Medicament for preventing and treating neurologic damage and related diseases thereof

A technology for nerve injury and disease, applied in the field of in vitro culture of adult stem cells, mixture of neurotrophic factors, and preparation of drugs for the prevention and/or treatment of nerve injury or related diseases, which can solve problems such as adverse reactions and poor effects

Inactive Publication Date: 2011-01-12
北京脉迪法莫医药科技有限公司
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Further studies have found that the proliferation and differentiation of endogenous progenitor cells induced by nerve injury can be further stimulated by certain neurotrophic factors, but using any one of these factors alone requires high concentrations and the effect is not good, and often causes Adverse effects, especially with vehicle

Method used

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  • Medicament for preventing and treating neurologic damage and related diseases thereof
  • Medicament for preventing and treating neurologic damage and related diseases thereof
  • Medicament for preventing and treating neurologic damage and related diseases thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0041] First, the obtained adipose tissue was digested with 1 mg / ml type I collagenase at 37° C. for 1 hour. Then add 50ml of DMEM medium to the digestion solution, and centrifuge at 200g for 5 minutes. Remove the supernatant, keeping the cell fraction at the bottom of the centrifuge tube. The cells were resuspended in fresh DMEM medium, and then the cell suspension was filtered through a 100 μm cell sieve. The cell suspension was centrifuged at 200g for 5 minutes. The supernatant was removed, and the lower cells were treated with erythrocyte lysate at 37°C for 5 minutes. Then centrifuge at 300g for 5 minutes. The obtained cells were resuspended in microvascular endothelial cell medium, and then 4 × 10 6 piece / cm 2 The cell concentration was inoculated in uncoated T75 tissue culture flasks in CO 2 cultured in an incubator (37°C, 5% CO 2 ).

Embodiment 2

[0043] ASC secreted substances were tested for their ability to stimulate endogenous neurogenesis in the rat hypoxic-ischemic brain. ASCs isolated from rat subcutaneous adipose tissue were grown to confluence in EGM2MV medium, and then transferred to Eagle minimal medium (containing 5 mM K + BME, Invitrogen) for 24 hours (in CO 2 Cultured in an incubator at 37°C, 5% CO 2). The culture (ASC-CM) was harvested and concentrated 50-fold using CentriPlus (MW cut-off: 10k). Neonatal (7 day old) rats were subjected to unilateral (left) carotid artery ligation and then exposed to hypoxic conditions (7% oxygen) for 2 hours. 10 μl of concentrated (50-fold) ASC-CM was injected intravenously 1 hour before hypoxia-ischemia (H-I). After 7 days, comparisons were made to animals treated with H-I only. The results showed that ASC-CM pretreatment significantly stimulated the formation of hippocampal neurons induced by hypoxic-ischemic brain injury (108±43, n=3 vs. 53±17, n=3; see figure 1 ...

Embodiment 3

[0045] To further evaluate how concentrated ASC-CM stimulates neurogenesis in vivo, we used the PC12 cell model to investigate whether ASC-CM affects neuronal differentiation by observing the effect of ASC-CM on synapse outgrowth in these cells. The data showed that ASC-CM significantly stimulated the neuronal differentiation of PC-12 cells (see figure 2 ), and showed that this effect was significantly blocked by a specific inhibitor of AMPK kinase (see Figure 3), suggesting that AMPK plays an important role in NGF-induced neuronal differentiation. AMPK is activated by a variety of pathological stresses including hypoxia, oxidative stress, glucose deprivation, and exercise and dietary hormones such as leptin and adipocytokines. AMPK activation plays a protective role against pathological stress, especially ischemic-hypoxic stress, and can reduce infarct volume. AMPK is also activated in hypothalamic neurons under conditions of dietary restriction. It was also recently shown...

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Abstract

The invention provides a medicament for preventing and treating neurologic damage and related diseases thereof, and in particular relates to the application of a neurotrophic factor mixture secreted by adult stem cells from fat or an adult stem cell in vitro culture from the fat in preventing and/or treating the neurologic damage and the related diseases thereof. The neurotrophic factor mixture exists in the cell in vitro culture; the neurotrophic factor mixture and the cell in vitro culture can generate a synergistic effect, which reinforces activity greatly, reduces clinical dose obviously, reduces treating cost and reduces side effects; and the medicament can be effectively used for the neurologic damage and the related diseases thereof, such as preventing and/or treating the anoxic ischemic encephalopathy without processing steps such as further extracting, purifying and the like or adding a carrier, which fills the gap of the deficiency of conventional preventing and treating means for the neurologic damage and the related diseases thereof, simultaneously reduces the side effects and untoward effects, and further contributes to the reduction of the production cost of the medicament and popularization and application.

Description

technical field [0001] The present invention relates to a medicine for preventing and / or treating nerve injury or its related diseases, in particular to a mixture of neurotrophic factors secreted by adipose-derived adult stem cells or in vitro culture of adipose-derived adult stem cells in the preparation of prevention and / or treatment Use in medicine for nerve damage or related diseases. Background technique [0002] Adipose-derived adult stem cells (ASCs) are mesenchymal stem cells extracted from adipose tissue, which provide an abundant, readily available and replenishable raw material for potential clinical applications. These pluripotent cells are found in the "stromal" or "non-adipocyte" fraction of adipose tissue, previously thought to be "preadipocytes." Because of their ability to differentiate into adipocytes, ASCs have been studied for decades as potential soft tissue substitutes (Dardick, I., et al., Ultrastructural observations on differentiating human preadipo...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K35/12A61K38/18A61K38/30C12N5/0775A61P7/00A61P9/06A61P9/10A61P25/00A61P25/08A61P25/14A61P25/16A61P25/28A61K35/28
Inventor 杜燕生
Owner 北京脉迪法莫医药科技有限公司
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