Protection and action mechanisms of combination of gastrodin and rhynchophylla to vascular endothelial cells

A technique for endothelial cells and rhynchophylline, which is applied in the field of cell biology and can solve the problems of complex composition, synthesis limitation, eNOS inactivation, etc.

Inactive Publication Date: 2013-12-18
CHINA PHARM UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, the development of traditional Chinese medicine has been plagued by problems such as complex ingredients and unclear mechanism of action of traditional Chinese medicine for a long time.
When eNOS binds to the CSD, eNOS is inactivated, resulting in restricted NO synthesis

Method used

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  • Protection and action mechanisms of combination of gastrodin and rhynchophylla to vascular endothelial cells
  • Protection and action mechanisms of combination of gastrodin and rhynchophylla to vascular endothelial cells
  • Protection and action mechanisms of combination of gastrodin and rhynchophylla to vascular endothelial cells

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0027] Embodiment 1 The effect of gastrodin rhynchophylline compatibility on the viability of ECV-304 cells

[0028] Grouping: (1) control group: add 10% DMEM medium; (2) model group: 1000μmol L -1Hcy; (3) Gastrodin group: 100 μg·mL -1 (4) Rhynchophylline group: 100 μg·mL -1 (5) Experimental group: add 10% DMEM medium containing different concentrations of gastrodin rhynchophylline (0.1-10):1 compatibility; (6) Zeroing group: add cell-free medium. Each group has 6 replicate wells.

[0029] Detection of cell viability: (1) EVC-304 cells in logarithmic phase were collected, and after the cells were 80-90% full, 100 μL of drug solutions of different concentrations were added to the experimental group. Add 100 μL of culture medium to the control group and the zeroing group. The final volume of each well is 100 μL. (2) 5%CO 2 , respectively incubated at 37°C for 12, 24, 36, and 48 hours, and observed under an inverted microscope. 16 μL of MTT solution was added to each well,...

Embodiment 2

[0031] Example 2 Determination of the protective effect of gastrodin and rhynchophylline compatibility on the ECV-304 cell model of HCY damage

[0032] According to the drug concentration and grouping determined in Example 1, 4 replicate wells were set up for each group. Measure various biochemical indexes according to lactate dehydrogenase (LDH) kit, malondialdehyde (MDA) kit, NO kit instruction method, test result is as follows Figure 2 ~ Figure 4 shown. According to the results, compared with the model group, the compatibility of gastrodin and rhynchophylline can reduce the release of MDA and LDH, and can increase the release of NO in vascular endothelial cells, and the protection effect on ECV-304 cells is more significant than that of the single component with the same concentration. Example 3 Effect of gastrodin and rhynchophylline compatibility on the expression of CAV-1 protein in ECV-304 cells

Embodiment 3

[0033] The total protein in the cells was extracted, and the target band was analyzed by the Western Blot detection gel image processing system. Analysis results such as Figure 5 , Figure 6 shown. Compared with the control group, the model group can increase the expression of CAV-1 in cells. In the experimental group, low concentration of drugs (5 μg·mL -1 and 10μg·mL -1 ) can inhibit the expression of CAV-1 in Hcy-damaged cells, and high-concentration drugs (50 μg·mL -1 and 100μg·mL -1 ) had no significant inhibitory effect on the expression of CAV-1 in injured cells. The results indicated that the combination of gastrodin and rhynchophylline has the characteristics of irreversible recovery for the protection of Hcy-induced vascular endothelial cell injury, and the mechanism of action is through the CAV-1 / eNOS / NO signaling pathway, so as to achieve the effect of lowering blood pressure.

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Abstract

The invention discloses a protective effect of a combination of gastrodin and rhynchophylla to human vein vascular endothelial cells. Through adding homocysteine with a certain concentration to induce and damage the endothelial cells, a hypertension cell damage model is built. By using the model, combination drugs of gastrodin and rhynchophylla with various concentrations are added in the model, the biochemical indexes such as lactic dehydrogenase (LDH), malondialdehyde (MDA) and nitric oxide (NO) are detected, so as to test the protective effect of the combination drug to the damaged endothelial cells. The expression of key protein on a latent signal channel is observed by the Western Blotting technology, the observation proves that the latent channel is the cell signal channel for the combination drug to generate the protective effect. In the invention, the effective constituents in two natural blood-pressure reducing drugs of gastrodin and rhynchophylla are creatively combined, and the protective effect of the combination drug to the hypertension cell damage model is researched. The effect of the combination is more remarkable than that of any single component, and the mechanism is clear, therefore the combination drug can become the antihypertensive drug of secondary development of the traditional Chinese medicine.

Description

technical field [0001] The invention belongs to the technical field of cell biology, and relates to the compatibility of gastrodin and rhynchophylline, two natural antihypertensive drug active ingredients of Gastrodia elata and Uncaria, in particular to the protection and action mechanism of the compatibility of the two ingredients on human vein endothelial cells. Background technique [0002] Gastrodia elata is a precious traditional Chinese medicine. It is often used clinically for headaches, dizziness, numbness of limbs, convulsions in children, epilepsy, convulsions, and tetanus. Gastrodia elata contains a variety of chemical components, such as gastrodin, gastrodin glycosides, gastrodia elata polysaccharide, vanilla alcohol, vanilla aldehyde, p-hydroxybenzyl alcohol, citric acid, succinic acid, etc. [0003] Gastrodin is the most abundant single component that can be isolated so far, and its structure is p-hydroxymethylbenzene-β-D-glucopyranoside. At present, gastrodin...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A61K31/7034A61P9/12A61K31/438
Inventor 许激扬卞筱泓沃龙飞危喜玲
Owner CHINA PHARM UNIV
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