Pharmaceutical composition providing inhibition effect against inflammation factor expression
A technology of inhibition and inflammatory factors, applied in the field of medicine, can solve the problems of large dosage of original prescription and difficulty of new drug development, etc.
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experiment example 1
[0026] Experimental Example 1 Inhibitory Effect of a Single Extract on the Binding of NF-κB Response Elements
[0027] 1. Experimental steps
[0028](1) Experimental materials: NF-κB-RE cells (pGL4.32[luc2P / NF-kappaB-RE]293A, Institute of Materia Medica, Chinese Academy of Medical Sciences), MEM-α medium (Gibco), fetal bovine serum (Gibco) , 96-well culture plate (Costar, 3599), Bright-Glo Luciferase detection reagent (Promaga), water-jacketed CO 2 Cell incubator (Sanyo, Japan), Safire2 high-speed multi-channel continuous wavelength microplate reader (TECAN, Austria).
[0029] (2) Test samples:
[0030] Extracts 1-3 obtained by the method described in Example 1 were provided by Beijing University of Traditional Chinese Medicine.
[0031] (3) Experimental steps:
[0032] Digest and count the logarithmically grown NF-κB-RE cells in the culture flask, and use 100 μl system, 5×10 3 / well density inoculated in 96-well white culture plate, incubated for 18-24h. The control gro...
experiment example 2
[0041] Experimental Example 2 The agonistic effect of a single extract on peroxisome proliferator receptor (PPARγ)
[0042] 1 material
[0043] 1.1 Reagents:
[0044] African green ape kidney cells (CV-1), PPRE plasmid, PPAR plasmid, RXRα plasmid, MEM-α medium (Gibco), fetal bovine serum (Gibco), Bright-Glo Luciferase luciferase detection reagent (Promega) , rosiglitazone.
[0045] 1.2 Instruments:
[0046] Water-jacketed CO2 cell incubator (Sanyo, Japan), EnSpireTM 2300MultilabelReader (PerkinElmer).
[0047] 2 methods
[0048] 2.1 Extraction of the test product
[0049] Three extracts obtained by the method described in Example 1 were provided by Beijing University of Traditional Chinese Medicine.
[0050] 2.2 Cell Culture
[0051] African green ape kidney cells (CV-1) were cultured in DMEM medium supplemented with 10% FBS and 1% antibiotics (penicillin / streptomycin) in 5% CO 2 Cultured in an incubator at 37°C.
[0052] 2.3 Transfection and luciferase gene expressio...
experiment example 3
[0063] Experimental Example 3 Study on the Combination and Inhibitory Effect of Extracts on NF-κB Response Elements
[0064] 1. Experimental steps
[0065] Same as Experiment 1.
[0066] 2. Orthogonal design
[0067] The research on the compatibility law adopts 3 factors and 3 levels of orthogonal experimental design, according to the orthogonal table, a total of 27 compatibility groups are designed. Among them, the three factors are the first extract, the second extract, and the third extract, and the concentration points of the three levels are taken within the range of the active dose-effect relationship of each active site. The factor levels are shown in Table 3.
[0068] table 3
[0069] (μg / ml) 1st extract 2nd extract 3rd extract 1 69 54 51.6 2 34.5 27 25.8 3 17.25 13.5 12.9
[0070] 3. Experimental results
[0071] See Table 4 for detailed results.
[0072] Table 4 The effect of each compatibility gr...
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