| In recent years,toxicity reported for traditional Chinese medicine and its preparation is increasing,traditional Chinese medjcine-induced liver injury,is one of the four common causes of drug-induced liver injury.objective:To observe the traditional Chinese medicine actiVe anticancer ingredient djsodium cantharidinate,gallic acid,ursolic acid on normal human hepatocytes LO2.provide clues to find efffectiVe attenuated efficiency antagonist..Methods:MTT assay of these three anticancer constltuents of cell proliferation inhibition. inverted microscope, fluorescent staining and electron microscopy to determine the morphological changes of cells,Giemsa staining to detect micro-nuclear rate,flow cytometry to measure the apoptosis rate,Western blot to determine the expression changes of proteins within the cells of each administration group.Results:Found0.625μg/ml group1.25μg/ml group,the the2.5μg/ml grouUp cantharides of sodium for24hours,inhibition of LO2cells were9.1%,17.1%,23.9%；SMMC-7721,The inhibition rates were11.2%,23.5%,30.7%,respectiVely.No significant change in the nuorescence microscope0.625μg/ml group of1.25,2.5μg/ml group LO2cells chromosome breakage；Giemsa staining under light microscope counting0.625μg/ml group no significant changes in1.25μg/ml group of micro-nuclear rate of46.0‰,52.2‰2.5μg/ml group；0.625μg/ml apoptotic rate of5.08%and16-31%in1.25μg/ml the2.5μg/ml group21.86%,a dose dependent manner；electron microscopy confirmed that the2.5μg/ml group,cell shrinkage, surface microvilli,mitochondrial swelling,spinal fracture,intracellular vacuoles increased nuclei condensation and fragmentation,characterislic change was apoptosis.1.25ug/ml and2.5ug/ml groups of Caspase-3,Bcl-2content decreased,Bax increased with the negatiVe contro1group was statistically significant(P<0.05),NFkb increase slightly,with the negative contro1group Compared to no difference.Ursolic acid for24h,the inhibitory rate of12.5μmol/L group,25μmol/L,50μmol/L group LO2cells were5.4%,21.4%,49.8%inhibition of SMMC-7721rate of17.6%,respectively,36.1%,60.9%.50μmol/L group,nuclear condensation under a fluorescence microscope,the brightness increased,the formation of crescent-shaped,bias cell side,and even cell debris and apoptotic bodies.Streaming results showed that the apoptosis rate of48.76%.Gallic acid for24h,each dose group showed LO2and SMMC-7721cell proliferation inhibition,but affer72h,the inhibition of proliferation of SMMC-7721cells than LO2celJ proliferation inhibition.Gallic acid for24h,100μmol/L,group of cells appears obviOUS morphological changes of cell apoptosis rate was12.53%,elevated Caspase-3and P53 expression.Conclusions:The cantharides sodium, ursolic acid and gallic acid on hepatoma cells and normal liver cells was inhibited. The cantharides sodium inhibition of normal liver cells is lower than the inhibition of hepatoma cells; in dose can cause inhibition of proliferation of normal liver cells, chromosome damage and cell apoptosis; the cantharides sodium caused cell apoptosis, which may and Caspase-3, Bcl-2and Bax protein expression related to the change. Ursolic acid inhibition of the normal liver cells is lower than the inhibition of hepatoma cells; ursolic acid can be caused by the normal liver cell proliferation inhibition at25μmol/L,50μmol/L not only inhibit the proliferation of liver cells, can also lead cells apoptosis; ursolic acid-induced apoptosis, Caspase-3protein expression changes related. Gallic acid administered administered72h after the inhibition of liver cancer cells more than the suppression of the normal liver cells; gallic acid100μmol/L, caused by human normal liver cell apoptosis, consider changes in P53and Caspase-3protein. |
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