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A pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application

A composition and drug technology, applied in the field of pharmaceutical compositions against cerebral ischemia-reperfusion injury, can solve the problems of unstable quality, complex and complex components, and difficult to clarify the mechanism of efficacy, so as to restore brain function and reduce cell apoptosis. , Enhanced effect of anti-cerebral ischemia-reperfusion injury

Active Publication Date: 2021-07-09
GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The technical problem to be solved by the present invention is to overcome the technical problems that the existing Buyang Huanwu Decoction traditional Chinese medicine prescription compatibility still stays at the compatibility of traditional Chinese medicine decoction pieces, the ingredients are complex and complicated, the quality is unstable, and the curative effect mechanism is difficult to be clear. The pharmaceutical composition against cerebral ischemia-reperfusion injury, through the experimental research on the synergistic effect of the active components of Buyang Huanwu Decoction on ischemic stroke, has certain safety and efficacy, and has specific functions Combination of effective components, so as to ensure accurate clinical dosage and safe and effective, which is conducive to the development of modern new Chinese medicine with clear ingredients and clear mechanism

Method used

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  • A pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application
  • A pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application
  • A pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Example 1 Isolation, cultivation and identification of primary rat brain microvascular endothelial cells (rBMECs)

[0034] 1. Preparation of reagents

[0035] (1) 20% serum complete medium: Add 5mL penicillin-streptomycin solution and 100mL fetal bovine serum to 400mL DMEM / F12 medium, shake well, aliquot into 50mL centrifuge tubes, and store in a 4°C refrigerator for later use.

[0036] (2) 0.1% type II collagenase solution: add 20mL DMEM / F12 medium to 100mg type II collagenase powder to make a 0.5% type II collagenase mother solution, pass through a 0.22μm filter membrane to filter and sterilize, and absorb 5mL , add 20mL DMEM / F12 medium, mix well to obtain 0.1% type II collagenase solution, dispense it into 15mL, and store it in a -20°C refrigerator for later use.

[0037] (3) 25% bovine serum albumin (BSA) solution: transfer 10g of BSA powder to a 50mL centrifuge tube, add 40mL of DMEM / F12 basic medium, shake to dissolve slowly, pass through a 0.22μm microporous mem...

Embodiment 2

[0059] Example 2 Effects of three active ingredients on the activity of oxygen-glucose deprived brain microvascular endothelial cells

[0060] 1. Preparation of a series of dosing solutions

[0061] Weigh 2mg standard substances of verbascoside, formononetin, and 7-2'-dihydroxy-3',4'-dimethoxyisoflavane respectively, add them to 0.2mL EP tube, and add 50μL DMSO to carry out Dissolved, prepared as 40mg / mL mother solution, and diluted with complete medium to 0.1, 0.5, 1, 5, 10, 20, 40 μg / mL serial administration solutions (DMSO concentration controlled below 0.1%).

[0062] 2. Preparation of VEGF solution

[0063] Take 10μg VEGF and dissolve it in 10mL PBS solution, the concentration is 1μg / mL at this time, and distribute it for use. Pipette 10 μL of the above solution into a sterilized EP tube, and then add 990 μL of complete medium, and the concentration at this time is 10 ng / μL.

[0064] 3. Cell inoculation

[0065] Cells at 25cm 2 When cultured in the culture flask to 8...

Embodiment 3

[0083] Example 3 Effects of Active Ingredient Combination on the Expression of Cerebral Microvascular Endothelial Cells VEGF, bFGF, LDH and SOD

[0084] 1. Extraction and separation of primary rBMECs

[0085] Refer to the extraction and separation of primary rBMECs in Step 2 of Example 1.

[0086] 2. Subculture of cells

[0087] Referring to Step 3 of Example 1, the cells were subcultured.

[0088] 3. Cell inoculation

[0089] Refer to Step 3 of Example 2 for cell inoculation.

[0090] 4. Establishment of hypoxic injury model of rat cerebrovascular endothelial cells

[0091] Referring to step 4 of Example 2, the experimental grouping (2) construction of the model group.

[0092] 5. Grouping

[0093] According to the results of the CCK-8 cytotoxicity test, the concentrations of calycosin, formononetin, and 7-2'-dihydroxy-3',4'-dimethoxyisoflavane were in the range of 1-20, 0.1-10, 1-20 μg / mL is a non-cytotoxic dose, that is, after 24 hours of co-culture of each drug and ...

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Abstract

The invention discloses a pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application. The pharmaceutical composition is composed of the following components at final concentrations: 1-10 μg / mL verbascoside, 0.1-1 μg / mL formononetin, 0.5-5 μg / mL7-2'-dihydroxy-3',4 '‑Dimethoxyisoflavane. In the present invention, by optimizing the proportion of the three effective active ingredients in Buyang Huanwu Decoction, the anti-cerebral ischemia-reperfusion injury effect of the pharmaceutical composition is significantly enhanced, and the cell apoptosis caused by cerebral ischemia-reperfusion injury can be alleviated. , improve the survival rate of brain microvascular endothelial cells, and can increase the expression levels of vascular endothelial growth factor, fibroblast growth factor, lactate dehydrogenase and superoxide dismutase in brain microvascular endothelial cells, and help restore brain function, It can be used to prepare medicines or preparations for preventing or treating ischemic cerebral apoplexy, and has broad application prospects.

Description

technical field [0001] The invention relates to the technical field of traditional Chinese medicine, in particular to a pharmaceutical composition for resisting cerebral ischemia-reperfusion injury and its application. Background technique [0002] Ischemic stroke (ischemic stroke) is one of the most common cerebrovascular diseases. It has the characteristics of high morbidity, high disability rate, high mortality rate, high recurrence rate and many complications. It is imperative to reduce the incidence, disability and mortality of stroke. Studying the pathogenesis of cerebral apoplexy and its prevention and treatment measures has not only extremely important social and practical significance, but also has high theoretical value and clinical practical significance. [0003] The active ingredients of traditional Chinese medicine are the material basis for the prevention and treatment of diseases in traditional Chinese medicine. How to make the active ingredients produce the...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A61K31/7048A61K31/353A61P9/10
CPCA61K31/353A61K31/7048A61P9/10A61K2300/00
Inventor 王利胜沈晓张艳张英丰廖丰蕴
Owner GUANGZHOU UNIVERSITY OF CHINESE MEDICINE
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