAN NECESSITIES
    • A61MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61PSPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P9/00Drugs for disorders of the cardiovascular system
    • A61P9/10Drugs for disorders of the cardiovascular system for treating ischaemic or atherosclerotic diseases, e.g. antianginal drugs, coronary vasodilators, drugs for myocardial infarction, retinopathy, cerebrovascula insufficiency, renal arteriosclerosis

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  • Chemical & Material Sciences (AREA)
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  • Animal Behavior & Ethology (AREA)
  • Medicinal Chemistry (AREA)
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  • Heart & Thoracic Surgery (AREA)
  • Organic Chemistry (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • General Chemical & Material Sciences (AREA)
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  • Urology & Nephrology (AREA)
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  • Proteomics, Peptides & Aminoacids (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)

Abstract

The invention provides an application of pleoxate and granulocyte colony stimulating factor in preparing a medicine for treating cerebral infarction, and belongs to the technical field of biological medicines. The cerebral infarction is acute cerebral infarction. According to the invention, through a rat experiment, the effect of increasing the weight of a rat, reducing the nerve function score and the volume percentage of cerebral infarction by combining the Prlay and the granulocyte colony stimulating factor administration is achieved, and compared with the effect of reducing the cerebral infarction degree, reducing the volume of cerebral infarction and promoting the recovery of nerve function by singly administering the granulocyte colony stimulating factor, the Prlay and the granulocyte colony stimulating factor administration have better effect of reducing the cerebral infarction degree of the rat.

Description

Application of plexafet and granulocyte colony stimulating factor in preparation of medicine for treating cerebral infarction
Technical Field
The invention relates to the technical field of biological medicines, in particular to application of pleoxate and granulocyte colony stimulating factor in preparation of a medicine for treating cerebral infarction.
Background
Cerebral infarction, also called ischemic cerebral apoplexy, is a cerebrovascular disease caused by insufficient cerebral blood supply and oxygen supply due to the blockage of cerebral blood flow supply, and the main morbidity group is middle-aged and elderly people, and has the advantages of urgent morbidity, higher disability rate and death rate, and poorer prognosis effect. Treatment of cerebral infarction is limited due to non-reproducibility of neurons after necrosis and abnormal establishment of neurite function. At present, the clinical treatment of cerebral infarction mainly adopts drug conservation treatment, such as a thrombus dredging capsule, but after the drug treatment of partial patients, symptoms are not obviously improved, the risk of the nerve function injury of the patients is easily increased, and the cerebral infarction has certain limitation.
Granulocyte colony stimulating factor (granulocyte colony-stimulating factor, G-CSF), a glycoprotein produced by vascular endothelial cells, monocytes and fibroblasts, binds to specific receptors on the cell surface, promotes neutrophil maturation, stimulates release of mature granulocytes from bone marrow, and enhances neutrophil chemotaxis and phagocytosis. Studies have shown that granulocyte colony-stimulating factor can treat acute cerebral infarction in rats by mobilizing bone marrow stem cells, but clinical studies have shown that granulocyte colony-stimulating factor does not improve the clinical outcome in patients with acute cerebral infarction. Therefore, it is necessary to provide a new medicine for treating cerebral infarction, so as to provide a new idea for clinical treatment of cerebral infarction.
Disclosure of Invention
The invention aims to provide an application of plexacum in preparing a medicament for treating cerebral infarction by combining with granulocyte colony stimulating factor, and provides a new thought for clinical treatment of cerebral infarction.
In order to achieve the above object, the present invention provides the following technical solutions:
The invention provides an application of pleoxacin and granulocyte colony stimulating factor in preparing a medicine for treating cerebral infarction.
Preferably, the cerebral infarction is an acute cerebral infarction.
The invention adopts a wire bolt method to prepare a rat cerebral infarction model, and randomly divides rats with successful modeling and nerve function scores of 2-3 into an experimental group, a control group and a model control group. The experimental group was intraperitoneally injected with granulocyte colony-stimulating factor (G-CSF) at 8 a.m., 30. Mu.g/kg; at 20 night, common Le Sha mg/kg was injected intraperitoneally. Rats in the control group were intraperitoneally injected with G-CSF at 30 μg/kg at 8 a.m. per day; at night 20, equal amount of physiological saline was injected intraperitoneally. Model control rats were injected with an equal amount of physiological saline at 8 a.m. and 20 a.m. each day. According to the invention, through a rat experiment, the effect of increasing the weight of a rat, reducing the nerve function score and the volume percentage of cerebral infarction by combining the Prlay and the granulocyte colony stimulating factor administration is achieved, and compared with the effect of reducing the cerebral infarction degree, reducing the volume of cerebral infarction and promoting the recovery of nerve function by singly administering the granulocyte colony stimulating factor, the Prlay and the granulocyte colony stimulating factor administration have better effect of reducing the cerebral infarction degree of the rat.
Detailed Description
The technical solutions provided by the present invention are described in detail below with reference to examples, but they should not be construed as limiting the scope of the present invention.
Example 1
SPF-grade SD male rats with weights between 250 and 300g are selected, and are adaptively fed for 7 days under the conditions of 25 ℃ and 55% relative humidity and 12h/12h of light and dark period, and the rats are free to drink and ingest during feeding. After the adaptive feeding is finished, a rat cerebral infarction model is prepared by adopting a wire bolt method, and the specific steps are as follows:
Rats were anesthetized by intraperitoneal injection of 10% chloral hydrate (0.5 mL/100 g) and the surgical site was dehaired and aseptically treated, the median skin beside the left neck of the rat was incised, and the left common carotid artery was isolated to the carotid artery and carotid artery bifurcation. Separating and ligating the proximal end of the external carotid artery along the bifurcation of the internal carotid artery, clamping the proximal end and the distal end of the common carotid artery by an arterial clamp, tying a loose knot at the proximal end of the common carotid artery, cutting a small opening at a position about 4mm away from the ligature position, inserting a nylon wire with the diameter of 0.24mm and a fire-burned tip, loosening the arterial clamp at the distal end of the common carotid artery, and lightly pushing the tail end of the nylon wire to enable the tail end of the nylon wire to enter the internal carotid artery from the common carotid artery through the bifurcation, wherein the insertion depth is 18mm. And (3) fastening the loose knot at the proximal end of the common carotid artery, ligating the distal end of the common carotid artery, removing an arterial clamp, cutting off redundant nylon wires, suturing a wound and carrying out single-cage feeding.
The same side Horner syndrome of the operation appears after the rat wakes up, the right forelimb is bent and adduction appears on the opposite side of the operation and can not be fully extended, and the moulding is successful after the rat is placed on the ground to form a circle on the right side and rear-end collision. After modeling is successful for 24 hours, neural function scoring is carried out, and scoring standards are as follows: 0 point: no symptom of nerve injury; 1, the method comprises the following steps: the contralateral forelimb cannot be fully extended; 2, the method comprises the following steps: the walking is carried out to the contralateral rotating ring; 3, the method comprises the following steps: unstable standing and toppling to the opposite side; 4, the following steps: can not walk independently, and the consciousness is lost. Rats with scores of 2-3 are selected as test animals, 36 rats are selected and randomly divided into 3 groups, namely an experimental group, a control group and a model control group, and each group comprises 12 rats.
The rats of the experimental group were intraperitoneally injected with granulocyte colony-stimulating factor (G-CSF) at 8 a.m. per day; at 20 night, common Le Sha mg/kg was injected intraperitoneally. Rats in the control group were intraperitoneally injected with G-CSF at 30 μg/kg at 8 a.m. per day; at night 20, equal amount of physiological saline was injected intraperitoneally. Model control rats were injected with an equal amount of physiological saline at 8 a.m. and 20 a.m. each day. The test was performed for a total of 7 days. After the test is finished, the rats in each group are continuously fed at the temperature of 25 ℃ and the relative humidity of 55 percent under the condition of 12 hours/12 hours of light and shade period, and the rats are free to drink water and eat during the feeding period.
Example 2
1. Weight measurement
The body weights of the rats of each treatment group in example 1 were measured before the test and on the 7 th and 21 st days after the end of the test, and the measurement results are shown in table 1.
Table 1 results of weight measurements before and after the treatment group rats were tested
As can be seen from table 1, there was no significant difference in body weight between the rats in the treatment groups prior to the test; on day 7 after the test, the rats in each treatment group had different degrees of weight loss due to reduced mobility and reduced feeding, and there was no obvious difference between the weights of the rats in the experimental group and the control group, but it was significantly higher than that in the model control group; on day 21 after the test, the body weight of the rats in each treatment group was increased compared to day 7 after the test, and the body weight of the rats in the test group was higher than that of the control group and significantly higher than that of the model control group due to the recovery of the mobility of the rats. It can be seen that the administration of plexacin in combination with granulocyte colony stimulating factor is superior to the administration of granulocyte colony stimulating factor alone for the degree of weight gain in rats.
2. Neural function score determination
The neurological scores were measured on days 7 and 21, respectively, after the end of the treatment group rats in example 1, and the measurement results are shown in table 2.
Table 2 results of determination of neurological score after rat experiments in each treatment group
Day 7 after the test Day 21 post-test
Experimental group 2.67 0.83
Control group 2.42 1.25
Model control group 2.75 2.33
As can be seen from table 2, the difference in the nerve function score of rats in each treatment group at 7 days after the test is not obvious, but the nerve function score of rats in the group to which the plexacin is administered with the combined granulocyte colony stimulating factor is obviously reduced with the increase of time, which indicates that the administration of the plexacin with the combined granulocyte colony stimulating factor has a better improvement effect on cerebral infarction of rats.
3. Volume percent determination of cerebral infarction
On day 21 after the end of the test, 6 rats were randomly selected from each treatment group, anesthetized with 10% chloral hydrate (0.5 mL/100 g) by volume fraction, craniocated, and then placed in a petri dish and rapidly placed in a-20℃refrigerator for 20min for cold storage and then removed. Slicing brain tissue along coronal plane at intervals of 2mm, soaking the sliced brain tissue slices in 2,3, 5-triphenyltetrazolium chloride dye solution, wrapping with tinfoil paper, and placing in a 37 ℃ incubator for dyeing for 30min, and continuously turning the brain tissue slices to uniformly dye. Photographing is carried out after dyeing is completed, analysis is carried out by using Image J software, the volume percent of cerebral infarction is measured, and the measurement results are shown in table 3.
Percent infarct volume = infarct volume/total brain volume x 100%.
Table 3 results of measurement of volume percent cerebral infarction in rats of each treatment group
Volume percent cerebral infarction (%)
Experimental group 12.39
Control group 16.58
Model control group 23.64
As can be seen from table 3, compared with the control group and the model control group, the experimental group can significantly reduce the volume percentage of cerebral infarction in rats, and the administration of the plexacin combined with the granulocyte colony stimulating factor has better effect of improving cerebral infarction compared with the administration of the granulocyte colony stimulating factor alone.
From the above examples, the present invention provides the use of plexacum in combination with granulocyte colony stimulating factor in the preparation of a medicament for treating cerebral infarction. According to the invention, through a rat experiment, the effect of increasing the weight of a rat, reducing the nerve function score and the volume percentage of cerebral infarction by combining the Prlay and the granulocyte colony stimulating factor administration is achieved, and compared with the effect of reducing the cerebral infarction degree, reducing the volume of cerebral infarction and promoting the recovery of nerve function by singly administering the granulocyte colony stimulating factor, the Prlay and the granulocyte colony stimulating factor administration have better effect of reducing the cerebral infarction degree of the rat. The administration of the plexafex-associated granulocyte colony stimulating factor is expected to become a novel method for clinical treatment of cerebral infarction.
The foregoing is merely a preferred embodiment of the present invention and it should be noted that modifications and adaptations to those skilled in the art may be made without departing from the principles of the present invention, which are intended to be comprehended within the scope of the present invention.

Claims (2)

1. Application of plexafex-combined granulocyte colony stimulating factor in preparing medicine for treating cerebral infarction.
2. The use according to claim 1, wherein the cerebral infarction is an acute cerebral infarction.
CN202411105270.9A 2024-08-13 2024-08-13 Application of plexafet and granulocyte colony stimulating factor in preparation of medicine for treating cerebral infarction Pending CN118987175A (en)

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Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101389329A (en) * 2006-02-24 2009-03-18 健赞股份有限公司 Methods of increasing blood flow and/or promoting tissue regeneration
CN102302493A (en) * 2001-07-31 2012-01-04 阿诺麦德股份有限公司 Methods to mobilize progenitor/stem cells
US20120225028A1 (en) * 2011-03-02 2012-09-06 Moshe Cohen Compositions and methods for mobilization of stem cells
KR20140019492A (en) * 2012-08-03 2014-02-17 가톨릭대학교 산학협력단 A method for mobilization of cd11b+cx3cr1+ cells
US20140219952A1 (en) * 2010-03-23 2014-08-07 The Johns Hopkins University Methods of treatment using stem cell mobilizers

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102302493A (en) * 2001-07-31 2012-01-04 阿诺麦德股份有限公司 Methods to mobilize progenitor/stem cells
CN101389329A (en) * 2006-02-24 2009-03-18 健赞股份有限公司 Methods of increasing blood flow and/or promoting tissue regeneration
US20140219952A1 (en) * 2010-03-23 2014-08-07 The Johns Hopkins University Methods of treatment using stem cell mobilizers
US20120225028A1 (en) * 2011-03-02 2012-09-06 Moshe Cohen Compositions and methods for mobilization of stem cells
KR20140019492A (en) * 2012-08-03 2014-02-17 가톨릭대학교 산학협력단 A method for mobilization of cd11b+cx3cr1+ cells

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
LIPING ZHOU等: "Granulocyte-colony stimulating factor in combination with AMD3100 confers greater neuroprotection after hypoxic-ischemic brain damage than a solitary treatment in mice.", 《INT J CLIN EXP MED》, vol. 11, no. 10, 30 October 2018 (2018-10-30) *
郭慧娟等: "小鼠脑梗死后骨髓极小胚胎样干细胞的动员", 《基础医学与临床》, vol. 31, no. 11, 30 November 2011 (2011-11-30), pages 1195 *

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