Use of isochlorogenic acid compound and different combinations in hepatitis treatment
A technology of isochlorogenic acid and compounds, applied in the application field of isochlorogenic acid compounds and different combinations in the treatment of hepatitis, to achieve the effect of strong anti-hepatitis B virus, inhibition of replication, and simple and easy extraction process
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Embodiment 1
[0038] The isochlorogenic acid composition can secrete HBsAg (hepatitis B virus surface antigen), HBeAg (hepatitis B virus E antigen) and HBV-DNA (hepatitis B virus) to transfect HBV (hepatitis B virus) gene HepG2. Inhibition of the viral replication number of the virus).
[0039] Determination of the inhibitory effect of the sample on the growth of HepG2.2.15 cells: the cells were digested with trypsin-EDTA and diluted to 1×10 5 / mL, added to a 96-well cell culture plate, 100uL per well, placed in CO 2 cultured in an incubator. After 24 hours of inoculation, discard the medium, add the sample diluted with the medium, 200ul per well, add 3 wells for each concentration, add 5mg / ml MTT to the cell culture wells after 72 hours of culture, 10ul per well, and place at 37°C After incubation for 3 hours, 150ul of DMSO was added to completely dissolve the formazan, and a microplate reader was used for colorimetry at a wavelength of 570nm. Calculate the median toxic concentration of...
Embodiment 2
[0045] Example 2 Effect of isochlorogenic acid composition on liver cell injury induced by D-GalN (galactosamine)
[0046] Human normal liver cell line HL-7702, placed at 37°C in CO 2 Culture in an incubator; after the hepatocytes adhered to the wall, replace the culture medium, add 20mmol / l D-GalN, and act for 8h; then discard the culture medium, and add 200μl of different concentrations of the test drug solution, each concentration was set at 6 Repeat wells, set solvent and positive control at the same time; continue to culture for 48h, add 10μl MTT (5mg / ml) to each well, act for 4h, aspirate and discard the supernatant, add 150μl DMSO, shake and mix well; The instrument measures the A value. Protection evaluation: calculation of protection rate (%) and proliferation index.
[0047]
[0048] Note 2: Proliferation index = sample group (A) / injury group (A)
[0049] Table 2 Effect of isochlorogenic acid composition on D-GalN-induced normal human liver cell damage
[0050...
Embodiment 3
[0053] Example 3 Protective effect of isochlorogenic acid composition on D-GalN-induced acute liver injury model in mice
[0054] Materials: 20-25g male ICR mice were randomly divided into groups according to body weight, 10 mice in each group. A normal control group, an injury control group, a positive control group and high, medium and low dose groups of the test drug were set up. Animals in the drug group were given prophylactic administration once a day for 7 consecutive days. The normal control group and the injury control group were given an equal volume of solvent, and the positive control group was given 50 mg / kg of silymarin. One hour after the last administration, animals in each group except the normal group were injected with 650 mg / kg galactosamine (D-GalN) intraperitoneally to cause toxicity. 24 hours after the last administration, the serum was separated to measure AST and ALT. At the same time, liver tissue was taken for making pathological sections.
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