Method for investigating mechanism of action of traditional Chinese medicine
A technology of action mechanism and determination method, which is applied in the field of biosensing analysis technology for the determination of the action mechanism of traditional Chinese medicines, can solve the problems that the influence of the combination of effective components of traditional Chinese medicines is not detected, and the patent literature reports on the mechanism of action of traditional Chinese medicines are not found. To avoid the influence of subjective factors and achieve the effect of good repeatability
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Embodiment 1
[0058] The selected traditional Chinese medicine is Radix Paeoniae Rubra, which promotes blood circulation and removes blood stasis, and its active ingredient is Chishao 801; the selected cytokines and their receptors are human recombinant cytokine endothelin-1 (ET-1) and human recombinant cytokine Receptor (ET-1 receptor); The selected activated biotin is SULFO-NHS-LC-BIOTIN; The selected avidin is Strepavidin; The selected biosensing system is the SEIKO QCA917 type of EG&G Company, with Winchem software and The surface area is 0.19±0.01×10 -4 m 2 , Thickness 0.3mm, F 0 =9MHz AT-cut gold quartz crystal substrate piezoelectric quartz crystal microbalance.
[0059] In the first step, add 4-100 μg of human recombinant cytokine endothelin-1 into 200-500 μL of phosphate buffer solution with a concentration of 0.01mol / L and pH=7.4; activate biotin with 1-2 mg of SULFO-NHS-LC-BIOTIN Dissolve in 120 μL of deionized water, take 20-50 μL of it and add it to the above-mentioned human...
Embodiment 2
[0075] Except that the traditional Chinese medicine Chishao 801 used is replaced by Ligustrazine, the active ingredient of Ligusticum Chuanxiong, the others are the same as in Example 1. The test results of this experiment show that Ligustrazine can specifically interact with human recombinant cytokine endothelin-1 and its receptor. Sexual combination, suggesting that the target of ligustrazine in the body is human recombinant cytokine endothelin-1 and its receptor pathway in addition to the reported thromboxane A2. It inhibits the combination of human recombinant cytokine endothelin-1 and human recombinant cytokine endothelin-1 receptor, thereby antagonizing the effect of endothelin, dilating blood vessels and inhibiting platelet aggregation.
Embodiment 3
[0077] Except that the traditional Chinese medicine Radix Paeoniae Rubra 801 is replaced by ferulic acid, the active ingredient of Chinese medicine Angelica sinensis, the rest are the same as in Example 1. The test results of this experiment show that ferulic acid can interact with human recombinant cytokine endothelin-1 and its receptor The specific binding occurs, suggesting that the target of ferulic acid in vivo is not only the reported thromboxane A2, but also human recombinant cytokine endothelin-1 and its receptor pathway, that is, ferulic acid binds to human recombinant cytokine The combination of endothelin-1 and its receptor inhibits the combination of human recombinant cytokine endothelin-1 and human recombinant cytokine endothelin-1 receptor, thereby antagonizing the effect of endothelin, dilating blood vessels and inhibiting platelet aggregation.
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